42 results about "Torulopsis glabrata" patented technology

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 construction method and use of a fumaric acid producing T.glabrata candida glabrata engineering strain and belongs to the field of fermentation engineering. In the invention, a recombinant T.glabrata FMME045 strain is obtained by overexpression of a malic dehydrogenase (RoMDH) gene and a fumarase (RoFUM1) gene from rhizopus oryzae in a uracil deficient (Ura) T.glabratadeltaura3 strain of a pyruvic acid producing strain T.glabrata in a free expression mode. When the strain is used for fermentation production of fumaric acid, the fumaric acid yield after the fermentation is performed for 60 hours is increased by 6 times to 35mg/L compared with that of a starting strain. Thus, a new approach is provided for producing fumaric acid by a microbial fermentation process. The method has a bright application prospect.

The invention discloses a high temperature-resistant yeast strain for producing pyruvic acid and a use thereof. Pyruvic acid is an important organic acid in metabolism. The high temperature-resistant yeast strain for producing pyruvic acid has wide purposes. The high temperature-resistant yeast strain is high temperature-resistant Torulopsis glabrata TIB-G90 CGMCC No. 5434 and is screened by a high-temperature adaptive evolution method. The high temperature-resistant Torulopsis glabrata TIB-G90 CGMCC No. 5434 can grow well at a temperature of 40-45 DEG C and does not influence an accumulation amount of pyruvic acid in a fermentation broth. The high temperature-resistant yeast strain can reduce a temperature reduction cost in production and has wide industrial application prospects.

The invention discloses a method for regulating acid stress resistance of torulopsis glabrata by utilizing a transcription factor Crz1p and belongs to the field of bioengineering. By deletion or overexpression of Cgcrz1 genes of torulopsis glabrata, the acid stress resistance of strains is reduced or improved correspondingly. The cell membrane fatty acid and sterol ingredients and proportion and permeability of deletion mutant strains Cgcrz1 delta under acid stress are compared, it is found that Crz1p is an essential transcription factor for torulopsis glabrata to resist the acid stress, and by the overexpression of Crz1p, the acid stress resistance of torulopsis glabrata can be improved.

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 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 invention discloses a construction method and an application technology of a Torulopsis glabrata genome-scale metabolism network model, belonging to the field of systems biology. The construction method of the model is an all-round semi-automatic construction method of combining comparative genomics annotation and bacteria-specific information of Torulopsis glabrata protein homology based on an automatic model. The invention provides a method for characterizing auxotroph characteristics in a Torulopsis glabrata metabolism network by adding necessary nutrient substances, lacked in bacteria, to a biomass equation. Alcohol dehydrogenase is knocked out by using single genes in the Torulopsis glabrata metabolism network model, and flow equilibrium analysis is applied, so that the output of pyruvic acid is increased. The construction method and the application technology of the Torulopsis glabrata genome-scale metabolism network model provide a high-efficient platform for fully understanding and reforming physiological and biochemical metabolisms of the Torulopsis glabrata.

The invention discloses Torulopsis glabrata and a method for fermenting to produce pyruvic acid by adopting a strain. The Torulopsis glabrata is an aspartic acid and lactamine defective pyruvate high yield strain, can reduce the metabolism of the pyruvic acid, and increase the conversion rate of sugar acid. In addition, in the Torulopsis glabrata, a transamination reaction from the pyruvic acid to lactamine exists, and a decarboxylic reaction from aspartic acid to the lactamine also exists. Compared with the prior art, when the strain is adopted to ferment for producing the pyruvic acid, the acid yield can be improved by 38 percent in the same production period, and the conversion rate can be improved by between 2 and 3 percent, so the strain has better prospect of industrialized production.

