**Chapter 2** Applications of Oxidoreductases

*Sandhya Rani Gogoi*

### **Abstract**

phenolic extract of jaboticaba (plinia peruviana). Journal of Food Science and technology. 2019;**56**(3):1165-1173

An unusual cause of recurrent lactic acidosis in a paediatric critical care unit. Journal of Critical Care Medicine. 2019;

[60] Dean B, Chrisp GL, Quartararo M, et al. P450 oxidoreductase deficiency: A systematic review and meta-analysis of genotypes, phenotypes, and their relationships. The Journal of Clinical Endocrinology and Metabolism. 2020;

[61] Cheng JB et al. Molecular genetics of 3beta-hydroxy-delta5-C27-steroid oxidoreductase deficiency in 16 patients with loss of bile acid synthesis and liver

disease. The Journal of Clinical Endocrinology and Metabolism. 2003;

prenylation and therefore is

Disease. 2019;**10**(2):91

[62] Lacher S, Bruttger J, Kalt B, et al. HMG-CoA reductase promotes protein

indispensible for T-cell survival. Cell Death & Disease. 2017;**8**:e2824

[63] Göbel A, Breining D, Rauner M, Hofbauer LC, Rachner TD. Induction of 3-hydroxy-3-methylglutaryl-CoA reductase mediates statin resistance in breast cancer cells. Cell Death &

**5**(2):71-75

**105**(3):e42-e52

**88**(4):1833-1841

[52] Pandey VP, Awasthi M, Singh S, Tiwari S, Dwivedi UN. Comprehensive review on function and application of

[53] Garrone A, Archipowa N, Zipfel PF,

Protochlorophyllide Oxidoreductases A and B – Catalytic Efficiency and Initial Reaction Steps. Journal of biological chemistry. 2015;**290**(47):28530-28539

[54] Ho T, Chang C, Wu J, Huang I, et al. Recombinant expression of aldehyde dehydrogenase 2 (ALDH2) in *Escherichia coli* nissle 1917 for oral delivery in ALDH2-deficient individuals. bioRxiv. 2019. preprint

[55] Shi Y, van Rhijn JR, Bormann M, Mossink B, et al. Brunner syndrome associated MAOA dysfunction in human induced dopaminergic neurons results in dysregulated NMDAR expression and increased network activity. bioRxiv.

[56] Nsiah-Sefaa A, McKenzie M. Combined defects in oxidative phosphorylation and fatty acid βoxidation in mitochondrial disease. Bioscience Reports. 2016;**36** art: e00313

[57] Chen X, Qi F, Dash RK, Beard DA. Kinetics and regulation of mammalian NADH-ubiquinone oxidoreductase (Complex I). Biophysical Journal. 2010;

[58] Scheffler IE. Mitochondrial disease associated with complex I (NADH-CoQ oxidoreductase) deficiency. Journal of Inherited Metabolic Disease. 2015;**38**(3):

[59] Gupta N, Rutledge C. Pyruvate dehydrogenase complex deficiency:

2019. preprint

**99**:1426-1436

405-415

**16**

plant peroxidases. Journal of Biochemistry & Analytical Biochemistry. 2017;**6**:308

*Oxidoreductase*

Hermann G, Dietzek B. Plant

Oxidoreductases comprise of a large group of enzymes catalyzing the transfer of electrons from an electron donor to an electron acceptor molecule, commonly taking nicotinamide adenine dinucleotide phosphate (NADP) or nicotinamide adenine dinucleotide (NAD) as cofactors. Research on the potential applications of oxidoreductases on the growth of oxidoreductase-based diagnostic tests and better biosensors, in the design of inventive systems for crucial coenzymes regeneration, and in the creation of oxidoreductase-based approaches for synthesis of polymers and oxyfunctionalized organic substrates have made great progress. This chapter focuses on biocatalytic applications of oxidoreductases, since many chemical and biochemical transformations involve oxidation/reduction processes, developing practical applications of oxidoreductases has long been a significant target in biotechnology. Oxidoreductases are appropriate catalysts owing to their biodegradability, specificity and efficiency and may be employed as improved biocatalysts to substitute the toxic/expensive chemicals, save on energy/resources consumption, generate novel functionalities, or reduce complicated impacts on environment.

**Keywords:** oxidoreductases, cofactors, biosensors, coenzymes regeneration, biocatalytic

#### **1. Introduction**

The various chemical transformations catalyzed by enzymes make these catalysts a key goal for utilization by the promising biotechnology industries. In the recent years, intense research in the field of enzyme technology has provided numerous approaches that facilitate the practical application of enzymes. This chapter emphasizes the application of oxidoreductases which catalyze the exchange of electrons amid the donor and acceptor molecules, in reactions involving electron transfer, proton/hydrogen extraction, hydride transfer, oxygen insertion, or other imperative steps. Oxidoreductases acquire advantage from the inclusion of different cofactors - for instance heme, flavin and metal ions - to catalyze redox reactions [1]. Majority of oxidoreductases are nicotinamide cofactor-dependent enzymes which have a high preference for nicotinamide adenine dinucleotide phosphate (NADP) or nicotinamide adenine dinucleotide (NAD) and they are further classified in six major classes which are oxidases, dehydrogenases, hydroxylases, oxygenases, peroxidases and reductases [2]. This chapter demonstrates the potential applications of oxidoreductases on the growth of oxidoreductase-based diagnostic tests and better biosensors, in the design of inventive systems for crucial coenzymes regeneration, and in the formation of oxidoreductase-based approaches for synthesis of polymers and oxyfunctionalized organic substrates.

#### **2. Oxidoreductase-based diagnostic tests and as biosensors**

The diagnosis and monitoring of a variety of diseases is extremely demanding nowadays for routine examination of clinical samples and other associated tests. The diagnostic enzymes are used for the detection/diagnosis or prognosis of disease conditions due to their substrate specificity and quantitated activity in the presence of other proteins, and are preferred in diagnosis, which can be used as a diagnostic tool for disease detection [3]. Depending on the verity of the disease, diseased state often leads to tissue damage. In such conditions, enzymes specific to diseased organs are released into blood circulation with augmented enzyme activity. The measurement of corresponding enzyme activities in blood/plasma, or any other body fluid, has been exploited in the diagnosis of diseased tissues/organs [3].

Jixu Wang et al. [4] investigated the expression and significance of glucose-6-phosphate dehydrogenase (G6PD) in human gastric cancer progression and prognosis. Apoptosis and necrosis are two major types of cell death in normal and disease pathologies. A key signature for necrotic cells is the permeabilization of the plasma membrane which can be quantified in tissue culture settings by measuring the release of the intracellular enzyme lactate dehydrogenase (LDH). It has been described that the measuring LDH release is a useful method for the detection of necrosis [5]. Two dehydrogenases, specifically, sorbitol dehydrogenase (SDH) and LDH, are used for cancer prognosis [3]. Reports suggested that in prostate cancer [6], and precancerous colorectal neoplasms [7], an abnormal serum concentration of SDH has been observed. Additionally, an enhanced level of SDH can be observed in acute liver damage and parenchymal hepaticdiseases [3]. It has been reported that LDH, marker of anaerobic metabolism, is associated with highly invasive and metastatic breast cancer and suggested that the association of activity of LDH in tumor tissue with mammographic characteristics could help in defining aggressive breast cancers [8]. The gene expression of LDH is studied in several human malignant tumors, collectively among colorectal cancer [9], lung cancer [10–12], breast cancer [13], oral cancer [14], prostate cancer [15], germ cell cancer [16], and pancreatic cancer [17]. In recent times, the prognostic value of the serum LDH level in cancer patients has been considered as a significant area of research. Additionally, LDH performs as a prognostic marker in patients with acute leukemia [18] and sickle cell disease [19].

A biosensor is an analytical tool that comprises a biological or biologically derived sensing matter with close proximity to the physico-chemical transducer [3]. The chief function of such a device is to produce a discrete or uninterrupted signal that is comparative to the concentration of the analyte [20]. Enzyme-based chemical biosensors are based on biological recognition and in order to function, the enzymes must be accessible to catalyze a specific biochemical reaction and be stable under the normal operating circumstances of the biosensor [21]. Generally the function of oxidoreductase biosensors is dependent on charge transport amid the enzyme and an electrode surface by means of coenzymes or redox mediators [22].

Over the years, various enzyme-based biosensors have been developed, however only a few of them are commercialized. The majority of the published work on enzymatic biosensors focuses on targeted blood glucose monitoring based on amperometric techniques [3]. The earliest glucose biosensor based on glucose dehydrogenase from Erwinia sp. and carbon paste was generated by Laurinavicius et al. [23] where the enzyme was incorporated in a polylysine-albumin gel, and the anchoring material was a paste of chemically adapted carbon powder, fumed silica, and binding material. A cellulose dehydrogenase based glucose biosensor from a mutant of *Corynascus thermophilus* has been developed, and a glassy carbon electrode (GCE) was acquired

**19**

*Applications of Oxidoreductases*

Glucose oxidase

Oxalate oxidase

Lactate oxidase

**Table 1.**

Cholesterol oxidase

*DOI: http://dx.doi.org/10.5772/intechopen.94409*

Glucose Blood plasma,

Oxalate Blood serum

Lactate Blood plasma,

blood serum, urine, and saliva

and urine

blood serum, drug and biological samples

"Biostator" by Miles (Elkhart, Indiana).

