**2. Methods**

*Lactobacillus* and *Bifidobacterium* strains were from the collection of microorganisms of the normal microflora of *G. N. Gabrichevsky* Institute for Epidemiology and Microbiology, and probiotics *Bifidin* and *Acilact* were products of our institute. Bacteria were grown in media containing casein hydrolysate and yeast autolysate. LSSM were extracted from the protein fractions 27–220 kD using isoelectrofocusing (IEF) in a plate of polyacrylamide gel (PAG) in a gradient of pH 4–8. Identification of proteins was performed by electroblotting on a hydrophobic membrane and staining with SYPRO blot stain (Bio-Rad Lab.). Nonstained proteins were evaluated by other spectrophotometric methods. The distribution of LSSM among proteins was determined by treatment of blots with biotinylated GC (GC-b) containing multiple residues of sugar(s) linked to the polyacrylamide (PA) linear core (like in external phenotypes of mucins and mucin-type glycans) (www.lectinity.com;


**73**

by human blood leukocytes [9, 17].

*Metabolite Multiprobiotic Formulas for Microbial Health DOI: http://dx.doi.org/10.5772/intechopen.86449*

optical density at 420 nm.

**3. Results and discussion**

**3.1 Symbiotic lectins as system regulators and delivery agents**

LSSM function as metabolomebiotics regulating metabolome according to the principle "LSSM network—whole organism metabolome network or interactome" [15]. The network of LSSM is created in the following manner: lectin molecule of determined molecular weight (in the *Laemmli* system) is represented by LS including forms of varying charge and possessing a range of biological and physiological activities; such a minimal LS can be further transformed in a natural manner into extended network of complexes and supramolecular ensembles as a result of directional and sequential cascade binding of carbohydrates and GC. As a result of forming complexes and ensembles, lectin specificity of complexes and ensembles can be modified or changed during further development of recognition cascade network that will change the summary vector of specificity of LS. The latter will result in dynamic qualitative and quantitative changes of the local biotope surrounding. Thus, the final whole resulting network of LSSM (as metabolomebiotics) will regulate the whole metabolome and interactome of organism involving human glycome (carbohydrates and GC: glycoproteins, glycoenzymes, glycolipids, receptors, and others [16]). The metabolome possesses the ability to back direct and reversible effects of LSSM representing a part of hierarchic interactome. Multiple forms of adapted functioning LSSM microbiocenosis in the biotope will depend on the originality developed in a joint process of evolution involving host body local infrastructures for the distribution and disposition of microbiocenoses (organ-type constructions of both host and microbiocenosis interests are possible) [11, 12]. LSSM are ready to realize biologically active GC (as prebiotics, therapeutic agents, and metabiotics) in such symbiotic organs. The network of LSSM functions as noncellular simulators of symbiotics (probiotics) in the direct or indirect (through human higher hierarchic protection systems) predictable manners. For example, of communications between LSSM and own human protection systems, LSSM (as well as phytohemagglutinins from plants of medical significance) and artificial polymeric GC influenced peritoneal macrophage mobility in a coupled manner depending on doses of agents; LSSM were involved in modulation of cytokine production

New useful properties of LSSM can be predicted and verified (cofunctioning to enzymes, adhesion, etc.), based on the fact that LSSM form a functional

**Table 1**) followed by final treatment with streptavidin-peroxidase conjugate. The bound peroxidase on the blot was registered in the presence of a chemiluminescent substrate in regime of a real time in the system BioChemi System (UVP, CA). Antimicrobial activities and synergism of LSSM, antibiotics, and phytolectins were tested on solid agar media during the prolonged growth and survival of communicative fungal bodies (CFB) in the presence of disks of antimicrobials. Biosurfactants were tested and calculated using detection of sample drop activity against mineral oil film on water surface (the appearance of transparent circles). Amino acid compositions of samples were established using standard amino acid analyzer column chromatography. Oxidoreductase systems were detected on blots after IEF-PAG, resulting in kinetic protein stain disappearance (decolorization). Hydrolase systems were visually evaluated on blots after IEF-PAG (resulting in hydrolysis, splitting, and partial asymmetrical disappearance of protein bands). *Maillard* reaction products were partially evaluated as browning in culture supernatant according to

