42 results about "Torulopsis glabrata" patented technology

The invention relates to a method for adding alpha-ketoglutarate dehydrogenase inhibitor to realize excessive accumulation of alpha-ketoglutaric acid, which belongs to the technical field of the metabolic regulation optimized fermentation process of the protein level. The method of the invention comprises following steps: utilizing multi - vitamin - auxotrophic yeast of Torulopsis glabrata CCTCC M202019 as producing strains, regulating the activity of alpha-ketoglutarate dehydrogenase through adding the alpha-ketoglutarate dehydrogenase inhibitor: hydrogen peroxide, methotrexate, sodium hypochlorite or hydroxyamino in culture medium, purposively lowing the activity of the alpha-ketoglutarate dehydrogenase, reducing the degradation of the alpha-ketoglutaric acid in the metabolic process, and achieving the aim of the excessive accumulation of the alpha-ketoglutaric acid. The method of the invention cuts off carbon metabolism flow on a node of the alpha-ketoglutaric acid through regulating the stream distribution of carbon metabolism and the carbon metabolism flow, wherein the maximum output of the alpha-ketoglutaric acid reaches 23.2g / L, which is increased by 12.2% compared with the control. The invention provides a new thought for the fermentation research of TCA cycle intermediate metabolite.

A constructing method for non-resistance badge auxotrophy smoothing ball false yeast belongs to biological engineering technology field. The invention is a system method for breeding large fragment deletion non-resistance badge auxotrophy smoothing ball false yeast comprising obtaining large fragment deletion consanguinity recombination fragments by blending PCR method, preparing high efficient sprout fungi electric conversion competence, concentrating candio-hermal and screening restrictive culture medium. Goal auxotrophy bacterial strain can be obtained quickly within 3 to 5 days by a suit system method. The deletion of large fragments avoids efficiently reversion and consanguinity recombination of exogenous plasmid. Furthermore, the method can be used many times in a same leaving strain so that multiplex auxotrophy for following genetic engineering study can be obtained because any resistance badge is not used in entire process.

The invention discloses a method for increasing yield of pyruvic acid, belonging to the fermentation engineering field. The method is as follows: when Torulopsis glabrata (CCTCC NO: M202019) is used to ferment and produce pyruvic acid, aspartic acid is added to increase the acid stress resistance of Torulopsis glabrata. When the pH values are 5.5 and 4.5, the yields of pyruvic acid are separately55.8g / L and 30.4g / L which are separately increased by 12.4% (49.6g / L) and 48.4% (20.5g / L) than the control groups without aspartic acid; and the method has better technical effect and wide application prospect.

The invention relates to a torulopsis glabrata mutant strain and an application thereof in producing pyruvic acid through fermentation and provides a method for producing pyruvic acid through fermentation and culture of the strain, and the strain is the torulopsis glabrata mutant strain (CGMCC N0. 3077). The strain is a mutant strain that takes torulopsis glabrata NBRC 0005 as an original strain and is finally determined after going through ultraviolet mutation, preliminary screening of mutant strains and repeated screening by fermentation of a triangular flask. Under optimized medium component and fermentation conditions, the mutant strain takes glucose as the raw material to be fermented in a fermenter and produce over 0.5M of pyruvic acid. Counted by pyruvic acid sodium, the pyruvic acid reaches more than 5.5 percent, the conversion rate of saccharic acid is more than 30 percent and the fermentation period ranges from 45-55h. The technology is characterized by low content of competitive byproducts of pyruvic acid during downstream extraction process and simplified extraction process.

The invention discloses a method for microbial fermentative synthesis of α-ketoglutaric acid, in which calcium carbonate is added to a culture medium and the concentration of biotin is increased to promote the generation of a large amount of α-ketoglutarate during pyruvate fermentation. During the fermentative production of pyruvate by Torulopsis glabrata CCTCC M202019, delaying the addition of calcium carbonate inhibited the formation of α-KG and increased the carbon molar ratio of pyruvate to α-KG (C PYR / C KG ), and increasing the concentration of calcium carbonate in the medium will promote the accumulation of a large amount of α-KG, and when the concentration of calcium carbonate is 40g / L, it is most conducive to the formation of α-KG. Keep the concentration of calcium carbonate in the medium constant, increase the concentration of biotin in the medium, promote the concentration of α-KG to rise continuously and C PYR / C KG The value decreased continuously, and the accumulation of α-KG was 23.5g / L when the biotin concentration was 60μg / L. When there is Ca 2+ When present, the activity of intracellular pyruvate carboxylase can be increased by 40%, while the activity of pyruvate dehydrogenase system does not change significantly. Ca in medium 2+ The increase of the concentration of biotin and biotin can significantly increase the activity of pyruvate carboxylase, so that T. glabrata shifts from fermentative production of pyruvate to synthesis of high-concentration α-KG.