*Oxidoreductase enzymatic biosensors as diagnostic tools.*

**3. Oxidoreductases in coenzymes regeneration**

10% of them require NADP as a coenzyme [44].

by direct electrode position of gold nanoparticles (AuNPs). The biosensor was used for the detection of glucose in human saliva samples, with successful results in terms of both revival and association with glucose blood levels [24]. This proposes the development of noninvasive glucose monitoring devices. The details of different oxidoreductase enzymatic biosensors applied for clinical diagnosis are listed in **Table 1**. The first marketable biosensor (glucose biosensor) was commenced in 1975 which was derived from the electrochemical recognition of hydrogen peroxide, and the glucose oxidase was employed for the improvement of the biosensor [3]. Subsequently, Clemens et al. [25] established a novel amperometric glucose biosensor in a bedside artificial pancreas, and it was marked underneath the brand name

**Enzymes Analyte Test sample Disease diagnosed References**

Cholesterol Blood serum Coronary heart disease, myocardial

Diabetes, hypoglycemia [26–29]

[30]

[31–34]

[35–40]

Idiopathic urolithiasis and various

and cerebral infarction (stroke)

Hyper lactatemia, cardiac arrest, resuscitation, sepsis, reduced renal excretion, decreased extra hepatic metabolism, intestinal infarction

intestinal diseases

and lacticacidosis

The most of oxidoreductases for catabolism and anabolism significantly require two natural nicotinamide-based coenzymes (NAD and NADP), respectively. The most NAD(P)-dependent oxidoreductases choose one coenzyme as an electron acceptor or donor to the other depending on their diverse metabolic functions [41]. Generally coenzymes are involved in these oxidoreductase-catalyzed reactions to transport electron, hydride, hydrogen, oxygen, or other atoms or small molecules in diverse enzymatic pathways [42, 43]. The nicotinamide adenine dinucleotide (NAD)/nicotinamide adenine dinucleotide phosphate (NADP), ubiquinone (CoQ ), and flavin mononucleotide (FMN)/flavin adenine dinucleotide (FAD) are the typical coenzymes. Nicotinamide-based coenzymes for the electron transport and storage in the form of hydride groups are the most noteworthy in view of the fact that 80% of characterized oxidoreductases necessitate NAD as a coenzyme, and

Nicotinamide coenzymes based dehydrogenases are of emergent importance for the production of chiral compounds, either by reduction of a prochiral precursor or via oxidative resolution of their racemate [45]. Nevertheless, the oxidized and reduced nicotinamide cofactors regeneration is an extremely critical step as the employ of these cofactors in stoichiometric amounts is too expensive for function. There are very few enzymes which are appropriate for the regeneration of oxidized


**Table 1.**

*Oxidoreductase*

**2. Oxidoreductase-based diagnostic tests and as biosensors**

The diagnosis and monitoring of a variety of diseases is extremely demanding nowadays for routine examination of clinical samples and other associated tests. The diagnostic enzymes are used for the detection/diagnosis or prognosis of disease conditions due to their substrate specificity and quantitated activity in the presence of other proteins, and are preferred in diagnosis, which can be used as a diagnostic tool for disease detection [3]. Depending on the verity of the disease, diseased state often leads to tissue damage. In such conditions, enzymes specific to diseased organs are released into blood circulation with augmented enzyme activity. The measurement of corresponding enzyme activities in blood/plasma, or any other body fluid, has been exploited in the diagnosis of diseased tissues/organs [3]. Jixu Wang et al. [4] investigated the expression and significance of glucose-6-phosphate dehydrogenase (G6PD) in human gastric cancer progression and prognosis. Apoptosis and necrosis are two major types of cell death in normal and disease pathologies. A key signature for necrotic cells is the permeabilization of the plasma membrane which can be quantified in tissue culture settings by measuring the release of the intracellular enzyme lactate dehydrogenase (LDH). It has been described that the measuring LDH release is a useful method for the detection of necrosis [5]. Two dehydrogenases, specifically, sorbitol dehydrogenase (SDH) and LDH, are used for cancer prognosis [3]. Reports suggested that in prostate cancer [6], and precancerous colorectal neoplasms [7], an abnormal serum concentration of SDH has been observed. Additionally, an enhanced level of SDH can be observed in acute liver damage and parenchymal hepaticdiseases [3]. It has been reported that LDH, marker of anaerobic metabolism, is associated with highly invasive and metastatic breast cancer and suggested that the association of activity of LDH in tumor tissue with mammographic characteristics could help in defining aggressive breast cancers [8]. The gene expression of LDH is studied in several human malignant tumors, collectively among colorectal cancer [9], lung cancer [10–12], breast cancer [13], oral cancer [14], prostate cancer [15], germ cell cancer [16], and pancreatic cancer [17]. In recent times, the prognostic value of the serum LDH level in cancer patients has been considered as a significant area of research. Additionally, LDH performs as a prognostic marker in patients with acute leukemia

A biosensor is an analytical tool that comprises a biological or biologically derived sensing matter with close proximity to the physico-chemical transducer [3]. The chief function of such a device is to produce a discrete or uninterrupted signal that is comparative to the concentration of the analyte [20]. Enzyme-based chemical biosensors are based on biological recognition and in order to function, the enzymes must be accessible to catalyze a specific biochemical reaction and be stable under the normal operating circumstances of the biosensor [21]. Generally the function of oxidoreductase biosensors is dependent on charge transport amid the enzyme and

Over the years, various enzyme-based biosensors have been developed, however only a few of them are commercialized. The majority of the published work on enzymatic biosensors focuses on targeted blood glucose monitoring based on amperometric techniques [3]. The earliest glucose biosensor based on glucose dehydrogenase from Erwinia sp. and carbon paste was generated by Laurinavicius et al. [23] where the enzyme was incorporated in a polylysine-albumin gel, and the anchoring material was a paste of chemically adapted carbon powder, fumed silica, and binding material. A cellulose dehydrogenase based glucose biosensor from a mutant of *Corynascus thermophilus* has been developed, and a glassy carbon electrode (GCE) was acquired

an electrode surface by means of coenzymes or redox mediators [22].

**18**

[18] and sickle cell disease [19].

*Oxidoreductase enzymatic biosensors as diagnostic tools.*

by direct electrode position of gold nanoparticles (AuNPs). The biosensor was used for the detection of glucose in human saliva samples, with successful results in terms of both revival and association with glucose blood levels [24]. This proposes the development of noninvasive glucose monitoring devices. The details of different oxidoreductase enzymatic biosensors applied for clinical diagnosis are listed in **Table 1**. The first marketable biosensor (glucose biosensor) was commenced in 1975 which was derived from the electrochemical recognition of hydrogen peroxide, and the glucose oxidase was employed for the improvement of the biosensor [3]. Subsequently, Clemens et al. [25] established a novel amperometric glucose biosensor in a bedside artificial pancreas, and it was marked underneath the brand name "Biostator" by Miles (Elkhart, Indiana).

#### **3. Oxidoreductases in coenzymes regeneration**

The most of oxidoreductases for catabolism and anabolism significantly require two natural nicotinamide-based coenzymes (NAD and NADP), respectively. The most NAD(P)-dependent oxidoreductases choose one coenzyme as an electron acceptor or donor to the other depending on their diverse metabolic functions [41]. Generally coenzymes are involved in these oxidoreductase-catalyzed reactions to transport electron, hydride, hydrogen, oxygen, or other atoms or small molecules in diverse enzymatic pathways [42, 43]. The nicotinamide adenine dinucleotide (NAD)/nicotinamide adenine dinucleotide phosphate (NADP), ubiquinone (CoQ ), and flavin mononucleotide (FMN)/flavin adenine dinucleotide (FAD) are the typical coenzymes. Nicotinamide-based coenzymes for the electron transport and storage in the form of hydride groups are the most noteworthy in view of the fact that 80% of characterized oxidoreductases necessitate NAD as a coenzyme, and 10% of them require NADP as a coenzyme [44].

Nicotinamide coenzymes based dehydrogenases are of emergent importance for the production of chiral compounds, either by reduction of a prochiral precursor or via oxidative resolution of their racemate [45]. Nevertheless, the oxidized and reduced nicotinamide cofactors regeneration is an extremely critical step as the employ of these cofactors in stoichiometric amounts is too expensive for function. There are very few enzymes which are appropriate for the regeneration of oxidized

nicotinamide cofactors. Glutamate dehydrogenase can be utilized for the oxidation of NADH in addition to NADPH while l-lactate dehydrogenase is able to oxidize NADH only [45]. The reduction of NAD<sup>+</sup> is carried out by formate and FDH [45]. Glucose-6-phosphate dehydrogenase and glucose dehydrogenase are proficient to reduce both NAD+ and NADP+ [45]. It has been reported that ADH from horse liver reduces NAD+ whereas ADHs from *Lactobacillus* strains catalyze the reduction of NADP+ [45]. These enzymes can be applied by their inclusion in entire cell biotransformations by an NAD(P)+ -dependent major reaction to achieve *in situ* regeneration of the consumed cofactor [45]. And for the regeneration of the reduced cofactors NADH and NADPH numerous systems for instance engineered formate dehydrogenase [46, 47], phosphite dehydrogenase [48, 49], glucose dehydrogenase [50, 51] plus cosubstrate are well established and extensively used.

Johannes et al. [52] reported the engineering of a highly stable and active mutant phosphite dehydrogenase (12x-A176R PTDH) from *Pseudomonas stutzeri* and evaluation of its potential as an effective NADPH regeneration system in an enzyme membrane reactor. They have utilized two practically imperative enzymatic reactions including xylose reductase-catalyzed xylitol synthesis and alcohol dehydrogenase-catalyzed (R)-phenylethanol synthesis as models, and the mutant PTDH was compared to the commercially available NADP+ -specific *Pseudomonas sp*. 101 formate dehydrogenase (mut Pse-FDH) that is extensively employed for NADPH regeneration [52]. Soluble water-forming NAD(P)H oxidases comprise a promising NAD(P)+ regeneration scheme since they only require oxygen as cosubstrate and produce water as only byproduct [53]. In addition, the thermodynamic equilibrium of O2 reduction is a significant driving force for mostly energetically unfavorable biocatalytic oxidations [53]. Petschacher et al. [53] presented the generation of an NAD(P)H oxidase with high activity for both cofactors, NADH and NADPH. Applicability for cofactor regeneration is shown for coupling with alcohol dehydrogenase from *Sphyngobium yanoikuyae* for 2-heptanone production.

#### **4. Oxidoreductase-based approaches for synthesis of polymers and various organic substrates**

Enzyme catalyzed oxidation reactions have achieved growing concern in biocatalysis recently, reflected also by numerous outstanding reviews on this topic reported in the last years [54–56]. The group of oxidoreductases, to which all enzyme catalyzing oxidoreduction reactions, comprises numerous groups of biocatalysts such as dehydrogenases, monooxygenases, dioxygenases, oxidases, peroxidases, etc. [55]. Moreover, the enzymatic oxidative polymerizations have advantages of using nontoxic catalysts and mild reaction conditions, and the specific enzyme catalysis affords regio- and chemoselective polymerizations to construct functional materials [57]. It has been reported that peroxidases with the use of hydrogen peroxide as oxidant efficiently induce the oxidative coupling of phenols to phenolic polymers, the majority of which are scarcely attained by conventional chemical catalysts [57]. In addition, it has been published that laccase and peroxidase are helpful for production of cross-linked polymers such as artificial urushi and biopolymer hydrogel [57]. Kobayashi [58] established that the enzymatic polymerization as to be an efficient method of polymer synthesis. The polymerization uses hydrolases and oxidoreductases as catalysts and this new method of polymer synthesis afforded natural polysaccharides like cellulose, amylose, xylan, and chitin, and unnatural polysaccharides catalyzed by a glycosidase from welldesigned monomers, varied functionalized polyesters catalyzed by lipase from a variety of monomers, and poly-aromatics materials catalyzed by an oxidoreductase

**21**

*Applications of Oxidoreductases*

photobiocatalytic redox reactions.

