Production of D S-threo-isocitric acid (ICA) by yeast meets serious difficulties since it is accompanied by a simultaneous production of citric acid (CA) in significant amounts that reduces the yield of desired product. In order to develop an effective process of ICA production, 60 yeast strains of different genera (Candida, Pichia, Saccharomyces, Torulopsis, and Yarrowia) were tested for their ability to produce ICA from rapeseed oil; as a result, wild-type strain Yarrowia lipolytica VKM Y-2373 and its mutant Y. lipolytica 704-UV4-A/NG50 were selected as promising ICA producers. The effects of temperature, pH, aeration, and concentrations of rapeseed oil, iron, and itaconic acid on ICA production by selected strains were studied. Under optimal conditions (pH 6.0; aeration 50-55 %; rapeseed oil concentration of 20-60 gl(-1), iron ion concentration of 1.2 mg l(-1), and itaconic acid amount of 30 mM), selected strains of Y. lipolytica produced predominantly ICA with a low amount of a by-product, CA.
The basic tendency in the field of plant protection concerns with reducing the use of pesticides and their replacement by environmentally acceptable biological preparations. The most promising approach to plant protection is application of microbial metabolites. In the last years, bactericidal, fungicidal, and nematodocidal activities were revealed for citric, succinic, α-ketoglutaric, palmitoleic, and other organic acids. It was shown that application of carboxylic acids resulted in acceleration of plant development and the yield increase. Of special interest is the use of arachidonic acid in very low concentrations as an inductor (elicitor) of protective functions in plants. The bottleneck in practical applications of these simple, nontoxic, and moderately priced preparations is the absence of industrial production of the mentioned organic acids of required quality since even small contaminations of synthetic preparations decrease their quality and make them dangerous for ecology and toxic for plants, animals, and human. This review gives a general conception on the use of organic acids for plant protection against the most dangerous pathogens and pests, as well as focuses on microbiological processes for production of these microbial metabolites of high quality from available, inexpensive, and renewable substrates.
Arachidonic acid (AA) has a wide range of applications in medicine, pharmacology, diet, infant nutrition, and agriculture, due to its unique biological properties. The microbiological processes involved in AA production usually require carbohydrate substrates. In this paper, we propose a method for AA production from glycerol, an inexpensive and renewable carbon substrate that is produced by the fungal strain, Mortierella alpina NRRL-A-10995. Our experimental results showed that the optimum pH values required for fungal growth and the production of lipids and AA were different and depended on the growth phase of the fungus. The AA production was shown to be extremely sensitive to acidic pH values and was completely inhibited at a pH of 3.0. The optimum temperature for AA production was 20-22 • C. Continuous cultivation of M. alpina occurred in a glycerol-containing medium, and growth limitations were implemented through the addition of nitrogen and the selection of optimal conditions (pH 6.0, 20 • C). This ensured that active AA production occurred (25.2% of lipids and 3.1% of biomass), with the product yield from the consumed glycerol being 1.6% by mass and 3.4% by energy.
The process of succinic acid (SA) production represents the combination of microbial synthesis of α-ketoglutaric acid from rapeseed oil by yeast Yarrowia lipolytica VKM Y-2412 and subsequent decarboxylation of α-ketoglutaric acid by hydrogen peroxide to SA that leads to the production of 69.0 g l(-1) of SA and 1.36 g l(-1) of acetic acid. SA was isolated from the culture broth filtrate in a crystalline form. The SA recovery from the culture filtrate has certain difficulties due to the presence of residual triglycerides of rapeseed oil. The effect of different methods of the culture filtrate treatment and various sorption materials on the coagulation of triglycerides was studied, and as a result, the precipitation of residual triglycerides by acetone was chosen. The subsequent isolation procedures involved the decomposition of H2O2 in the filtrate followed by filtrate bleaching and acidification with a mineral acid, evaporation of filtrate, and SA extraction with ethanol from the residue. The purity of crystalline SA isolated from the culture broth filtrate achieved 97.6-100 %. The product yield varied from 62.6 to 71.6 % depending on the acidity of the supernatant.
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