


Vol 53, No 8 (2017)
- Year: 2017
- Articles: 7
- URL: https://journals.rcsi.science/0003-6838/issue/view/9110
Producers, Biology, Selection, and Gene Engineering
Mutations in the fusA Gene Encoding Elongation Factor G in the Coryneform Bacterium Lead to Increased Lysine Production
Abstract
Resistance to fusidic acid in Corynebacterium glutamicum and Brevibacterium flavum is associated with mutations in the fusA gene, which encodes the elongation factor G (EF-G). Two to ten percent of fusidic acid-resistant clones were shown to produce more lysine than parent strains. Sequencing of the fusA gene in clones with a high level of lysine production made it possible to find two mutations in the gene at position 1383—С1383G and С1383А. These mutations cause amino acid replacement at position 461 in the protein EF-G, namely, histidine is substituted by glutamine (H461Q). The mutation С1383G was introduced in the chromosomal copy of the fusA gene in C. glutamicum and B. flavum strains by homologous recombination. All clones containing the mutant variant of the fusA gene produced 10% more lysine than the parent strains.



Bacterial Strain Alcaligenes denitrificans C-32 Containing Two Nitrilases with Different Substrate Specificities
Abstract
Two genes encoding nitrilases with different properties have been found in an Alcaligenes denitrificans C-32 strain with high nitrilase activity that is currently used as a biocatalyst for commercial ammonium acrylate production. Both genes were expressed in E. coli, and the properties of the recombinant nitrilases were studied. One of these genes, which is designated as nitC1, controlled the formation of nitrilase that preferred aliphatic nitriles (acrylonitrile and butyronitrile) as best substrates. The nucleotide sequence of the gene nitC1 was almost (99%) identical to the gene sequence of an aliphatic nitrilase from Acidovorax facilis 72W (DQ4444267). In turn, nitC2 had a high level of homology (85%) with the arylacetonitrilase gene from Alcaligenes faecalis JM3 (D13419). Benzyl cyanide was shown to be the best substrate for nitC2-encoded nitrilase. In light of the results of DNA homology and differences in substrate specificity, the NitC2 and NitC1 nitrilases from Alcaligenes denitrificans C-32 were allocated to the groups of aliphatic nitrilases and arylacetonitrilases, respectively.



Development of a New Amycolatopsis orientalis Strain for High-Yield Eremomycin Production
Abstract
A multistep random UV mutagenesis of an eremomycin-producing strain Amycolatopsis orientalis VKPM Ac-1125 with rather low productivity (0.5 g/L) and the further selection of the most high-yield mutant strains have resulted in a highly productive E 13-26 strain. After optimization of the fermentation medium by multifactor analysis, the most efficient composition for the fermentation medium was established, %: glycerol, 8; soybean meals, 2.0; MgCl2, 0.02; KH2PO4, 0.02; KNO3, 0.6; CaCl2, 0.3; skim milk powder, 0.46; and glucose, 0.8. The maximum productivity of this strain on the optimized medium reached 3.42 ± 0.17 g/L. Therefore, the obtained strain is considered to be very promising for further use in the selection and development of technology for the large-scale eremomycin production.



Obtaining and Study of Callus and Suspension Plant Cell Cultures of Tribulus terrestris L., a Producer of Steroidal Glycosides
Abstract
Callus and suspension plant cell cultures of Tribulus terrestris L., a valuable medicinal plant producing steroidal glycosides, were obtained. The seeds from an American population of T. terrestris were used as explants. Regulation of the production and growth of cell cultures, as well as the biosynthetic characteristics of the cell lines, were studied. The combination of phytohormones of 2,4-D (2.0 mg/L) and BAP (1.0 mg/L) was found to be optimal for callus induction and cultivation. Suspension cell culture obtained in liquid medium of the same composition showed such high growth characteristics during prolonged cultivation (more than 2 years) as a maximum accumulation of dry biomass of 13 g/L, specific growth rate at exponential phase of 0.24 day–1, and economical coefficient of 0.39. A semicontinuous mode of cultivation was used to grow the plant cell suspension in a lab-scale bioreactor. Screening of the steroidal glycosides in the obtained cell cultures was carried out. Steroidal glycosides were not found in the callus cultures. However, as was demonstrated by TLC and UPLC ESI MS methods, the suspension culture contained furostanol glycosides, and their amount increased during the cultivation process. These results support the hypothesis of the autoselection of cultivated cells containing compounds promoting their proliferation in vitro.



Calculation of Biodiesel Fuel Characteristics Based on the Fatty Acid Composition of the Lipids of Some Biotechnologically Important Microorganisms
Abstract
The interdependences between the structure of fatty acid and biofuel characteristics obtained from these fatty acids were briefly reviewed. The fatty acid compositions of the lipids of yeasts and phototrophic microorganisms were analyzed. The main parameters of the biodiesel (iodine value, cetane number, and kinematic viscosity) that can be made from the lipids of these microorganisms were calculated based on the data and compared to the current standards. The lipids of the yeast Rhodosporidium toruloides VKPM Y-3349 were shown to be the most suitable for biofuel production due to the composition and content of fatty acid. The possibilities of a decrease in the prime cost of microbial lipids (along with plant oils) that would make them competitive raw material for biofuel production were considered.



Biological Preparation Technology
Starch Phosphate Microgels for Controlled Release of Biomacromolecules
Abstract
Gel-forming starch phosphates with a content of acidic phosphate groups ranging from 2.1 to 3.8 mmol/g were obtained in a phosphoric acid-urea system. The rates of starch phosphate degradation in buffer solution in the absence of amylase and in the presence of this enzyme were investigated in vitro. Starch phosphate gels were shown to be prone to enzymatic degradation. However, the rate of biodegradation decreased gradually as the content of phosphate groups increased. The use of microgels with average particle sizes in the range of 7.8–60.1 μm for the production of controlled-release preparations of interferon-alpha 2b has been demonstrated to be possible in principle.



Preparation and Properties of Nanocolloidal Rhenium Sulfide Solution for Lymphoscintigraphic Methods of Micrometastase Examination
Abstract
A method for the preparation of a nanocolloidal solution of rhenium sulfide is proposed. It includes the following stages: interaction of ammonium perrhenate and sodium thiosulfate in an aqueous solution of gelatin in an acidic medium with heating to 70–80°C; neutralization to pH 7.0; dialysis against saline; desalination and concentrating by ultrafiltration. The resulting colloidal solution contains 85% of particles with an optimal size of 80–100 nm and can be used as a diagnostic tool for lymphoscintigraphy of sentinel lymph nodes in patients with cancer and other diseases.