**5. Medical applications**

*DOI: http://dx.doi.org/10.5772/intechopen.94409*

polymerization has been initiated by oxidoreductase [58].

and an enzyme model complex from phenols and anilines [58]. Furthermore, vinyl

Marjanovic et al. [59] reviewed the oxidative oligomerization and polymerization of various arylamines, e.g., aniline, substituted anilines, aminonaphthalene and its derivatives, catalyzed by oxidoreductases, such as laccases and peroxidases, in aqueous, organic, and mixed aqueous organic monophasic or biphasic media. Owing to the nontoxicity of oxidoreductases and their elevated catalytic effectiveness, as well as high selectivity of enzymatic oligomerizations/polymerizations under gentle conditions by means of primarily water as a solvent and often resulting in minimal byproduct formation enzymatic oligomerizations and polymerizations of arylamines are environmentally friendly and considerably contribute to a "green"

It has been also established that oxidative enzymes comprise privileged catalysts in organic synthesis [60]. Environmentally benign reaction conditions with high selectivity are the most fascinating characteristic exhibited by these biocatalysts in contrast to classical metal-based reagents. de Gonzalo et al. [60] reviewed the new perspectives and concepts derived from oxidative enzymatic processes, involving oxidative C-C bond forming reactions, atroposelective oxidations, oxidative dynamic processes, interconnected reactions, cyclic deracemizations, oxidative desymmetrizations and artificial oxidative enzymes. Oxidoreductases comprise an imperative group of biocatalysts as they facilitate not merely the broadly used stereoselective reduction of aldehydes and ketones but also the less well exploited oxidation of alcohols and amines [53]. In addition, oxidoreductases catalyzed oxidations are utilized for production of chiral alcohols and amines by deracemization [54, 60–62]. It has been reviewed thoroughly that the oxidoreductases enable chemists to perform highly selective and efficient transformations ranging from simple alcohol oxidations to stereoselective halogenations of non-activated C-H bonds [63]. Mifsud et al. [64] demonstrated for the first time that catalytic water oxidation mediated by robust TiO2 semiconductors can be productively coupled to oxidoreductases achieving

One of the major applications of oxidoreductase is a pharmaceutical synthesis of 3,4-dihydroxylphenyl alanine (DOPA), which is employed in the treatment of Parkinson's disease and the industrial process that synthesizes DOPA make use of the oxidoreductase polyphenol oxidase [65]. It has been reported that the enantioselective reduction of C-4-substituted 3,5-dixocarboxylates can be carried out by using alcohol dehydrogenase from *Lactobacillus brevis* (LBADH) over-expressed in *E. coli* [66]. Laccase can be employed to synthesize numerous complex medicinal agents including triazolo(benzo)cycloalkyl thiadiazines, vinblastine, penicillin X dimer, cephalosporin antibiotics, and dimerized vindo-line [67]. In addition laccase can be used to synthesize a range of functional organic compounds including polymers with specific mechanical/electrical/optical properties, textile dyes, cosmetic pigments, flavor agents, and pesticides [68]. Biocatalysis is facilitating technology to organic synthesis chemistry by providing high selectivity of enzymatic reactions

under mild conditions makes it a very valuable tool for green chemistry.

Due to the specificity and bio-based nature, potential applications of oxidoreductases in various fields are attracting active research efforts [69]. Several products generated by oxidoreductases are finding applications as antimicrobial, detoxifying, or active personal-care agents [69]. One potential application is laccase-based *in situ* generation of iodine, a reagent extensively used as disinfectant [67]. It has been

chemistry of conducting and redox-active oligomers and polymers [59].

#### *Applications of Oxidoreductases DOI: http://dx.doi.org/10.5772/intechopen.94409*

*Oxidoreductase*

reduce both NAD+

formations by an NAD(P)+

reduces NAD+

NADP+

NAD(P)+

NADH only [45]. The reduction of NAD<sup>+</sup>

and NADP+

plus cosubstrate are well established and extensively used.

was compared to the commercially available NADP+

**and various organic substrates**

genase from *Sphyngobium yanoikuyae* for 2-heptanone production.

**4. Oxidoreductase-based approaches for synthesis of polymers** 

Enzyme catalyzed oxidation reactions have achieved growing concern in biocatalysis recently, reflected also by numerous outstanding reviews on this topic reported in the last years [54–56]. The group of oxidoreductases, to which all enzyme catalyzing oxidoreduction reactions, comprises numerous groups of biocatalysts such as dehydrogenases, monooxygenases, dioxygenases, oxidases, peroxidases, etc. [55]. Moreover, the enzymatic oxidative polymerizations have advantages of using nontoxic catalysts and mild reaction conditions, and the specific enzyme catalysis affords regio- and chemoselective polymerizations to construct functional materials [57]. It has been reported that peroxidases with the use of hydrogen peroxide as oxidant efficiently induce the oxidative coupling of phenols to phenolic polymers, the majority of which are scarcely attained by conventional chemical catalysts [57]. In addition, it has been published that laccase and peroxidase are helpful for production of cross-linked polymers such as artificial urushi and biopolymer hydrogel [57]. Kobayashi [58] established that the enzymatic polymerization as to be an efficient method of polymer synthesis. The polymerization uses hydrolases and oxidoreductases as catalysts and this new method of polymer synthesis afforded natural polysaccharides like cellulose, amylose, xylan, and chitin, and unnatural polysaccharides catalyzed by a glycosidase from welldesigned monomers, varied functionalized polyesters catalyzed by lipase from a variety of monomers, and poly-aromatics materials catalyzed by an oxidoreductase

nicotinamide cofactors. Glutamate dehydrogenase can be utilized for the oxidation of NADH in addition to NADPH while l-lactate dehydrogenase is able to oxidize

Glucose-6-phosphate dehydrogenase and glucose dehydrogenase are proficient to

of the consumed cofactor [45]. And for the regeneration of the reduced cofactors NADH and NADPH numerous systems for instance engineered formate dehydrogenase [46, 47], phosphite dehydrogenase [48, 49], glucose dehydrogenase [50, 51]

phosphite dehydrogenase (12x-A176R PTDH) from *Pseudomonas stutzeri* and evaluation of its potential as an effective NADPH regeneration system in an enzyme membrane reactor. They have utilized two practically imperative enzymatic reactions including xylose reductase-catalyzed xylitol synthesis and alcohol dehydrogenase-catalyzed (R)-phenylethanol synthesis as models, and the mutant PTDH

formate dehydrogenase (mut Pse-FDH) that is extensively employed for NADPH regeneration [52]. Soluble water-forming NAD(P)H oxidases comprise a promising

 regeneration scheme since they only require oxygen as cosubstrate and produce water as only byproduct [53]. In addition, the thermodynamic equilibrium of O2 reduction is a significant driving force for mostly energetically unfavorable biocatalytic oxidations [53]. Petschacher et al. [53] presented the generation of an NAD(P)H oxidase with high activity for both cofactors, NADH and NADPH. Applicability for cofactor regeneration is shown for coupling with alcohol dehydro-

whereas ADHs from *Lactobacillus* strains catalyze the reduction of

[45]. These enzymes can be applied by their inclusion in entire cell biotrans-

Johannes et al. [52] reported the engineering of a highly stable and active mutant

is carried out by formate and FDH [45].


[45]. It has been reported that ADH from horse liver


**20**

and an enzyme model complex from phenols and anilines [58]. Furthermore, vinyl polymerization has been initiated by oxidoreductase [58].

Marjanovic et al. [59] reviewed the oxidative oligomerization and polymerization of various arylamines, e.g., aniline, substituted anilines, aminonaphthalene and its derivatives, catalyzed by oxidoreductases, such as laccases and peroxidases, in aqueous, organic, and mixed aqueous organic monophasic or biphasic media. Owing to the nontoxicity of oxidoreductases and their elevated catalytic effectiveness, as well as high selectivity of enzymatic oligomerizations/polymerizations under gentle conditions by means of primarily water as a solvent and often resulting in minimal byproduct formation enzymatic oligomerizations and polymerizations of arylamines are environmentally friendly and considerably contribute to a "green" chemistry of conducting and redox-active oligomers and polymers [59].

It has been also established that oxidative enzymes comprise privileged catalysts in organic synthesis [60]. Environmentally benign reaction conditions with high selectivity are the most fascinating characteristic exhibited by these biocatalysts in contrast to classical metal-based reagents. de Gonzalo et al. [60] reviewed the new perspectives and concepts derived from oxidative enzymatic processes, involving oxidative C-C bond forming reactions, atroposelective oxidations, oxidative dynamic processes, interconnected reactions, cyclic deracemizations, oxidative desymmetrizations and artificial oxidative enzymes. Oxidoreductases comprise an imperative group of biocatalysts as they facilitate not merely the broadly used stereoselective reduction of aldehydes and ketones but also the less well exploited oxidation of alcohols and amines [53]. In addition, oxidoreductases catalyzed oxidations are utilized for production of chiral alcohols and amines by deracemization [54, 60–62]. It has been reviewed thoroughly that the oxidoreductases enable chemists to perform highly selective and efficient transformations ranging from simple alcohol oxidations to stereoselective halogenations of non-activated C-H bonds [63]. Mifsud et al. [64] demonstrated for the first time that catalytic water oxidation mediated by robust TiO2 semiconductors can be productively coupled to oxidoreductases achieving photobiocatalytic redox reactions.

One of the major applications of oxidoreductase is a pharmaceutical synthesis of 3,4-dihydroxylphenyl alanine (DOPA), which is employed in the treatment of Parkinson's disease and the industrial process that synthesizes DOPA make use of the oxidoreductase polyphenol oxidase [65]. It has been reported that the enantioselective reduction of C-4-substituted 3,5-dixocarboxylates can be carried out by using alcohol dehydrogenase from *Lactobacillus brevis* (LBADH) over-expressed in *E. coli* [66]. Laccase can be employed to synthesize numerous complex medicinal agents including triazolo(benzo)cycloalkyl thiadiazines, vinblastine, penicillin X dimer, cephalosporin antibiotics, and dimerized vindo-line [67]. In addition laccase can be used to synthesize a range of functional organic compounds including polymers with specific mechanical/electrical/optical properties, textile dyes, cosmetic pigments, flavor agents, and pesticides [68]. Biocatalysis is facilitating technology to organic synthesis chemistry by providing high selectivity of enzymatic reactions under mild conditions makes it a very valuable tool for green chemistry.

#### **5. Medical applications**

Due to the specificity and bio-based nature, potential applications of oxidoreductases in various fields are attracting active research efforts [69]. Several products generated by oxidoreductases are finding applications as antimicrobial, detoxifying, or active personal-care agents [69]. One potential application is laccase-based *in situ* generation of iodine, a reagent extensively used as disinfectant [67]. It has been

described that laccase-iodide salt binary iodine-generating system (for sterilization) can have several advantages over the direct iodine application [69]. Peroxidases may replace laccase for the application, even though they would require H2O2 as cosubstrate [69]. The ClO¯ and Mn(III) species formed by haloperoxidase and Mn-peroxidase are extremely effective oxidants and antimicrobial agents [70]. Peroxidase can also be used to cross-link collagen which is beneficial to the healing of damaged skin [71]. The physiological activities of lysyl oxidase comprise the extracellular matrix construction which can hasten wound-healing [72, 73]. A glucose oxidase, lactoperoxidase, and iodide system has been tested for dental care and the oxidase produces H2O2 to feed the peroxidase, so that it can produce iodine that can kill plaque-causing bacteria [74]. It has been reported that the haloperoxidase can be used to oxidatively modify rubber latex surfaces, making them less allergenic [75]. A secreted oxidoreductase may even be developed as a vaccine against secretor microbes such as, *Aspergillus oryzae* catalase A protein has been studied as a potential aspergillosis vaccine [69]. It has been reported that low-molecular-mass laccase purified from the mushroom *Tricholoma giganteumis* possesses significant HIV-1 reverse transcriptase inhibitory activity [76]. As nature's own catalysts, enzymes acquire very diverse specificity, reactivity, and other physicochemical, catalytic, and biological properties highly enviable for miscellaneous industrial and medical applications [69].