A constructing method for non-resistance badge auxotrophy smoothing ball false yeast belongs to biological engineering technology field. The invention is a system method for breeding large fragment deletion non-resistance badge auxotrophy smoothing ball false yeast comprising obtaining large fragment deletion consanguinity recombination fragments by blending PCR method, preparing high efficient sprout fungi electric conversion competence, concentrating candio-hermal and screening restrictive culture medium. Goal auxotrophy bacterial strain can be obtained quickly within 3 to 5 days by a suit system method. The deletion of large fragments avoids efficiently reversion and consanguinity recombination of exogenous plasmid. Furthermore, the method can be used many times in a same leaving strain so that multiplex auxotrophy for following genetic engineering study can be obtained because any resistance badge is not used in entire process.

The invention discloses a method for using the transcription factor Crz1p to regulate acid stress resistance of Toruula glabrata, belonging to the field of bioengineering. In the present invention, by deleting or overexpressing the Cgcrz1 gene of Torulopsis glabrata, the acid stress resistance of the strain is correspondingly reduced or improved. The present invention also compares the cell membrane fatty acid, sterol composition, ratio and permeability of the deletion mutant strain Cgcrz1Δ under acid stress. Acid stress resistance.

The present invention belongs to the field of bioengineering technology. The bacterium of the present invention is a kind of Torulopsis glabrata, WSH-, obtained with WSH-IP303 as starting strain and through NTG mutagenesis and breeding selection in culture medium with added acetic acid as replenishing carbon source. Compared with the starting strain, LQ307 has mush reduced pyruvate decarboxylase, and strong and stable pyruvate producing capacity. The yield of pyruvate reaches 46.2 g / L, 21% higher than that of starting strain when using acetic acid as replenishing carbon source and through shaking bottle culture for 48 hr; the yield of pyruvate may reach 68.7 g / L the glucose converting rate may reach 0.651 g / g through fermentation in a 5 L fermenting tank for 64 hr.

The invention discloses Weissella for promoting zygosaccharomyces rouxii to produce guaiacol substances, and belongs to the technical field of biological engineering. By co-culturing Weissella paramesenteroides LCW-28 and zygosaccharomyces rouxii ZQ-01 in a culture medium, the yield of guaiacol substances is effectively increased; the total yield of the guaiacol substances is the highest; the total yield of the guaiacol substances is increased by 3.87 times compared with the total yield of the guaiacol substances cultured independently, is increased by 15.71 times compared with candida, and is increased by 17.28 times compared with torulopsis glabrata, wherein the yield of 4-ethyl guaiacol is 2.12 mg / L, and is 8.8 times of that of the single culture of the zygosaccharomyces rouxii.

The invention discloses a method for increasing the concentration of flavor liquid fermentation components of a fen-favor liquor. The method comprises the following steps of adding a fen-favor high-temperature yeast into cooked and spread-cooled sorghum to stack; adding a liquefying enzyme and a saccharifying enzyme into a fermentation tank filled with sorghum flour paste, and adding the stacked sorghum to ferment for 2-4 days after liquefying and saccharifying; adding loofah sponge to ferment for 1.5-2.5 days; adding torulopsis glabrata to ferment for 1.5-2.5 days; connecting the fermentation tank and an overflow tower, and carrying out backflow fermentation on a fermentation mash between the fermentation tank and the overflow tower filled with carbon dioxide for 4-6 days; and stopping backflow, enabling all the fermentation mash to enter the fermentation tank, and then, adding the fen-favor high-temperature yeast to ferment for 2-4 days. A liquid-solid interface is increased by utilizing the loofah sponge, so that the formation quality of an ester substance is increased, and the flavor increasing aim is achieved; and meanwhile a gas-liquid interface is increased through the overflow of a fermentation liquid in the large overflow tower filled with carbon dioxide, and the formed flavor components are increased.

The invention discloses a method for improving acid tolerance of torulopsis glabrata, and belongs to the field of biological engineering. The final biomass of the torulopsis glabrata is increased by 59.2% under the condition of pH2.0 by Med15B overexpression, under the condition of pH2.0, the 12h cell product of the torulopsis glabrata is increased by 73.8% compared with that of original strain, the content of cryptosterol, the content of coprosterol and the content of ergosterol of cell membranes are respectively increased by 970%, 16.6% and 94.5%, the integrity and the liquidity of the cell membrane are enhanced, low pH cell tolerance is increased, growth ability is enhanced, and pyruvic acid fermentation ability is increased by 100% compared with that of the original strain.