The invention relates to a method of boosting excessive accumulation of pyruvate by adding praline, belonging to directionally changing and optimizing microorganism metabolic function and optimizing fermentation process technical field, which is characterized in that: the torulopsis glabrata strain (T.glabrata) CCTCC M202019 of multiplex vitamin drawback is used as the starting strain, when the concentration of the sodium pyruvate is 0.4mol/L, praline is added into the culture medium and the concentration of the praline in the culture medium is 1g/L, the praline protects the torulopsis glabrata strain cells as compatible material and boosts the CCTCC M202019 to synthesize pyruvate. The method of boosting excessive accumulation of pyruvate by adding praline has the advantages that: the pyruvate fermentation cycle is shortened for 8 hours by adding 1g/L praline into the culture medium, and the pyruvate yield and production intensity is respectively enhanced for 5.4 percent and 18.6 percent; the research result can be applied to the organic acid fermentation and other typical industrial biological process, and provides new technique to industrial biotechnique, in particular the optimization of the fermentation process.

The invention discloses yeast capable of enduring high-concentration pyruvic acid and low pH and a breeding method thereof, and belongs to the field of strain breeding. In the invention, adaptive evolution technology is adopted, Torulopsis glabrata CCTCC M202019 is subjected to progressive increase of stress intensity of high-concentration sodium pyruvate by a chemostat culture system under the condition of low pH of 4.5, and an evolved strain capable of enduring high-concentration pyruvic acid and low pH stress is obtained. Under the condition that the pH is 5.5, 4.7 and 4.3 respectively, the pyruvic acid yield is 55.8g.l<-1>, 35.97g.l<-1> and 18.5g.l<-1> respectively, which is improved by 15.5 percent (48.3g.l<-1>), 51.8 percent (23.7g.l<-1>) and 83.2 percent (10.1g.l<-1>) respectively compared with that of a starting strain. The invention overcomes the sensitivity of the yeast to the low pH value and the high-concentration pyruvic acid, and has important significance for producing pyruvic acid.

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 sodium salt 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 an industrial production and fermentation technology of pyruvic acid. The technology comprises plate cultivation of torulopsis glabrata, tomato bottle cultivation, shake-flask cultivation and first-level seed tank cultivation for industrial production and fermentation technology of a fermentation tank of 150 cubic meters, and determination of specific components and ratios of a culture medium. The process blank of industrial production and fermentation of the pyruvic acid, especially fermentation and production suitable for the large fermentation tank of 150 ic metes is filled; the fermentation period is shortened to 56h; relatively high yield is also obtained; internal friction and conversion of a strain on saturated pyruvic acid of a fermentation liquid are effectively solved; and the yield and quality of a product are ensured.

The invention discloses a method for knocking out a torulopsis glabrata mitochondrion gene and belongs to the technical field of genetic engineering. In the invention, a homologous recombination fragment having the flanking sequences of an acetyl-ornithine transaminase gene (ARG 8m) and a mitochondrion gene to be knocked out is obtained by mainly using a long terminal primer, a torulopsis glabrata mitochondrion is transferred by a gene gun, and heterogeneity of the mitochondrion is eliminated by anaerobic culture to obtain a transformant which only has the gene on the modified mitochondrion. The torulopsis glabrata obtained by the invention is easy to identify and protect; the knocking method represents a novel approach and method for the genetic engineering modification of diploid of polyploidy yeasts; and the knocking method is simple and efficient, can retain the original characteristics of the yeasts, and can knock out the mitochondrion gene within 5 to 7 days.

The invention discloses a method for improving pyruvic acid yield via a fermentation process by using urea as a nitrogen source and belongs to the technical field of fermentation engineering. In the invention, multi-vitamin auxotrophic torulopsis glabrata CCTCCM202019 is used as a producing strain, urea is used in place of ammonium chloride as a unique nitrogen source, the initial added amount per liter is 7 grams, and batch fermentation culture is performed in a laboratory 7-liter fermentation tank. The method can effectively improve pyruvic acid yield (11.9 percent) and production strength (30.9 percent) and greatly reduce a fermentation period (20.6 percent); the achievements are successfully tested in a 30-liter fermentation tank; and the method has a bright industrial application prospect.