#### **6. Conclusions**

Tremendous progress has been made in the recent years in the field of applications of oxidoreductases. Oxidoreductases metabolism is a fundamental bioprocess that plays a pivotal role in all species, including humans, plants, animals, and microorganisms, as their specific function is to catalyze oxidation and reduction reactions that occur within the cell. Abnormality in this metabolic system leads to a number of metabolic disorders. Thus, owing to the remarkable properties of oxidoreductases, they can be used for the diagnosis of disorders. They can provide insight into the diseased state by diagnosis, prognosis, or by assessment of response therapy. It has been established that oxidoreductases as biosensors are becoming popular potential tools in biotechnology due to their high specificity. With oxidoreductases, the conversion of a variety of aliphatic/aromatic molecules can be achieved; inert hydrocarbons can be functionalized (by hydroxylation, sulfoxidation, epoxidation, etc.); regio-, enantio- (on racemic substrates); enantiotopo– (on prochiral sub-strates); and chemo-selective reactions can be accomplished; important synthons from inexpensive and renewable biomaterials can be constructed; and the negative environment impact can be reduced [69]. Since numerous chemical and biochemical transformations engage oxidation/reduction processes, developing practical biocatalytic applications of oxidoreductases has long been an imperative target in biotechnology.

#### **Acknowledgements**

The author gratefully acknowledges the Department of Chemistry, Goalpara College (Assam), India.

**23**

**Author details**

Sandhya Rani Gogoi

Department of Chemistry, Goalpara College, Goalpara, Assam, India

© 2020 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium,

\*Address all correspondence to: gogoisandhyarani@gmail.com

provided the original work is properly cited.

*Applications of Oxidoreductases*

*DOI: http://dx.doi.org/10.5772/intechopen.94409*

#### **Conflict of interest**

The author declares no conflict of interest.

*Applications of Oxidoreductases DOI: http://dx.doi.org/10.5772/intechopen.94409*

*Oxidoreductase*

cosubstrate [69]. The ClO¯

**6. Conclusions**

described that laccase-iodide salt binary iodine-generating system (for sterilization) can have several advantages over the direct iodine application [69]. Peroxidases may replace laccase for the application, even though they would require H2O2 as

Tremendous progress has been made in the recent years in the field of applica-

The author gratefully acknowledges the Department of Chemistry, Goalpara

tions of oxidoreductases. Oxidoreductases metabolism is a fundamental bioprocess that plays a pivotal role in all species, including humans, plants, animals, and microorganisms, as their specific function is to catalyze oxidation and reduction reactions that occur within the cell. Abnormality in this metabolic system leads to a number of metabolic disorders. Thus, owing to the remarkable properties of oxidoreductases, they can be used for the diagnosis of disorders. They can provide insight into the diseased state by diagnosis, prognosis, or by assessment of response therapy. It has been established that oxidoreductases as biosensors are becoming popular potential tools in biotechnology due to their high specificity. With oxidoreductases, the conversion of a variety of aliphatic/aromatic molecules can be achieved; inert hydrocarbons can be functionalized (by hydroxylation, sulfoxidation, epoxidation, etc.); regio-, enantio- (on racemic substrates); enantiotopo– (on prochiral sub-strates); and chemo-selective reactions can be accomplished; important synthons from inexpensive and renewable biomaterials can be constructed; and the negative environment impact can be reduced [69]. Since numerous chemical and biochemical transformations engage oxidation/reduction processes, developing practical biocatalytic applications of oxidoreductases has

long been an imperative target in biotechnology.

The author declares no conflict of interest.

**Acknowledgements**

College (Assam), India.

**Conflict of interest**

Mn-peroxidase are extremely effective oxidants and antimicrobial agents [70]. Peroxidase can also be used to cross-link collagen which is beneficial to the healing of damaged skin [71]. The physiological activities of lysyl oxidase comprise the extracellular matrix construction which can hasten wound-healing [72, 73]. A glucose oxidase, lactoperoxidase, and iodide system has been tested for dental care and the oxidase produces H2O2 to feed the peroxidase, so that it can produce iodine that can kill plaque-causing bacteria [74]. It has been reported that the haloperoxidase can be used to oxidatively modify rubber latex surfaces, making them less allergenic [75]. A secreted oxidoreductase may even be developed as a vaccine against secretor microbes such as, *Aspergillus oryzae* catalase A protein has been studied as a potential aspergillosis vaccine [69]. It has been reported that low-molecular-mass laccase purified from the mushroom *Tricholoma giganteumis* possesses significant HIV-1 reverse transcriptase inhibitory activity [76]. As nature's own catalysts, enzymes acquire very diverse specificity, reactivity, and other physicochemical, catalytic, and biological properties highly enviable for miscellaneous industrial and medical applications [69].

and Mn(III) species formed by haloperoxidase and

**22**

#### **Author details**

Sandhya Rani Gogoi Department of Chemistry, Goalpara College, Goalpara, Assam, India

\*Address all correspondence to: gogoisandhyarani@gmail.com

© 2020 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

#### **References**

[1] Martinez AT, Ruiz-Dueñas FJ, Camarero S, Serrano A, Linde D, Lund H, Vind J, Tovborg M, Herold-Majumdar OM, Hofrichter Mand Liers, C. Mint: Oxidoreductases on their way to industrial biotransformations. Biotechnology advances. 2017;35(6);815-831. DOI: https://doi.org/10.1016/j. biotechadv.2017.06.003

[2] Younus H. Oxidoreductases: Overview and Practical Applications. Biocatalysis: Springer, Cham; 2019. 39 p. DOI: https://doi. org/10.1007/978-3-030-25023-2\_3

[3] Singh RS, Singh T, Singh AK. Enzymes as Diagnostic Tools. Advances in Enzyme Technology: Elsevier; 2019. 225p. DOI: https://doi.org/10.1016/ B978-0-444-64114-4.00009-1

[4] Wang J, Yuan W, Chen Z, Wu S, Chen J, Ge J, Hou F, Chen Z. Mint: Overexpression of G6PD is associated with poor clinical outcome in gastric cancer. Tumor Biology. 2012;33; 95-101. DOI: https://doi.org/10.1007/ s13277-011-0251-9

[5] Chan FKM, Moriwaki K, De-Rosa MJ. Detection of necrosis by release of lactate dehydrogenase activity. In: Snow A, Lenardo M. (Eds.), Immune Homeostasis Methods and Protocols. Springer Science +Bushiness Media, New York, 2013, vol. 979. p. 65-70. DOI: https://doi. org/10.1007/978-1-62703-290-2\_7

[6] Szabo Z, Hamalainen J, Loikkanen I, Moilanen AM, Hirvikoski P, Vaisanen T, Paavonen TK, Vaarala MH. Mint: Sorbitol dehydrogenase expression is regulated by androgens in the human prostate. Oncology Reports. 2010;23;1233-1239. DOI: https://doi. org/10.3892/or\_00000755

[7] Uzozie A, Nanni P, Staiano T, Grossmann J, Barkow-Oesterreicher S, Shay JW, Tiwari A, Buffoli F, Laczko E, Marra G. Mint: Sorbitol dehydrogenase over expression and other aspects of dysregulate dprotein expression in human precancerous colorectal neoplasms: a quantitative proteomics study. Molecular & Cellular Proteomics. 2014;13;1198-1218. DOI: https://doi. org/10.1074/mcp.M113.035105

[8] Radenkovic S, Milosevic Z, Konjevic G, Karadzic K, Rovcanin B, Buta M, Gopcevic K, Jurisic V. Mint: Lactate dehydrogenase, catalase and superoxide dismutase in tumor tissue of breast cancer patients in respect to mammographic findings. Cell Biochemistry and Biophysics. 2013;66;287-295. DOI: https://doi. org/10.1007/s12013-012-9482-7

[9] Koukourakis MI, Giatromanolaki A, Sivridis E, Gatter KC, Trarbach T, Folprecht G, Shi MM, Lebwohl D, Jalava T, Laurent D, Meinhardt G. Mint: Prognostic and predictive role of lactate dehydrogenase 5 expression in colorectal cancer patients treated with PTK787/ZK 222584 (vatalanib) antiangiogenic therapy. Clinical Cancer Research. 2011;17;4892-4900. DOI: 10.1158/1078- 0432.CCR-10-2918

[10] Hermes A, Gatzemeier U, Waschki B, Reck M. Mint: Lactate dehydrogenase as prognostic factor in limited and extensive disease stage small cell lung cancer - a retrospective single institution analysis. Respiratory Medicine. 2010;104;1937-1942. DOI: https://doi.org/10.1016/j. rmed.2010.07.013

[11] Hsieh AH, Tahkar H, Koczwara B, Kichenadasse G, Beckmann K, Karapetis C, Sukumaran S. Mint: Pretreatment serum lactate dehydrogenase as a biomarker in small cell lung cancer. Asia-Pacific Journal of Clinical Oncology. 2018;14(2);e64-70. DOI: https://doi.org/10.1111/ajco.12674

**25**

*Applications of Oxidoreductases*

net/publication/321028729

[13] Brown JE, Cook RJ, Lipton A, Coleman RE. Mint: Serum lactate dehydrogenase is prognostic for survival in patients with bone metastases from breast cancer: a retrospective analysis in bisphosphonate-treated patients. Clinical Cancer Research. 2012;18;6348-

6355. DOI: 10.1158/1078-0432.

[14] Nandita A, Basavaraju SM, Pachipulusu B. Mint: Lactate

dehydrogenase as a tumor marker in oral cancer and oral potentially malignant disorders: a biochemical study International Journal of Preventive & Clinical Dental Research. 2017;4;1-5. DOI: 10.5005/jp-journals-10052-0108

[15] Halabi S, Small EJ, Kantoff PW, Kattan MW, Kaplan EB, Dawson NA,

Vogelzang NJ. Mint: Prognostic model for predicting survival in men with hormone-refractory metastatic prostate cancer. Journal of Clinical Oncology. 2003;21;1232-1237. DOI: 10.1200/

[16] Gerlinger M, Wilson P, Powles T, Shamash J. Mint: Elevated LDH predicts

poor outcome of recurrent germ cell tumours treated with dose dense chemotherapy. European Journal of Cancer. 2010;46;2913- 2918. DOI: https://doi.org/10.1016/j.

[17] Rong Y, Wu W, Ni X, Kuang T, Jin D, Wang D, Lou W. Mint: Lactate dehydrogenase A is over expressed in pancreatic cancer and promotes the growth of pancreatic cancer cells. Tumor Biology. 2013;34;1523-1530.