The invention belongs to the technical field of fermentation engineering and discloses a recombinant bacterium with a function of pyruvate production intensity improvement and application of the recombinant bacterium with the function of pyruvate production intensity improvement. By targeting expression of a mitochondrion pyruvate carrier protein to a cell membrane in a torulopsis glabrata strain, cell outgoing rate of pyruvate is increased while a feedback inhibition effect of the pyruvate on glycolytic pathway key enzymes is weakened, and consequently pyruvate yield is increased, and production intensity is improved. By means of genetic engineering technology, a signal peptide sequence Shrew1p and MPC1 derived from saccharomyces cerevisiae are positioned and integrated to a pY26 expression vector to construct a saccharomyces cerevisiae recombinant expression plasmid pY26-Shrew1p-MPC1 which is electrophoretically transferred into a recipient bacterium TgU to obtain an engineering strain TgU-(pY26-Shrew1p-MPC1) high in pyruvate yield. Compared with a reference strain TgU-(pY26), the engineering strain TgU-(pY26-Shrew1p-MPC1) has the advantage that dry cell weight, glucose consumption rate, pyruvate yield and pyruvate production intensity are increased by 58.3%, 68.1%, 154.4% and 152.6% respectively.

The invention discloses a method for improving the output of pyruvic acid by a fermentation method through adding gluconic acid sodium salt, which belongs to the fermentation preparation of pyruvic acid technical field. The method adopts a strategy that during the aerobic fermentation process of Torulopsis glabrata CCTCC M202019 with glucose as a substrate, 10 to 15g / L of the gluconic acid sodiumsalt is added as a mixed carbon source when the fermentation is performed for 12 to 16h, which ensures that the output of the pyruvic acid is improved by 11.16 percent and the production period is also reduced to 44h from 48h. The research thought has universal application significance to the production of various important fermented products in industry such as organic acid, amino acid and so on.

The invention discloses a method for regulating the osmotic stress of Torulopsis glabrata, belonging to the field of bioengineering. The invention regulates the ability of resisting osmotic pressure stress of Torulopsis glabrata by overexpressing or deleting the coding gene CgRDS2 of the native transcription factor. The results showed that the deletion of CgRDS2 gene in Torulopsis glabrata, compared with the wild-type strain, could reduce the biomass by 23%, cell viability by 46%, and cell membrane integrity by 47.6% under osmotic stress conditions; after overexpressing the CgRDS2 gene , under the stress condition of 1.5M NaCl, the biomass of the strain increased by 10%, the cell activity increased by 39%, and the cell membrane integrity increased by 12.1%.

The invention relates to a pyruvic acid and L-dopa co-production process and application. The process comprises the following steps: (1) obtaining a pyruvic acid feed liquid by fermentation with torulopsis glabrata, sterilizing the feed liquid by a ceramic membrane, and removing proteins, nucleic acids, pigments and other macromolecules by an ultrafiltration membrane; (2) adding a certain amount of catechol, ammonium acetate, EDTA, sodium sulfite and other compounds to a pyruvic acid concentrate according to the molar amount of pyruvic acid, adjusting the pH to 7.0-9.0, and preparing an enzyme-catalyzed substrate solution; (3) fermenting with tyrosine-phenol lyase genetic engineering bacteria to obtain a tyrosine-phenol lyase bacterial solution, centrifuging to collect thalli, breaking cells by a high-pressure homogenizer, centrifuging, and collecting an enzyme solution; and (4) adding a certain amount of substrate solution to the enzyme solution, stirring evenly, and at the temperature of 25 DEG C, carrying out seal shock reaction. The substrate solution is prepared from the pyruvic acid concentrate, and the concentration of catechol is controlled at 0-10 g / L by fed-batching the substrate solution, the product reaches 120 g / L or more, and fed-batch of the substrate solution is stopped; when the content of catechol in the reaction solution is less than 0.2 g / L, and the reactionis stopped.

The present invention provides a genetically engineered Torulopsis glabrata with enhanced extracellular secretion of pyruvic acid. T. glabrata strain was obtained from China Center for Type Culture Collection with CCTCC No: M202019 and over-expressed the optimized CutA (SEQ ID NO:2) encoding stress protein. Both of the temperature tolerance of T. glabrata and extracellular concentration of pyruvate were improved by overexpressing the optimized CutA. The optimum growth temperature of genetically engineered T. glabrata was increased too. The present invention can be widely used to increase extracellular levels of pyruvate during the fermentation process.