Vol 53, No 4 (2017)

This paper summarizes modern views on the physiological and biochemical properties of aerobic methylobacteria as producers of bioplastics and the prospects for their use in various areas of modern biotechnology.

The study was devoted to the synthesis of pentyl glucosides (PenG_n) and isopentyl glucosides (Iso-PenG_n) by transglycosylation using recombinant cyclodextrin glycosyltransferase from Bacillus circulans A11, β-cyclodextrin as a glucosyl donor and 1-pentanol and isopentanol as acceptors. TLC and MS analysis indicated at least 3 products which were in accordance with PenG_n and IsoPenG_n having glucose, maltose and maltotriose attached to the alkyl groups of both alcohols. Two products of each glucoside were purified by preparative TLC and their structures were identified by NMR technique to be pentyl-α-D-glucopyranoside (PenG₁), pentyl-α-D-maltopyranoside (PenG₂), isopentyl-α-D-glucopyranoside (IsoPenG1) and isopentyl- α-D-maltopyranoside (IsoPenG₂). The effect of water-in-hexadecane emulsion on emulsion-forming properties showed that PenG₂ had the highest emulsifying activity. Adding PenG₂ to the insoluble Corynebacterium glutamicum amylomaltase from Escherichia coli transformants (A406R), helped to perform it to more soluble conformation. Moreover, it was found that PenG_1,2 exhibited a higher antibacterial activity against E. coli ATCC 25922 than that of IsoPenG_1,2. Hence, the biological properties of the synthesized products may be useful for their applications as emulsifying, solubilizing and antibacterial agents.

The properties of two extracellular proteases of Aspergillus ochraceus VKM F-4104D micromycete with plasmin-like activity have been studied. It has been shown that the enzymes differ in pI (5.05 and 6.83) and have similar molecular weights (about 32 and 35 kDa), pH optima (pH 9.0–10.00 at 45°C), and specificities of action on a limited set of chromogenic peptide substrates of trypsin-like proteases. According to inhibitory analysis, both enzymes belong to the serine proteases. Their properties appeared to be similar to those of the protease, protein C activator, which is the main proteolytic enzyme of A. ochraceus VKM F-4104D. Most likely, proteases of this micromyсetes are isoenzymes.

The application potential of microfungal strains (Cadophora malorum, Mucor circinelloides, Trichoderma viride, nonsporulating culture Mycelia sterilia) as promising lipid producers is investigated. The C. malorum strain is found to be optimal for oil sludge recycling into biofuel. Its palmitic acid content is 52.9%, and it ensures a cetane number of the obtained biodiesel. The ability of the C. malorum strain to degrade n-alkanes and polyaromatic hydrocarbons allows the effective bioremediation of oil sludge to be performed.

Binary biofertilizers consisting of a rhizobial strain specific to the plant Rhizobium leguminosarum and a strain-producer of the RapA1 adhesin protein, which enhances the adhesion of rhizobacteria to the plant roots, have been designed. A system consisting of highly productive bacteria strains and additional strains that enhance the action of the main component have been obtained.

The possibility of analyzing bacterial cells infected by a specific bacteriophage, using Escherichia coli as an example, with an acoustic sensor directly in suspensions with different initial electrical conductivities was studied. The analysis was based on measurement of the time dependence of phase and the complete loss of output sensor signals of fixed frequency before and after biological interaction of microbial cells and bacteriophages. The aforementioned sensor makes it possible to detect bacterial cells and assess their viability in conducting suspensions. It was shown that the conductivity of the buffer solution should not exceed 10 μS/cm and the minimum detectable concentration of microbial cells was ~10⁴ cells/mL.

A polyelectrolyte-based enzymatic diagnosticum with a precipitation detection system that can be used as a biosensor was created. The detection method was based on the change in polyelectrolyte microcapsule weight with respect to the urea content. The possibility of biosensor reutilization was demonstrated. The appropriate ionic precipitator causing precipitation of insoluble carbonate within the microcapsules and the optimal microcapsule titre were found. In the solution of monovalent anions (chlorides), the activity of encapsulated urease was shown to increase monotonically as the square root of the ionic strength depending on the elevation of the salt content. The activity drastically increased in a narrow concentration interval (0.6–0.8 mM) of divalent anions (sulfates) and reached the level of the native enzyme activity.