Levine EG, Blumenstein BA,

JCO.2003.06.100

ejca.2010.07.004

CCR-12-1397

*DOI: http://dx.doi.org/10.5772/intechopen.94409*

[12] Zheng X, Wang K, Xu L, Ye P, Cai S, Lu H, Bao C, Kong J. Mint: The effect of serum lactate dehydrogenase levels on lung cancer prognosis: ameta-analysis. International Journal of Clinical and Experimental Medicine. 2017;10;14179- 14186. DOI: https://www.researchgate.

DOI: https://doi.org/10.1007/

[18] Walaa-Fikry ME. Mint: Lactate dehydrogenase (LDH) as prognostic marker in acute leukemia "Quantitative Method". Journal of Blood Disorders Transfusion. 2017;8;1-9. DOI: 10.4172/2155-9864.1000375

[19] Kato GJ, Nouraie SM, Gladwin MT. Mint: Lactate dehydrogenase and hemolysis in sickle cell disease, Blood. 2013;122;1091-1092. DOI: https://doi. org/10.1182/blood-2013-05-505016

Wilson GS. Biosensors: Fundamentals and Applications. 1st ed. Oxford University Press, Oxford, 1987. DOI: https://www.diva-portal.org/smash/get/

[21] Rocchitta G, Spanu A, Babudieri S, Latte G, Madeddu G, Galleri G, Nuvoli S, Bagella P, Demartis MI, Fiore V, Manetti R. Mint: Enzyme biosensors for biomedical applications: Strategies for safeguarding analytical performances in biological fluids. Sensors. 2016;16(6);780-801. DOI: https://doi.org/10.3390/s16060780

diva2:619968/FULLTEXT01.pdf

[22] Schmidt HL, Schuhmann W. Mint: Reagentless oxidoreductase sensors. Biosensors and Bioelectronics, 1996;11(1-2);127-135. DOI: https://doi. org/10.1016/0956-5663(96)83720-1

[23] Laurinavicius V, Kurtinaitiene B, Liauksminas V, Ramanavicius A, Meskys R, Rudomanskis R, Skotheim T, Boguslavsky L. Mint: Oxygen insensitive

glucose biosensor based on PQQ-

[24] Bollella P, Gorton L, Ludwig R, Antiochia R. Mint: A third generation glucose biosensor based on cellobiosedehydrogenase immobilized on a glassy carbon electrode decorated with electrodeposited gold nanoparticles:

dependent glucose dehydrogenase. Anal. Lett. 1999;32;299-316. DOI: https://doi. org/10.1080/00032719908542822

[20] Turner APF, Karube I,

s13277-013-0679-1

#### *Applications of Oxidoreductases DOI: http://dx.doi.org/10.5772/intechopen.94409*

[12] Zheng X, Wang K, Xu L, Ye P, Cai S, Lu H, Bao C, Kong J. Mint: The effect of serum lactate dehydrogenase levels on lung cancer prognosis: ameta-analysis. International Journal of Clinical and Experimental Medicine. 2017;10;14179- 14186. DOI: https://www.researchgate. net/publication/321028729

[13] Brown JE, Cook RJ, Lipton A, Coleman RE. Mint: Serum lactate dehydrogenase is prognostic for survival in patients with bone metastases from breast cancer: a retrospective analysis in bisphosphonate-treated patients. Clinical Cancer Research. 2012;18;6348- 6355. DOI: 10.1158/1078-0432. CCR-12-1397

[14] Nandita A, Basavaraju SM, Pachipulusu B. Mint: Lactate dehydrogenase as a tumor marker in oral cancer and oral potentially malignant disorders: a biochemical study International Journal of Preventive & Clinical Dental Research. 2017;4;1-5. DOI: 10.5005/jp-journals-10052-0108

[15] Halabi S, Small EJ, Kantoff PW, Kattan MW, Kaplan EB, Dawson NA, Levine EG, Blumenstein BA, Vogelzang NJ. Mint: Prognostic model for predicting survival in men with hormone-refractory metastatic prostate cancer. Journal of Clinical Oncology. 2003;21;1232-1237. DOI: 10.1200/ JCO.2003.06.100

[16] Gerlinger M, Wilson P, Powles T, Shamash J. Mint: Elevated LDH predicts poor outcome of recurrent germ cell tumours treated with dose dense chemotherapy. European Journal of Cancer. 2010;46;2913- 2918. DOI: https://doi.org/10.1016/j. ejca.2010.07.004

[17] Rong Y, Wu W, Ni X, Kuang T, Jin D, Wang D, Lou W. Mint: Lactate dehydrogenase A is over expressed in pancreatic cancer and promotes the growth of pancreatic cancer cells. Tumor Biology. 2013;34;1523-1530.

DOI: https://doi.org/10.1007/ s13277-013-0679-1

[18] Walaa-Fikry ME. Mint: Lactate dehydrogenase (LDH) as prognostic marker in acute leukemia "Quantitative Method". Journal of Blood Disorders Transfusion. 2017;8;1-9. DOI: 10.4172/2155-9864.1000375

[19] Kato GJ, Nouraie SM, Gladwin MT. Mint: Lactate dehydrogenase and hemolysis in sickle cell disease, Blood. 2013;122;1091-1092. DOI: https://doi. org/10.1182/blood-2013-05-505016

[20] Turner APF, Karube I, Wilson GS. Biosensors: Fundamentals and Applications. 1st ed. Oxford University Press, Oxford, 1987. DOI: https://www.diva-portal.org/smash/get/ diva2:619968/FULLTEXT01.pdf

[21] Rocchitta G, Spanu A, Babudieri S, Latte G, Madeddu G, Galleri G, Nuvoli S, Bagella P, Demartis MI, Fiore V, Manetti R. Mint: Enzyme biosensors for biomedical applications: Strategies for safeguarding analytical performances in biological fluids. Sensors. 2016;16(6);780-801. DOI: https://doi.org/10.3390/s16060780

[22] Schmidt HL, Schuhmann W. Mint: Reagentless oxidoreductase sensors. Biosensors and Bioelectronics, 1996;11(1-2);127-135. DOI: https://doi. org/10.1016/0956-5663(96)83720-1

[23] Laurinavicius V, Kurtinaitiene B, Liauksminas V, Ramanavicius A, Meskys R, Rudomanskis R, Skotheim T, Boguslavsky L. Mint: Oxygen insensitive glucose biosensor based on PQQdependent glucose dehydrogenase. Anal. Lett. 1999;32;299-316. DOI: https://doi. org/10.1080/00032719908542822

[24] Bollella P, Gorton L, Ludwig R, Antiochia R. Mint: A third generation glucose biosensor based on cellobiosedehydrogenase immobilized on a glassy carbon electrode decorated with electrodeposited gold nanoparticles:

**24**

*Oxidoreductase*

**References**

[1] Martinez AT, Ruiz-Dueñas FJ, Camarero S, Serrano A, Linde D, Lund H, Vind J, Tovborg M, Herold-Majumdar OM, Hofrichter Mand Liers, C. Mint: Oxidoreductases Shay JW, Tiwari A, Buffoli F, Laczko E, Marra G. Mint: Sorbitol dehydrogenase over expression and other aspects of dysregulate dprotein expression in human precancerous colorectal neoplasms: a quantitative proteomics study. Molecular & Cellular Proteomics. 2014;13;1198-1218. DOI: https://doi. org/10.1074/mcp.M113.035105

[8] Radenkovic S, Milosevic Z, Konjevic G, Karadzic K, Rovcanin B, Buta M, Gopcevic K, Jurisic V. Mint: Lactate dehydrogenase, catalase and superoxide dismutase in tumor tissue of breast cancer patients in respect to mammographic findings. Cell Biochemistry and Biophysics. 2013;66;287-295. DOI: https://doi. org/10.1007/s12013-012-9482-7

[9] Koukourakis MI,

0432.CCR-10-2918

rmed.2010.07.013

Giatromanolaki A, Sivridis E, Gatter KC, Trarbach T, Folprecht G, Shi MM, Lebwohl D, Jalava T, Laurent D, Meinhardt G. Mint: Prognostic and predictive role of lactate dehydrogenase 5 expression in colorectal cancer patients treated with PTK787/ZK 222584 (vatalanib) antiangiogenic therapy. Clinical Cancer Research. 2011;17;4892-4900. DOI: 10.1158/1078-

[10] Hermes A, Gatzemeier U, Waschki B, Reck M. Mint: Lactate dehydrogenase as prognostic factor in limited and extensive disease stage small cell lung cancer - a retrospective single institution analysis. Respiratory Medicine. 2010;104;1937-1942. DOI: https://doi.org/10.1016/j.

[11] Hsieh AH, Tahkar H, Koczwara B,

Kichenadasse G, Beckmann K, Karapetis C, Sukumaran S. Mint: Pretreatment serum lactate dehydrogenase as a biomarker in small cell lung cancer. Asia-Pacific Journal of Clinical Oncology. 2018;14(2);e64-70. DOI: https://doi.org/10.1111/ajco.12674

biotransformations. Biotechnology advances. 2017;35(6);815-831. DOI: https://doi.org/10.1016/j. biotechadv.2017.06.003

[2] Younus H. Oxidoreductases: Overview and Practical Applications.

Biocatalysis: Springer, Cham; 2019. 39 p. DOI: https://doi. org/10.1007/978-3-030-25023-2\_3

[3] Singh RS, Singh T, Singh AK.

[4] Wang J, Yuan W, Chen Z,

[5] Chan FKM, Moriwaki K, De-Rosa MJ. Detection of necrosis by release of lactate dehydrogenase activity. In: Snow A, Lenardo M. (Eds.), Immune Homeostasis Methods and Protocols. Springer Science +Bushiness Media, New York, 2013, vol. 979. p. 65-70. DOI: https://doi. org/10.1007/978-1-62703-290-2\_7

[6] Szabo Z, Hamalainen J,

org/10.3892/or\_00000755

is regulated by androgens in the human prostate. Oncology Reports. 2010;23;1233-1239. DOI: https://doi.

