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Yeast capable of enduring high-concentration pyruvic acid and low pH and breeding method thereof

A technology of pyruvic acid and sodium pyruvate, applied in the field of breeding high-concentration pyruvate and low pH tolerance T. glabrata, and breeding high-tolerance T. glabrata, can solve the problem of organic acid ability and speed problems such as decline, high scope of application, and equipment cost limitations, to achieve the effect of good growth performance, low professional quality requirements, and good pyruvic acid output

Inactive Publication Date: 2011-05-11
JIANGNAN UNIV
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AI Technical Summary

Problems solved by technology

[0003] Generally speaking, in the process of organic acid fermentation, in order to maintain the pH of the fermentation system in the optimum range, a certain amount of alkaline substances such as NaOH is added, and with the addition of NaOH, the osmotic pressure of the fermentation broth is continuously increased; When the osmotic pressure in the solution reaches a certain concentration, the ability and speed of the cells to further accumulate organic acids will gradually decrease, making it difficult to reach the maximum theoretical yield.
Although the combination of bioreaction and product separation technology can improve the performance and efficiency of bioreaction process, the scope of application and high equipment cost of this method limit its application in large-scale industrial production

Method used

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  • Yeast capable of enduring high-concentration pyruvic acid and low pH and breeding method thereof

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Experimental program
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Effect test

Embodiment 1

[0020] The breeding method of embodiment 1 Tosula glabrata (T.glabrata)

[0021] 1. Starting strain

[0022] The starting strain is T.glabrata CCTCC M 202019, niacin (NA), biotin (Bio), thiamine (B 1 ), pyridoxine (Pdx) and other four kinds of vitamin auxotrophy, and the activity of pyruvate decarboxylase is constitutively reduced. The strain was selected and bred for this research laboratory. CN1392246A.

[0023] 2. Breeding method

[0024] Torulopsis glabrata (Torulopsis glabrata) CCTCC M 202019 was used as the starting strain, and it was cultivated in the seed medium to the logarithmic growth phase, and placed in the evolution medium containing 20g / L sodium pyruvate and pH 4.5 for 24h Afterwards, through the chemostat culture system, according to the dilution rate of 0.1h -1 Feed the evolution medium with 20g / L sodium pyruvate, when the pH in the culture system drops to 4.0, according to the dilution rate of 0.1h -1 Feed the evolution medium containing 30g / L sodium pyr...

Embodiment 2

[0026] Example 2 Tolerus glabrata tolerant to high pyruvate concentration and low pH stress

[0027]A Torulopsis glabrata tolerant to high pyruvate concentration and low pH stress, taxonomically named Torulopsis glabrata, with a preservation number of CCTCC M 2010061, preserved in China Typical Culture on March 23, 2010 Object Collection Center.

Embodiment 3

[0028] Embodiment 3 produces pyruvic acid by fermentation

[0029] 1. Activation of strains

[0030] The starting strain T.glabrata CCTCC M 202019 and the bred strain CCTCC M2010061 were first turned to a slant and cultured in a 30°C incubator for 16 hours.

[0031] 2. Seed cultivation

[0032] The activated single colony was picked and inoculated into the seed medium, and cultured at 30° C. and 200 rpm for 24 hours.

[0033] 3. Shake flask culture

[0034] The seeds were transferred to the fermentation medium with 10% inoculation amount, and the pH was set to 5.5, 4.7, 4.3 respectively, and hydrochloric acid was used as a pH regulator, and cultured at 30°C and 200rpm for 48h. After the fermentation, the cell concentration and pyruvate production were measured, and the results are shown in Table 1.

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Abstract

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.

Description

technical field [0001] The present invention relates to a highly tolerant Torulopsis glabrata, in particular to a strain of Torulopsis glabrata that tolerates high-concentration pyruvate and low pH stresses; The method for the tolerance of Toruula glabrata, in particular to the method of breeding high-concentration pyruvic acid and low pH tolerance Toruula glabrata. Background technique [0002] The production of pyruvate by fermentation has been studied since the 1950s, and many achievements have been made. Using the principles of fermentation process optimization and metabolic engineering transformation, aiming at the unification of high yield, high yield and high production intensity of pyruvate, a large number of optimization and transformations have been carried out on the fermentation process of pyruvate produced by the multiple auxotrophic strain T.glabrata Research work has greatly increased the production of pyruvate. But with increased pyruvate production comes t...

Claims

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Application Information

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IPC IPC(8): C12N1/16C12R1/645
Inventor 陈坚汪军周景文刘立明
Owner JIANGNAN UNIV
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