**Table 1.** *A list of synthetic GC used.* *Metabolite Multiprobiotic Formulas for Microbial Health DOI: http://dx.doi.org/10.5772/intechopen.86449*

*Oral Health by Using Probiotic Products*

**2. Methods**

and antibiotics) (see **Table 2**) [8, 9, 13, 14].

with enzymes of all known classes; and agents possessing antipathogenic synergism of different LS in antimicrobial combinations (between LS of *Lactobacillus* species and *Bifidobacterium* species, between genera *Lactobacillus* and *Bifidobacterium*, between LS of probiotic bacteria and lectins from medical plants, between LSSM

LS reveal significantly higher multifunctionality (antimicrobial, cytokine-like, others) and adaptive ability in surroundings in comparison to any component of LS. Applied prospects of LSSM in microbial associations of biotopes in the human body are of promised interest. LSSM and their reactive GC support balanced functioning in organism in respect to evolutionary created mucosal organ-like infrastructures of mutual interest for human and biotope microbiocenoses (MB) [12]. The purpose of the review is to evaluate our approaches in creation of probiotic

metabolite compositions influencing and improving health of human biotope microbiocenoses. The data presented will be of interest for investigators in the fields of both medical microbiology and laboratory and industrial medical biotechnology.

*Lactobacillus* and *Bifidobacterium* strains were from the collection of microorganisms of the normal microflora of *G. N. Gabrichevsky* Institute for Epidemiology and Microbiology, and probiotics *Bifidin* and *Acilact* were products of our institute. Bacteria were grown in media containing casein hydrolysate and yeast autolysate. LSSM were extracted from the protein fractions 27–220 kD using isoelectrofocusing (IEF) in a plate of polyacrylamide gel (PAG) in a gradient of pH 4–8. Identification of proteins was performed by electroblotting on a hydrophobic membrane and staining with SYPRO blot stain (Bio-Rad Lab.). Nonstained proteins were evaluated by other spectrophotometric methods. The distribution of LSSM among proteins was determined by treatment of blots with biotinylated GC (GC-b) containing multiple residues of sugar(s) linked to the polyacrylamide (PA) linear core (like in external phenotypes of mucins and mucin-type glycans) (www.lectinity.com;

1. α/β-D-GalNAc-PA-b (animal mucins, T antigens)\*

3. β-D-GlcNAc-PA-b (insect chitins and chitosans) 4. β-D-Gal-PA-b (plant or animal galactans) 5. β-D-Gal-3-sulfate-PA-b (acidic animal galactans)

7. α-D-Man-6P-PA-b (yeast phosphomannans) 8. α-L-Fuc-PA-b (fucans from brown algae) 9. α-L-Rha-PA-b (Gram-negative rhamnans) 10. MDP-PA-b (bacterial peptodoglycans)

11. Adi-PA-b (A-blood group substance GalNAcβ1-3GalβА1-) 12. Fs-PA-b (Forssman antigen GalNAcβ1-3GalNAcβА1-) 13. Tαα-PA-b (bacterial antigen Galα1-3GalNAcα1-)

2. β-D-GalNAc-PA-b (animal mucins)

6. α-D-Man-PA-b (yeast mannans)

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**Table 1.**

*A list of synthetic GC used.*

*\*In brackets—natural substances which are imitated*

**Table 1**) followed by final treatment with streptavidin-peroxidase conjugate. The bound peroxidase on the blot was registered in the presence of a chemiluminescent substrate in regime of a real time in the system BioChemi System (UVP, CA). Antimicrobial activities and synergism of LSSM, antibiotics, and phytolectins were tested on solid agar media during the prolonged growth and survival of communicative fungal bodies (CFB) in the presence of disks of antimicrobials. Biosurfactants were tested and calculated using detection of sample drop activity against mineral oil film on water surface (the appearance of transparent circles). Amino acid compositions of samples were established using standard amino acid analyzer column chromatography. Oxidoreductase systems were detected on blots after IEF-PAG, resulting in kinetic protein stain disappearance (decolorization). Hydrolase systems were visually evaluated on blots after IEF-PAG (resulting in hydrolysis, splitting, and partial asymmetrical disappearance of protein bands). *Maillard* reaction products were partially evaluated as browning in culture supernatant according to optical density at 420 nm.