[7] Uzozie A, Nanni P, Staiano T, Grossmann J, Barkow-Oesterreicher S,

Loikkanen I, Moilanen AM, Hirvikoski P, Vaisanen T, Paavonen TK, Vaarala MH. Mint: Sorbitol dehydrogenase expression

s13277-011-0251-9

Enzymes as Diagnostic Tools. Advances in Enzyme Technology: Elsevier; 2019. 225p. DOI: https://doi.org/10.1016/ B978-0-444-64114-4.00009-1

Wu S, Chen J, Ge J, Hou F, Chen Z. Mint: Overexpression of G6PD is associated with poor clinical outcome in gastric cancer. Tumor Biology. 2012;33; 95-101. DOI: https://doi.org/10.1007/

on their way to industrial

characterization and application in human saliva. Sensors. 2017;17;1912- 1926. DOI: https://doi.org/10.3390/ s17081912

[25] Clemens AH, Chang PH, Myers RW. Mint: Development of an automatic system of insulin infusion controlled by blood sugar, its system for the determination of glucose and control algorithms. Journees annuelles de diabetologie de l'Hotel-Dieu. 1976; 269- 278. DOI: https://pubmed.ncbi.nlm.nih. gov/1011418/

[26] Yao H, Li N, Xu JZ, Zhu JJ. Mint: A glucose biosensor based on immobilization of glucose oxidase in chitosan network matrix. Chinese Journal Chemistry. 2005;23;275- 279. DOI: https://doi.org/10.1002/ cjoc.200590275

[27] Chu X, Wu B, Xiao C, Zhang X, Chen J. Mint: A new amperometric glucose biosensor based on platinum nanoparticles/polymerized ionic liquidcarbon nanotubes nanocomposites. Electrochimica Acta. 2010;55;2848- 2852. DOI: https://doi.org/10.1016/j. electacta.2009.12.057

[28] Periasamy AP, Chang YJ, Chen SM. Mint: Amperometric glucose sensor based on glucose oxidase immobilized on gelatin-multiwalled carbon nanotube modified glassy carbon electrode. Bioelectrochemistry. 2011;80;114- 120. DOI: https://doi.org/10.1016/j. bioelechem.2010.06.009

[29] Qiu C, Wang X, Liu X, Hou S, Ma H. Mint: Direct electrochemistry of glucoseoxidase immobilized on nano structured gold thin films and its application to bioelectrochemical glucose sensor. Electrochimica Acta. 2012;67;140-146. DOI: https://doi. org/10.1016/j.electacta.2012.02.011

[30] Yadav S, Devi R, Kumari S, Yadav S, Pundir CS. Mint: An amperometric oxalate biosensor based on sorghum

oxalate oxidase bound carboxylated multiwalled carbon nanotubespolyaniline composite film. Journal of Biotechnology. 2011;151;212-217. DOI: https://doi.org/10.1016/j. jbiotec.2010.12.008

[31] Nandini S, Nalini S, Reddy MM, Suresh GS, Melo JS, Niranjana P, Sanetuntikul J, Shanmugam S. Mint: Synthesis of one-dimensional gold nanostructures and the electrochemical application of the nanohybrid containing functionalized graphene oxide for cholesterol biosensing. Bioelectrochemistry. 2016;110;79- 90. DOI: https://doi.org/10.1016/j. bioelechem.2016.03.006

[32] Pakapongpan S, Tuantranont A, Sritongkham P. Mint: Cholesterol biosensor based on direct electron transfer of cholesterol oxidase on multi-wall carbon nanotubes. IEEE. 2011;2011;138-141. DOI: 10.1109/ BMEiCon.2012.6172037

[33] Pundir CS, Narang J, Chauhan N, Sharma P, Sharma R. Mint: An amperometric cholesterol biosensor based on epoxy resin membrane bound cholesterol oxidase. Indian Journal of Medical Research. 2012;136;633-640. DOI: https://pubmed.ncbi.nlm.nih. gov/23168704/

[34] Sekretaryova AN, Beni V, Eriksson M, Karyakin AA, Turner AP, Vagin MY. Mint: Cholesterol selfpowered biosensor. Analytical Chemistry. 2014;86;9540-9547. DOI: https://doi.org/10.1021/ac501699p

[35] Lupu A, Valsesia A, Bretagnol F, Colpo P, Rossi F. Mint: Development of a potentiometric biosensor based on nanostructured surface for lactate determination. Sensors Actuators B: Chemical. 2007;127;606-612. DOI: https://doi.org/10.1016/j. snb.2007.05.020

[36] Ibupoto ZH, Shah SMUA, Khun K, Willander M. Mint: Electrochemical

**27**

*Applications of Oxidoreductases*

L-lactic acid sensor based on immobilized ZnO nanorods with lactate oxidase. Sensors. 2012;12;2456- 2466. DOI: https://doi.org/10.3390/

s120302456

*DOI: http://dx.doi.org/10.5772/intechopen.94409*

Mint: The BRENDA enzyme

jbiotec.2017.04.020

b98911

C3GC37129H

information system–From a database to an expert system. Journal of Biotechnology. 2017; 261;194-206. DOI: https://doi.org/10.1016/j.

[43] Wichmann R, Vasic-Racki D.

[44] Wu H, Tian C, Song X, Liu C, Yang D, Jiang Z. Mint: Methods for the regeneration of nicotinamide coenzymes. Green Chemistry. 2013;15(7);1773-1789. DOI: 10.1039/

[45] Weckbecker A, Gröger H, Hummel W. Regeneration of nicotinamide coenzymes: principles and applications for the synthesis of chiral compounds. In Biosystems Engineering I. Springer, Berlin, Heidelberg; 2010. p. 195-242. DOI: https://doi.org/10.1007/10\_2009\_55

[46] Tishkov VI, Popov VO. Mint: Protein engineering of formate dehydrogenase. Biomolecular Engineering. 2006;23;89-110. DOI: https://doi.org/10.1016/j.

[47] Hoelsch K, Sührer I, Heusel M, Weuster-Botz D. Mint: Engineering of formate dehydrogenase: Synergistic effect of mutations affecting cofactor specificity and chemical stability. Applied Microbiology and Biotechnology. 2013;97;2473-2481. DOI:

10.1007/s00253-012-4142-9

org/10.1002/bit.21168

[48] Johannes TW, Woodyer RD, Zhao H. Mint: Efficient regeneration of NADPH using an engineered phosphite dehydrogenase.

Biotechnology and Bioengineering. 2007;96;18-26. DOI: https://doi.

bioeng.2006.02.003

Cofactor regeneration at the lab scale. In: Technology transfer in biotechnology. Springer, Berlin, Heidelberg; 2005. p. 225-260. DOI: https://doi.org/10.1007/

[37] Haghighi B, Bozorgzadeh S. Mint: Fabrication of a highly sensitive electro chemiluminescence lactate biosensor using ZnO nanoparticles decorated multiwalled carbon nanotubes. Talanta. 2011;85;2189-2193. DOI: https://doi. org/10.1016/j.talanta.2011.07.071

[38] Jiang D, Chu Z, Peng J, Jin W. Mint: Screen-printed biosensor chips with Prussian blue nanocubes for the detection of physiological analytes. Sensors Actuators B: Chemical. 2016;228;679-687. DOI: https://doi. org/10.1016/j.snb.2016.01.076

[39] Romero MR, Garay F, Baruzzi AM. Mint: Design and optimization of a lactate amperometric biosensor based on lactate oxidase cross-linked with polymeric matrixes. Sensors Actuators B: Chemical. 2008;131;590- 595. DOI: https://doi.org/10.1016/j.

snb.2007.12.044

bios.2015.01.044

[40] Briones M, Casero E, Petit-Dominguez MD, Ruiz MA, Parra-Alfambra AM, Pariente F, Lorenzo E, Vazquez L. Mint: Diamond

nanoparticles based biosensors for efficient glucose and lactate determination. Biosensors and Bioelectronics. 2015;68;521-528. DOI: https://doi.org/10.1016/j.

[41] You C, Huang R, Wei X, Zhu Z, Zhang YHP. Mint: Protein engineering

nicotinamide-based coenzymes, with applications in synthetic biology. Synthetic and Systems Biotechnology, 2017;2(3);208-218. DOI: https://doi. org/10.1016/j.synbio.2017.09.002

[42] Schomburg I, Jeske L, Ulbrich M, Placzek S, Chang A, Schomburg D.

of oxidoreductases utilizing

#### *Applications of Oxidoreductases DOI: http://dx.doi.org/10.5772/intechopen.94409*

*Oxidoreductase*

s17081912

gov/1011418/

cjoc.200590275

electacta.2009.12.057

bioelechem.2010.06.009

[29] Qiu C, Wang X, Liu X, Hou S, Ma H. Mint: Direct electrochemistry of glucoseoxidase immobilized on nano structured gold thin films and its application to bioelectrochemical glucose sensor. Electrochimica Acta. 2012;67;140-146. DOI: https://doi. org/10.1016/j.electacta.2012.02.011

[30] Yadav S, Devi R, Kumari S, Yadav S, Pundir CS. Mint: An amperometric oxalate biosensor based on sorghum

[26] Yao H, Li N, Xu JZ, Zhu JJ. Mint: A glucose biosensor based on immobilization of glucose oxidase in chitosan network matrix. Chinese Journal Chemistry. 2005;23;275- 279. DOI: https://doi.org/10.1002/

[27] Chu X, Wu B, Xiao C, Zhang X, Chen J. Mint: A new amperometric glucose biosensor based on platinum nanoparticles/polymerized ionic liquidcarbon nanotubes nanocomposites. Electrochimica Acta. 2010;55;2848- 2852. DOI: https://doi.org/10.1016/j.

[28] Periasamy AP, Chang YJ, Chen SM. Mint: Amperometric glucose sensor based on glucose oxidase immobilized on gelatin-multiwalled carbon nanotube modified glassy carbon electrode. Bioelectrochemistry. 2011;80;114- 120. DOI: https://doi.org/10.1016/j.

characterization and application in human saliva. Sensors. 2017;17;1912- 1926. DOI: https://doi.org/10.3390/

oxalate oxidase bound carboxylated multiwalled carbon nanotubespolyaniline composite film. Journal of Biotechnology. 2011;151;212-217. DOI: https://doi.org/10.1016/j.

[31] Nandini S, Nalini S, Reddy MM, Suresh GS, Melo JS, Niranjana P, Sanetuntikul J, Shanmugam S. Mint: Synthesis of one-dimensional gold nanostructures and the electrochemical

[32] Pakapongpan S, Tuantranont A, Sritongkham P. Mint: Cholesterol biosensor based on direct electron transfer of cholesterol oxidase on multi-wall carbon nanotubes. IEEE. 2011;2011;138-141. DOI: 10.1109/

[33] Pundir CS, Narang J, Chauhan N, Sharma P, Sharma R. Mint: An amperometric cholesterol biosensor based on epoxy resin membrane bound cholesterol oxidase. Indian Journal of Medical Research. 2012;136;633-640. DOI: https://pubmed.ncbi.nlm.nih.

application of the nanohybrid containing functionalized graphene oxide for cholesterol biosensing. Bioelectrochemistry. 2016;110;79- 90. DOI: https://doi.org/10.1016/j.

bioelechem.2016.03.006

BMEiCon.2012.6172037

gov/23168704/

snb.2007.05.020

[34] Sekretaryova AN, Beni V,

Eriksson M, Karyakin AA, Turner AP, Vagin MY. Mint: Cholesterol selfpowered biosensor. Analytical Chemistry. 2014;86;9540-9547. DOI: https://doi.org/10.1021/ac501699p

[35] Lupu A, Valsesia A, Bretagnol F, Colpo P, Rossi F. Mint: Development of a potentiometric biosensor based on nanostructured surface for lactate determination. Sensors Actuators B: Chemical. 2007;127;606-612. DOI: https://doi.org/10.1016/j.

