**2.2 Anti-inflammatory agents**

Anti-inflammatory properties of terpenoids were already described in various reviews [73, 74] but they were not investigated in cited papers so we did not discuss obtained results. It is worth mentioning that each extract obtained after SSF contained at least one compound that could demonstrate such activity: lactones which were produced by *Trichoderma viride* EMCC-107 [28]; limonene-1,2-diol, α-terpineol, (−)-carvone, α-tocopherol produced by *Diaporthe* sp. KY113119 [25]; 1-terpineol, L-linalool produced by *Antrodia camphorata* [30]; betulinic acid – *Inonotus obliquus* and [23]; α-pinene produced by *Saccharomyces cerevisiae* AXAZ-1 and *Kluyveromyces marxianus* IMB3 [27].

In the case of polyphenols we took the same approach – we only summarised the research that investigated anti-inflammatory properties of extracts obtained after SSF. When grape pomace was treated with *Trametes versicolor* TV-6 the concentration of phenolic acids, flavan-3-ols and rutin increased while the concentration of anthocyanins decreased. Those changes resulted in enhanced anti-inflammatory activity of obtained extracts which was measured by the inhibition of 5-lipoxygenase and hyaluronidase [20]. Also polyphenols produced by *Xylaria nigripes* demonstrated enhanced anti-inflammatory properties which were verified by the inhibition of cyclooxygenase-2 [24]. Additionally, in both cases, obtained extracts demonstrated neuroprotective properties. Studies carried out by Yin et al. [75] demonstrated that *A. niger* was able to release ferulic acid bound to various polysaccharides in the wheat bran. Obtained substances exhibited stronger anti-inflammatory activity than that of free ferulic acid which probably took place due to the presence of accompanying compounds in obtained extracts. Moreover, those released compounds could significantly inhibit intracellular malondialdehyde formation and the LPS-induced inflammation. It is difficult to assess which extracts demonstrated greater activity because authors of all abovementioned studies used different way to express results – the provided IC50 values or % of inhibition.

In the majority of cited papers anti-inflammatory properties of extracts containing polyphenols were not verified despite the fact that there were some compounds among them which demonstrate such activity [76, 77]. Research mainly focused on antioxidant properties [20, 22, 23, 31]. This applies to various studies involving organisms providing increase of particular phenolic compounds: *Rhizopus oligosporus* NRRL 2710 – rutin [22], *Prunus armeniaca* L – cinnamic acids and selected flavonols [31]; *Lactobacillus plantarum* CECT 748 – hydroxybenzoic acids and flavonols [78]; *Eurotium cristatum* YL-1 – isoflavones (daidzin, daidzein, genistin and genistein) [34]; *Rhizopus oryzae* and *Aspergillus foetidus* – tannins [35]; *Rhizopus oryzae* RCK2012 – phenolic acids [36]; *Aspergillus oryzae* LBA01 – 3,4-di- hydroxybenzoic acid, ferulic acid, vanillic acid and quercetin [37].

#### **2.3 Immunosuppressants**

Immunosuppressants could be obtained by SSF as well. It seems that one of the most common substance which was detected in studies involving SSF is mycophenolic acid (MA, **Table 1**). It works as a blocker in producing precursors for the synthesis of RNA and DNA, so as the result it blocks proliferative response of T and B lymphocytes [79]. In the research that involved SSF, authors did not verify properties of the obtained MA.

Another substance belonging to that group, classified as calcineurin inhibitor [80], is cyclosporin A (CA). This substance is used not only in transplant patients but also in treatment of glomerular disease. CA prevents calcineurin-dependent transcription in activated T cells. Based on studies that were aiming to produce CA we concluded that authors did not verify properties or safety of obtained substances but they focused on various methods for its extraction and purification. They mostly used butyl acetate [44, 48] or ethyl acetate [47]. Tacrolimus could be also produced by SSF by *Streptococcus hygroscopicus*. The whole process of bacterial cultivation and the extraction of that compounds was covered by the patent [49].

