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Torulopsis glabrata mutant strain and application thereof in fermentation and production of pyruvic acid

A technology of T. glabrata and mutant strains, applied in fermentation, microorganism-based methods, microorganisms, etc., can solve problems such as no problems, and achieve the effects of reducing pollution, high yield and strong operability

Inactive Publication Date: 2010-03-03
SHENZHOUSPACEBIOTECHGRP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there are many patents related to pyruvate fermentation. Although its yield and conversion rate are high, there is no verification of the target product after extraction.

Method used

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  • Torulopsis glabrata mutant strain and application thereof in fermentation and production of pyruvic acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Screening process of mutant Torulopsis glabrata CGMCC No.3077

[0032] At 25°C, the starting strain Torulopsis glabrata (Torulopsis glabrata) NBRC 0005 was added to YM liquid medium (glucose mass percent 1.0%, peptone mass percent 0.5%, yeast extract mass percent 0.3%, malt extract mass percent 0.3%) ) to the mid-logarithmic phase, aseptically collected the cells, washed twice with saline, and counted with a hemocytometer, adjusting the cell concentration to 10 7 pieces / ml. Take 10ml of bacterial suspension and place it in a φ9cm plate (with magnetic stirring), and place it under a 15W UV lamp (before the experiment, turn on the UV lamp for 20 minutes to stabilize the UV light), and the vertical distance between the lamp and the bacterial solution is 30cm, turn on the magnetic stirring, irradiate for 60, 120, 180, 240, 300s respectively, take samples for serial dilution, use the pouring method, and count the living cells on the plate. The calculation formul...

Embodiment 2

[0033] Example 2: Comparison of pyruvate production by shake flask fermentation of Torulopsis glabrata NBRC 0005 and its mutant strain CGMCC No.3077.

[0034] Torulopsis glabrata (Torulopsis glabrata) NBRC 0005 and its mutant strain CGMCCNo.3077 activated by liquid medium YEPD (glucose mass percentage 2.0%, peptone mass percentage 2.0%, yeast extract mass percentage 1.0%) at an inoculum size of 10% Inoculated in the following fermentation liquid medium respectively: 10% by mass of glucose, 1% by mass of fish peptone, 0.01% by mass of magnesium sulfate, 1% by mass of calcium carbonate, 0.0000001% by mass of niacin, and 0.001% by mass of biotin , pH natural. Fill 50ml of medium into a 250ml Erlenmeyer flask. Cultivate and ferment on a shaker at 29° C. and 200 rpm for 3 days, and the product is calcium pyruvate. After measuring the content of pyruvate in the supernatant of the fermentation broth after centrifugation, the yields of the starting strain and the mutant strain were ...

Embodiment 3

[0036] Example 3: Comparison of pyruvate production by shake flask fermentation of Torulopsis glabrata NBRC 0005 and its mutant strain CGMCC No.3077.

[0037] Torulopsis glabrata (Torulopsis glabrata) NBRC 0005 and its mutant strain CGMCCNo.3077 activated by liquid medium YEPD (glucose mass percentage 2.0%, peptone mass percentage 2.0%, yeast extract mass percentage 1.0%) at an inoculum size of 10% Inoculated in the following fermentation liquid medium respectively: 15% by mass of sucrose, 2% by mass of soybean peptone, 0.05% by mass of dipotassium hydrogen phosphate, 2% by mass of calcium carbonate, 0.00001% by mass of vitamin B1, and 0.00001% by mass of vitamin B6 0.001%, pH natural. Fill 50ml of medium into a 250ml Erlenmeyer flask. Cultivate and ferment on a shaker at 29° C. and 200 rpm for 3 days, and the product is calcium pyruvate. After measuring the content of pyruvate in the supernatant of the fermentation broth after centrifugation, the yields of the starting stra...

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Abstract

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.

Description

technical field [0001] The invention belongs to the field of fermentation engineering, in particular to a method for fermenting and producing pyruvate by Torulopsis glabrata CGMCC No. 3077. Background technique [0002] Pyruvate is a very versatile organic acid that can be used as a precursor for the synthesis of many useful compounds. In the pharmaceutical industry, pyruvate is used as L-tryptophan, L-tyrosine, L-dopa, L-cysteine, L-leucine, vitamin B 6 and B 12 It is the main raw material in the enzymatic synthesis process; in terms of pesticides, it is used as the starting material of various pesticides such as atropic acid and grain protectants; ) and cosmetic antioxidants. Pyruvate is also widely used as biotechnology diagnostic reagents and detection reagents, which can be used for the determination of primary alcohols and secondary alcohols, the determination of transaminases in biochemical research, and the color reagent of aliphatic amines. In addition, pyruvate...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12P7/40C12R1/88
Inventor 王淑豪石萌萌王嘉沈强庞欣
Owner SHENZHOUSPACEBIOTECHGRP
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