[36] Ibupoto ZH, Shah SMUA, Khun K, Willander M. Mint: Electrochemical

jbiotec.2010.12.008

[25] Clemens AH, Chang PH, Myers RW. Mint: Development of an automatic system of insulin infusion controlled by blood sugar, its system for the determination of glucose and control algorithms. Journees annuelles de diabetologie de l'Hotel-Dieu. 1976; 269- 278. DOI: https://pubmed.ncbi.nlm.nih.

**26**

L-lactic acid sensor based on immobilized ZnO nanorods with lactate oxidase. Sensors. 2012;12;2456- 2466. DOI: https://doi.org/10.3390/ s120302456

[37] Haghighi B, Bozorgzadeh S. Mint: Fabrication of a highly sensitive electro chemiluminescence lactate biosensor using ZnO nanoparticles decorated multiwalled carbon nanotubes. Talanta. 2011;85;2189-2193. DOI: https://doi. org/10.1016/j.talanta.2011.07.071

[38] Jiang D, Chu Z, Peng J, Jin W. Mint: Screen-printed biosensor chips with Prussian blue nanocubes for the detection of physiological analytes. Sensors Actuators B: Chemical. 2016;228;679-687. DOI: https://doi. org/10.1016/j.snb.2016.01.076

[39] Romero MR, Garay F, Baruzzi AM. Mint: Design and optimization of a lactate amperometric biosensor based on lactate oxidase cross-linked with polymeric matrixes. Sensors Actuators B: Chemical. 2008;131;590- 595. DOI: https://doi.org/10.1016/j. snb.2007.12.044

[40] Briones M, Casero E, Petit-Dominguez MD, Ruiz MA, Parra-Alfambra AM, Pariente F, Lorenzo E, Vazquez L. Mint: Diamond nanoparticles based biosensors for efficient glucose and lactate determination. Biosensors and Bioelectronics. 2015;68;521-528. DOI: https://doi.org/10.1016/j. bios.2015.01.044

[41] You C, Huang R, Wei X, Zhu Z, Zhang YHP. Mint: Protein engineering of oxidoreductases utilizing nicotinamide-based coenzymes, with applications in synthetic biology. Synthetic and Systems Biotechnology, 2017;2(3);208-218. DOI: https://doi. org/10.1016/j.synbio.2017.09.002

[42] Schomburg I, Jeske L, Ulbrich M, Placzek S, Chang A, Schomburg D.

Mint: The BRENDA enzyme information system–From a database to an expert system. Journal of Biotechnology. 2017; 261;194-206. DOI: https://doi.org/10.1016/j. jbiotec.2017.04.020

[43] Wichmann R, Vasic-Racki D. Cofactor regeneration at the lab scale. In: Technology transfer in biotechnology. Springer, Berlin, Heidelberg; 2005. p. 225-260. DOI: https://doi.org/10.1007/ b98911

[44] Wu H, Tian C, Song X, Liu C, Yang D, Jiang Z. Mint: Methods for the regeneration of nicotinamide coenzymes. Green Chemistry. 2013;15(7);1773-1789. DOI: 10.1039/ C3GC37129H

[45] Weckbecker A, Gröger H, Hummel W. Regeneration of nicotinamide coenzymes: principles and applications for the synthesis of chiral compounds. In Biosystems Engineering I. Springer, Berlin, Heidelberg; 2010. p. 195-242. DOI: https://doi.org/10.1007/10\_2009\_55

[46] Tishkov VI, Popov VO. Mint: Protein engineering of formate dehydrogenase. Biomolecular Engineering. 2006;23;89-110. DOI: https://doi.org/10.1016/j. bioeng.2006.02.003

[47] Hoelsch K, Sührer I, Heusel M, Weuster-Botz D. Mint: Engineering of formate dehydrogenase: Synergistic effect of mutations affecting cofactor specificity and chemical stability. Applied Microbiology and Biotechnology. 2013;97;2473-2481. DOI: 10.1007/s00253-012-4142-9

[48] Johannes TW, Woodyer RD, Zhao H. Mint: Efficient regeneration of NADPH using an engineered phosphite dehydrogenase. Biotechnology and Bioengineering. 2007;96;18-26. DOI: https://doi. org/10.1002/bit.21168

[49] Woodyer R, Van der Donk WA, Zhao H. Mint: Relaxing the nicotinamide cofactor specificity of phosphite dehydrogenase by rational design. Biochemistry. 2003;42;11604- 11614. DOI: https://doi.org/10.1021/ bi035018b

[50] Wong C-H, Drueckhammer DG, Sweers HM. Mint: Enzymatic vs. fermentative synthesis: Thermostable glucose dehydrogenase catalyzed regeneration of NAD(P)H for use in enzymatic synthesis. Journal of the American Chemical Society. 1985;107;4028-4031. DOI: https://doi. org/10.1021/ja00299a044

[51] Kaswurm V, Hecke WV, Kulbe KD, Ludwig R. Mint: Guidelines for the application of NAD(P)H regenerating glucose dehydrogenase in synthetic processes. Advanced Synthesis and Catalysis. 2013;355;1709-1714. DOI: https://doi.org/10.1002/adsc.201200959

[52] Johannes TW, Woodyer RD, Zhao H. Mint: Efficient regeneration of NADPH using an engineered phosphite dehydrogenase. Biotechnology and Bioengineering. 2007;96(1);18-26. DOI: https://doi.org/10.1002/bit.21168

[53] Petschacher B, Staunig N, Müller M, Schürmann M, Mink D, De Wildeman S, Gruber K, Glieder A. Mint: Cofactor specificity engineering of Streptococcus mutans NADH oxidase 2 for NAD(P)+ regeneration in biocatalytic oxidations. Computational and Structural Biotechnology Journal. 2014;9(14);e201402005. DOI: https:// doi.org/10.5936/csbj.201402005

[54] Hollmann F, Arends IW, Buehler K, Schallmey A, Bühler B. Mint: Enzymemediated oxidations for the chemist. Green Chemistry. 2011;13(2);226-265. DOI: 10.1039/C0GC00595A

[55] Monti D, Ottolina G, Carrea G, Riva S. Mint: Redox reactions catalyzed by isolated enzymes. Chemical reviews, 2011;111(7);4111-4140. DOI: 10.1021/ cr100334x

[56] Romano D, Villa R, Molinari F. Mint: Preparative biotransformations: oxidation of alcohols. ChemCatChem. 2012;4(6);739-749. DOI: https://doi. org/10.1002/cctc.201200042

[57] Uyama H. Synthesis of Poly(aromatic)s I: Oxidoreductase as Catalyst. In Enzymatic Polymerization towards Green Polymer Chemistry. Springer, Singapore. 2019. p. 267-305. DOI: https://doi. org/10.1007/978-981-13-3813-7\_9

[58] Kobayashi S. Mint: Enzymatic polymerization: a new method of polymer synthesis. Journal of Polymer Science Part A: Polymer Chemistry. 1999; 37(16);3041-3056. DOI: https://doi.org/10.1002/(SICI)1099- 0518(19990815)37:16<3041::AID-POLA1>3.0.CO;2-V

[59] Ćirić-Marjanović G, Milojević-Rakić M, Janošević-Ležaić A, Luginbühl S, Walde P. Mint: Enzymatic oligomerization and polymerization of arylamines: state of the art and perspectives. Chemical Papers. 2017;71(2);199-242. DOI: 10.1007/ s11696-016-0094-3

[60] de Gonzalo G, A Orden, A, R Bisogno F. Mint: New trends in organic synthesis with oxidative enzymes. Current Organic Chemistry. 2012;16(21);2598-2612. DOI: https://doi. org/10.2174/138527212804004599

[61] Kroutil W, Mang H, Edegger K, Faber K. Mint: Biocatalytic oxidation of primary and secondary alcohols. Advanced Synthesis & Catalysis. 2004;346(2-3);125-142. DOI: https:// doi.org/10.1002/adsc.200303177

[62] Turner NJ. Mint: Enantioselective oxidation of C–O and C–N bonds using oxidases. Chemical Reviews.

**29**

*Applications of Oxidoreductases*

org/10.1021/cr200111v

*DOI: http://dx.doi.org/10.5772/intechopen.94409*

2011;111(7);4073-4087. DOI: https://doi.

[69] Xu F. Mint: Applications of oxidoreductases: recent progress. Industrial Biotechnology. 2005; 1;1(1);38-50. DOI: https://doi. org/10.1089/ind.2005.1.38

[71] Miller DR, Tizard IR,

WO200135882-A(2001).

US6808707-B2(2004).

[70] Danielsen S, Christensen BE. PCT patent WO2003047351-A(2003).

Keeton JT, Prochaska JF. PCT patent

[73] Perrier E, Cenizo V, Bouez C, Sommer P, Damour O, Gleyzal C, Andre V, Reymermier C, inventors; Centre National de la Recherche Scientifique CNRS, Coletica, assignee.

Stimulation of the synthesis of the activity of an isoform of lysyl oxidase-like LOXL for stimulating the formation of elastic fibres. United States patent application US 10/852,065. 2004 Dec 16. US patent

US2004253220-A1(2004).

[74] Szynol A, De Soet JJ, Siebenvan Tuyl E, Bos JW, Frenken LG. Mint: Bactericidal effects of a fusion

antibodies coupled to glucose oxidase on oral bacteria. Antimicrobial Agents and Chemotherapy. 2004;;48(9):3390-3395.

[75] Novozymes AS. Danish patent

Purification of a novel low-molecularmass laccase with HIV-1 reverse transcriptase inhibitory activity from the mushroom Tricholoma giganteum. Biochemical and Biophysical Research Communications. 2004;315(2);450-4. DOI: 10.1016/j.bbrc.2004.01.064

DK200100630-A (2001).

[76] Wang HX, Ng TB. Mint:

protein of llama heavy-chain

[72] Ensley BD, inventor; Matrix Design, assignee. Wound healing compositions and methods using tropoelastin and lysyl oxidase. United States patent US 6,808,707. 2004 Oct 26. US patent

Schmidt S, Wang Y, Younes S, Zhang W. Mint: Biocatalytic oxidation reactions: A chemist's perspective. Angewandte Chemie International Edition. 2018; 57(30);9238-9261. DOI: https://doi. org/10.1002/anie.201800343

[64] Mifsud M, Gargiulo S, Iborra S, Arends IW, Hollmann F, Corma A. Mint: Photobiocatalytic chemistry of oxidoreductases using water as the electron donor. Nature

Communications. 2014; 5(1);1-6. DOI:

[65] Paul PEV, Sangeetha V, Deepika RG. Emerging trends in the industrial production of chemical products by microorganisms. In: Recent

developments in applied microbiology and biochemistry. Academic Press. 2019. p. 107-125. DOI: https://doi.org/10.1016/

Müller M. Mint: Biocatalytic reduction

B978-0-12-816328-3.00009-X

[66] Wolberg M, Hummel W,

of β, δ-diketo esters: A highly stereoselective approach to all four stereoisomers of a chlorinated β, δ-dihydroxy hexanoate. Chemistry–A European Journal. 2001;5;7(21);4562-71. DOI: https://doi.org/10.1002/1521- 3765(20011105)7:21<4562::AID-

CHEM4562>3.0.CO;2-4

Sons,New York, 1999).