#### **2.4 Anticoagulants**

Another group of substances that could be produced by SSF is anticoagulants. Blood coagulation is a physiological process, which consists of a series of coagulation factors and proteolytic activation steps, which lead to the production of

**337**

*The Application of Solid State Fermentation for Obtaining Substances Useful in Healthcare*

thrombin – the main coagulation enzyme. The majority of research was focused on fibrinolytic enzymes (FE, **Table 1**) – subtilisin, however, in one case it was nattokinase [55]. In the majority of cited papers authors verified anticoagulant properties of obtained enzymes and they used different methods – some of them applied spectrophotometric method which measured the increase of turbidity at 275 nm caused by added enzyme [53, 54, 58, 60, 62, 65] and others measured zones of clearance in

Another compound which could be considered as a putative anticoagulant is halotolerant chitinase. Its properties were confirmed [50, 51] by testing how this enzyme could dissolve fibrin in time. Moreover, Meruvu et al. [51] showed that it

Terpenoids are well-known for their cytotoxic activity and there were various studies investigating such activity against cancer cells. Anticancer mechanism of terpene or essential oils that contain them was described in various review papers [81–83]. Several compounds belonging to that class could be obtained by SSF (**Table 1**): limonene from orange waste [25]; linalool, geraniol and β-caryophyllene from millet [28, 30]; andrastin A and C on malt extract agar [70]. Properties of extracts that contained those and other terpenoids were not investigated in cited

Another group of bioactive compounds which was shown to hold anticancer activity is polyphenols [84, 85]. The following substances occurred in cited reviews and studies that we summarised in **Table 1**: cinnamic acids [22], daidzein and genistein [86], quercetin [35, 87], and tannins [34]. Since authors of cited research papers (**Table 1**) did not test obtained extracts against those properties, we did not

In the study of Cho et al. (2019) authors demonstrated that fatty acids detected in extracts obtained from silkworm larvae powder fermented by *Aspergillus kawachii* demonstrated such activity against human hepatocellular carcinoma [68]. It was shown that fermentation increased concentration of those compounds, especially oleic and linoleic acids. This phenomenon took place due to the enhancement of cell apoptosis and suppression of protein responsible for preventing the apoptosis. The value of that research is particularly significant because so far, polyunsaturated fatty acids (PUFAs) attracted most attention in the context of colorectal cancer [87].

**3. Groups of microorganisms demonstrating greatest potential for the** 

It was demonstrated that the representatives of the *Bacillus* genus were able to produce fibrinolytic enzymes by SSF [52–58, 60]. That ability is mostly assigned to the expression of *fibE* gene which encodes enzyme called subtilisin [88] which solubilises blood clots. Gene expression was not investigated in cited papers so the ability to produce those enzymes by tested strains could be the result of other genes

*Bacillus* genus is successfully used in solid-state fermentation to improve antimicrobial activity of fermented food. Rochín-Medina et al. [19] who tried to determine

optimal bioprocessing conditions for SSF of spent coffee grounds by *Bacillus clausii* achieved increase of flavonoid and total phenolic contents by 13 and 36%,

**production of health-promoting properties in SSF**

papers so we decided not to discuss that aspect in the current chapter.

*DOI: http://dx.doi.org/10.5772/intechopen.94296*

solid media containing fibrinogen [64, 66, 67].

held antifungal activity.

**2.5 Anticancer agents**

discuss that phenomenon.

**3.1** *Bacillus* **genus**

expression.

*The Application of Solid State Fermentation for Obtaining Substances Useful in Healthcare DOI: http://dx.doi.org/10.5772/intechopen.94296*

thrombin – the main coagulation enzyme. The majority of research was focused on fibrinolytic enzymes (FE, **Table 1**) – subtilisin, however, in one case it was nattokinase [55]. In the majority of cited papers authors verified anticoagulant properties of obtained enzymes and they used different methods – some of them applied spectrophotometric method which measured the increase of turbidity at 275 nm caused by added enzyme [53, 54, 58, 60, 62, 65] and others measured zones of clearance in solid media containing fibrinogen [64, 66, 67].

Another compound which could be considered as a putative anticoagulant is halotolerant chitinase. Its properties were confirmed [50, 51] by testing how this enzyme could dissolve fibrin in time. Moreover, Meruvu et al. [51] showed that it held antifungal activity.