10.1038/nature01454

[67] Xu F. in The Encyclopedia of Bioprocessing Technology: Fermentation, Biocatalysis,and

[68] Ose T, Watanabe K, Mie T, Honma M, Watanabe H, Yao M, Oikawa H, Tanaka I. Mint: Insight into a natural Diels–Alder reaction from the structure of macrophomate synthase. Nature. 2003;422(6928);185-9. DOI:

Bioseparation, eds. Flickinger MC, and Drew SW. 1545-1554 (John Wiley &

10.1038/ncomms4145

[63] Dong J, Fernández-Fueyo E, Hollmann F, Paul CE, Pesic M,

*Applications of Oxidoreductases DOI: http://dx.doi.org/10.5772/intechopen.94409*

*Oxidoreductase*

bi035018b

[49] Woodyer R, Van der Donk WA,

2011;111(7);4111-4140. DOI: 10.1021/

[56] Romano D, Villa R, Molinari F. Mint: Preparative biotransformations: oxidation of alcohols. ChemCatChem. 2012;4(6);739-749. DOI: https://doi.

Poly(aromatic)s I: Oxidoreductase

Chemistry. Springer, Singapore. 2019. p. 267-305. DOI: https://doi. org/10.1007/978-981-13-3813-7\_9

[58] Kobayashi S. Mint: Enzymatic polymerization: a new method of polymer synthesis. Journal of Polymer Science Part A: Polymer Chemistry. 1999; 37(16);3041-3056. DOI: https://doi.org/10.1002/(SICI)1099- 0518(19990815)37:16<3041::AID-

[59] Ćirić-Marjanović G, Milojević-Rakić M, Janošević-Ležaić A,

[60] de Gonzalo G, A Orden, A, R Bisogno F. Mint: New trends in organic synthesis with oxidative enzymes. Current Organic Chemistry. 2012;16(21);2598-2612. DOI: https://doi. org/10.2174/138527212804004599

[61] Kroutil W, Mang H, Edegger K, Faber K. Mint: Biocatalytic oxidation of primary and secondary alcohols. Advanced Synthesis & Catalysis. 2004;346(2-3);125-142. DOI: https:// doi.org/10.1002/adsc.200303177

[62] Turner NJ. Mint: Enantioselective oxidation of C–O and C–N bonds using oxidases. Chemical Reviews.

Luginbühl S, Walde P. Mint: Enzymatic oligomerization and polymerization of arylamines: state of the art and perspectives. Chemical Papers. 2017;71(2);199-242. DOI: 10.1007/

Polymerization towards Green Polymer

org/10.1002/cctc.201200042

[57] Uyama H. Synthesis of

as Catalyst. In Enzymatic

POLA1>3.0.CO;2-V

s11696-016-0094-3

cr100334x

nicotinamide cofactor specificity of phosphite dehydrogenase by rational design. Biochemistry. 2003;42;11604- 11614. DOI: https://doi.org/10.1021/

[50] Wong C-H, Drueckhammer DG, Sweers HM. Mint: Enzymatic vs. fermentative synthesis: Thermostable glucose dehydrogenase catalyzed regeneration of NAD(P)H for use in enzymatic synthesis. Journal of the American Chemical Society. 1985;107;4028-4031. DOI: https://doi.

[51] Kaswurm V, Hecke WV, Kulbe KD, Ludwig R. Mint: Guidelines for the application of NAD(P)H regenerating glucose dehydrogenase in synthetic processes. Advanced Synthesis and Catalysis. 2013;355;1709-1714. DOI: https://doi.org/10.1002/adsc.201200959

[52] Johannes TW, Woodyer RD, Zhao H. Mint: Efficient regeneration of NADPH using an engineered phosphite dehydrogenase. Biotechnology and Bioengineering. 2007;96(1);18-26. DOI: https://doi.org/10.1002/bit.21168

[53] Petschacher B, Staunig N,

oxidase 2 for NAD(P)+

DOI: 10.1039/C0GC00595A

[55] Monti D, Ottolina G, Carrea G, Riva S. Mint: Redox reactions catalyzed by isolated enzymes. Chemical reviews,

Müller M, Schürmann M, Mink D, De Wildeman S, Gruber K, Glieder A. Mint: Cofactor specificity engineering of Streptococcus mutans NADH

biocatalytic oxidations. Computational and Structural Biotechnology Journal. 2014;9(14);e201402005. DOI: https:// doi.org/10.5936/csbj.201402005

[54] Hollmann F, Arends IW, Buehler K, Schallmey A, Bühler B. Mint: Enzymemediated oxidations for the chemist. Green Chemistry. 2011;13(2);226-265.

regeneration in

Zhao H. Mint: Relaxing the

org/10.1021/ja00299a044

**28**

2011;111(7);4073-4087. DOI: https://doi. org/10.1021/cr200111v

[63] Dong J, Fernández-Fueyo E, Hollmann F, Paul CE, Pesic M, Schmidt S, Wang Y, Younes S, Zhang W. Mint: Biocatalytic oxidation reactions: A chemist's perspective. Angewandte Chemie International Edition. 2018; 57(30);9238-9261. DOI: https://doi. org/10.1002/anie.201800343

[64] Mifsud M, Gargiulo S, Iborra S, Arends IW, Hollmann F, Corma A. Mint: Photobiocatalytic chemistry of oxidoreductases using water as the electron donor. Nature Communications. 2014; 5(1);1-6. DOI: 10.1038/ncomms4145

[65] Paul PEV, Sangeetha V, Deepika RG. Emerging trends in the industrial production of chemical products by microorganisms. In: Recent developments in applied microbiology and biochemistry. Academic Press. 2019. p. 107-125. DOI: https://doi.org/10.1016/ B978-0-12-816328-3.00009-X

[66] Wolberg M, Hummel W, Müller M. Mint: Biocatalytic reduction of β, δ-diketo esters: A highly stereoselective approach to all four stereoisomers of a chlorinated β, δ-dihydroxy hexanoate. Chemistry–A European Journal. 2001;5;7(21);4562-71. DOI: https://doi.org/10.1002/1521- 3765(20011105)7:21<4562::AID-CHEM4562>3.0.CO;2-4

[67] Xu F. in The Encyclopedia of Bioprocessing Technology: Fermentation, Biocatalysis,and Bioseparation, eds. Flickinger MC, and Drew SW. 1545-1554 (John Wiley & Sons,New York, 1999).

[68] Ose T, Watanabe K, Mie T, Honma M, Watanabe H, Yao M, Oikawa H, Tanaka I. Mint: Insight into a natural Diels–Alder reaction from the structure of macrophomate synthase. Nature. 2003;422(6928);185-9. DOI: 10.1038/nature01454

[69] Xu F. Mint: Applications of oxidoreductases: recent progress. Industrial Biotechnology. 2005; 1;1(1);38-50. DOI: https://doi. org/10.1089/ind.2005.1.38

[70] Danielsen S, Christensen BE. PCT patent WO2003047351-A(2003).

[71] Miller DR, Tizard IR, Keeton JT, Prochaska JF. PCT patent WO200135882-A(2001).

[72] Ensley BD, inventor; Matrix Design, assignee. Wound healing compositions and methods using tropoelastin and lysyl oxidase. United States patent US 6,808,707. 2004 Oct 26. US patent US6808707-B2(2004).

[73] Perrier E, Cenizo V, Bouez C, Sommer P, Damour O, Gleyzal C, Andre V, Reymermier C, inventors; Centre National de la Recherche Scientifique CNRS, Coletica, assignee. Stimulation of the synthesis of the activity of an isoform of lysyl oxidase-like LOXL for stimulating the formation of elastic fibres. United States patent application US 10/852,065. 2004 Dec 16. US patent US2004253220-A1(2004).

[74] Szynol A, De Soet JJ, Siebenvan Tuyl E, Bos JW, Frenken LG. Mint: Bactericidal effects of a fusion protein of llama heavy-chain antibodies coupled to glucose oxidase on oral bacteria. Antimicrobial Agents and Chemotherapy. 2004;;48(9):3390-3395.

[75] Novozymes AS. Danish patent DK200100630-A (2001).

[76] Wang HX, Ng TB. Mint: Purification of a novel low-molecularmass laccase with HIV-1 reverse transcriptase inhibitory activity from the mushroom Tricholoma giganteum. Biochemical and Biophysical Research Communications. 2004;315(2);450-4. DOI: 10.1016/j.bbrc.2004.01.064

**31**

**Chapter 3**

**Abstract**

Endothelium

*and Md. Ruhul Abid*

recovery of cardiac function.

fatty acid oxidation

**1. Introduction**

Role of Subcellular ROS in

Providing Resilience to Vascular

For decades, elevated levels of reactive oxygen species (ROS) have been associated

with the pathogenesis of cardiovascular diseases (CVD), including myocardial ischemia and infarction (MI). However, several large clinical trials failed to demonstrate beneficial outcomes in response to the global reduction of ROS in patients with underlying CVD. Recent studies from our and other labs showed that it is rather a critical balance between mitochondrial and cytosolic ROS than total ROS levels which determines resilience of coronary endothelial cells (EC). Here, we will discuss published and unpublished work that has helped elucidate the molecular mechanisms by which subcellular ROS levels, duration and localization modulate metabolic pathways, including glycolysis and oxidative phosphorylation, energy production and utilization, and dNTP synthesis in EC. These redox-regulated processes play critical roles in providing resilience to EC which in turn help protect existing coronary vessels and induce coronary angiogenesis to improve post-MI

**Keywords:** endothelial cell metabolism, angiogenesis, vascular endothelial growth factor (VEGF), nitric oxide, reactive oxygen species (ROS), glycolysis, dNTP,

A single layer of endothelial cells (ECs) that covers the vascular lumen and plexus exhibits great plasticity to adapt to environmental cues [1, 2]. It is fascinating how the vascular system, the largest organ system of the body, connects all organs to secure adequate nutrients and blood supply. For that reason, maintaining vascular homeostasis is crucial for the health of the cardiovascular system. In a healthy body, although the ECs are an intricate, dynamic system, they appear to be in a quiescent state [1]. In pathological conditions such as ischemia and infarction, ECs rapidly switch phenotype to form new vessels in a process known as sprouting angiogenesis [3]. Reactive oxygen species (ROS) are believed to play crucial roles in determining the phenotype and fate of EC in both physiological and pathological conditions. Recent work has shown that a critical balance between mitochondrial and cytosolic ROS levels, but not global ROS levels, modulates endothelial function, EC metabolism and angiogenesis, and thus determines resilience of coronary EC [4–8].

*Sarah R. Aldosari, Maan A. Awad, Frank W. Sellke* 

#### **Chapter 3**