#### **2.5 Anticancer agents**

*Biotechnological Applications of Biomass*

In the case of polyphenols we took the same approach – we only summarised the research that investigated anti-inflammatory properties of extracts obtained after SSF. When grape pomace was treated with *Trametes versicolor* TV-6 the concentration of phenolic acids, flavan-3-ols and rutin increased while the concentration of anthocyanins decreased. Those changes resulted in enhanced anti-inflammatory activity of obtained extracts which was measured by the inhibition of 5-lipoxygenase and hyaluronidase [20]. Also polyphenols produced by *Xylaria nigripes* demonstrated enhanced anti-inflammatory properties which were verified by the inhibition of cyclooxygenase-2 [24]. Additionally, in both cases, obtained extracts demonstrated neuroprotective properties. Studies carried out by Yin et al. [75] demonstrated that *A. niger* was able to release ferulic acid bound to various polysaccharides in the wheat bran. Obtained substances exhibited stronger anti-inflammatory activity than that of free ferulic acid which probably took place due to the presence of accompanying compounds in obtained extracts. Moreover, those released

compounds could significantly inhibit intracellular malondialdehyde formation and the LPS-induced inflammation. It is difficult to assess which extracts demonstrated greater activity because authors of all abovementioned studies used different way to

In the majority of cited papers anti-inflammatory properties of extracts containing polyphenols were not verified despite the fact that there were some compounds among them which demonstrate such activity [76, 77]. Research mainly focused on antioxidant properties [20, 22, 23, 31]. This applies to various studies involving organisms providing increase of particular phenolic compounds: *Rhizopus oligosporus* NRRL 2710 – rutin [22], *Prunus armeniaca* L – cinnamic acids and selected flavonols [31]; *Lactobacillus plantarum* CECT 748 – hydroxybenzoic acids and flavonols [78]; *Eurotium cristatum* YL-1 – isoflavones (daidzin, daidzein, genistin and genistein) [34]; *Rhizopus oryzae* and *Aspergillus foetidus* – tannins [35]; *Rhizopus oryzae* RCK2012 – phenolic acids [36]; *Aspergillus oryzae* LBA01 – 3,4-di- hydroxy-

Immunosuppressants could be obtained by SSF as well. It seems that one of the most common substance which was detected in studies involving SSF is mycophenolic acid (MA, **Table 1**). It works as a blocker in producing precursors for the synthesis of RNA and DNA, so as the result it blocks proliferative response of T and B lymphocytes [79]. In the research that involved SSF, authors did not verify

Another substance belonging to that group, classified as calcineurin inhibitor [80], is cyclosporin A (CA). This substance is used not only in transplant patients but also in treatment of glomerular disease. CA prevents calcineurin-dependent transcription in activated T cells. Based on studies that were aiming to produce CA we concluded that authors did not verify properties or safety of obtained substances but they focused on various methods for its extraction and purification. They mostly used butyl acetate [44, 48] or ethyl acetate [47]. Tacrolimus could be also produced by SSF by *Streptococcus hygroscopicus*. The whole process of bacterial cultivation and the extraction of that compounds was covered by the patent [49].

Another group of substances that could be produced by SSF is anticoagulants. Blood coagulation is a physiological process, which consists of a series of coagulation factors and proteolytic activation steps, which lead to the production of

express results – the provided IC50 values or % of inhibition.

benzoic acid, ferulic acid, vanillic acid and quercetin [37].

**2.3 Immunosuppressants**

properties of the obtained MA.

**2.4 Anticoagulants**

**336**

Terpenoids are well-known for their cytotoxic activity and there were various studies investigating such activity against cancer cells. Anticancer mechanism of terpene or essential oils that contain them was described in various review papers [81–83]. Several compounds belonging to that class could be obtained by SSF (**Table 1**): limonene from orange waste [25]; linalool, geraniol and β-caryophyllene from millet [28, 30]; andrastin A and C on malt extract agar [70]. Properties of extracts that contained those and other terpenoids were not investigated in cited papers so we decided not to discuss that aspect in the current chapter.

Another group of bioactive compounds which was shown to hold anticancer activity is polyphenols [84, 85]. The following substances occurred in cited reviews and studies that we summarised in **Table 1**: cinnamic acids [22], daidzein and genistein [86], quercetin [35, 87], and tannins [34]. Since authors of cited research papers (**Table 1**) did not test obtained extracts against those properties, we did not discuss that phenomenon.

In the study of Cho et al. (2019) authors demonstrated that fatty acids detected in extracts obtained from silkworm larvae powder fermented by *Aspergillus kawachii* demonstrated such activity against human hepatocellular carcinoma [68]. It was shown that fermentation increased concentration of those compounds, especially oleic and linoleic acids. This phenomenon took place due to the enhancement of cell apoptosis and suppression of protein responsible for preventing the apoptosis. The value of that research is particularly significant because so far, polyunsaturated fatty acids (PUFAs) attracted most attention in the context of colorectal cancer [87].
