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Method of synthesizing chirality biology by recycling viable cell as biological catalyst

A biocatalyst and biosynthesis technology, applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve problems such as difficult to achieve simultaneously, and no extraction of polar products biotransformation is found.

Inactive Publication Date: 2011-03-23
SHANGHAI JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] After searching the prior art documents, it was found that Zhilong Wang, "The potential of cloudpoint system as a novel two-phase partitioning system for biotransformation.Applied microbiology and biotechnology." 2007, 75: 1-10. (Wang Zhilong. "Turbidity System Potential for biotransformation as a new two-phase distribution system". "Applied Microbiology and Biotechnology", 2007, 75: 1-10) introduced that in the extraction biotransformation process of the water-organic solvent two-phase distribution system Polar products and maintenance of cell activity, but often difficult to achieve simultaneously
In further search, there is no literature report about the biotransformation of extracted polar products in the turbidity system of the present invention

Method used

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  • Method of synthesizing chirality biology by recycling viable cell as biological catalyst
  • Method of synthesizing chirality biology by recycling viable cell as biological catalyst

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] The ratio of Triton X-45 and Triton X-114 is 1: The mixed surfactant solution and the culture medium are 1:10 by volume to form a cloud point system with a volume of 22ml. The culture medium is composed of 2g peptone, 2g yeast powder, 2g glucose or 9g glucose, 1g yeast powder, and 100ml water. 1g(NH 4 ) 2 SO 4 , 0.3g KH 2 PO 4 , 0.2g Na 2 HPO 4 , 0.1g MgSO 4 and 0.005g CaCl 2 . Put the cloud point system into a 50ml shake flask and inoculate yeast cells at 30°C and culture in a shaker at 200rpm for 2 days. After centrifugation, the wet cell weight was determined to be 1.7g. This indicates that the cloud point system is biocompatible.

Embodiment 2

[0045] The medium consists of 2g peptone, 2g yeast powder, 2g glucose or 9g glucose, 1g yeast powder, 1g (NH 4 ) 2 SO 4 , 0.3g KH 2 PO 4 , 0.2g Na 2 HPO 4 , 0.1g MgSO 4 and 0.005gCaCl 2 . Add 8 g of PEG 20000 and 7.5 ml of surfactant Triton X-100 to 100 ml of medium to form a cloud point system. Put 20ml of the cloud point system into a 50ml shake flask and inoculate yeast cells at 30°C in a shaker at 200rpm for 2 days. After centrifugation, the wet cell weight was determined to be 1.5g. This indicates that the cloud point system is biocompatible.

Embodiment 3

[0047] In the cloud point system in Example 1, 22ml of the cloud point system was packed into a 50ml shake flask to inoculate yeast cells, and the substrate acetophenone 1% (v / v) concentration was added, at 30°C, shaking at 200rpm They were cultured in the bed for 2 days, and the wet cell weight was determined to be 1.2 g after centrifugation. This indicates that the cloud point system can tolerate high concentrations of toxic substrates.

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PUM

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Abstract

A chiral biosynthetic method, which repeatedly uses living cells as the biocatalyst, belongs to the biological engineering technical field. The procedures include that; a cloud point system is constructed, consisting of nonionic surfactant and a microorganism substrate, or of polymer, nonionic surfactant and a microorganism substrate, and the obtained cloud point system is put into a shake flask and inoculated with fresh seeds of breast yeast to yield the production, which is cultivated in a rocking bed, the cells are harvested and centrifuged with the wet weight being measured, which is usedto analyze the biocompatibility within the variation of concentration ratio of substrate / product; then the microorganism substrate inoculated with the fresh seeds of bread yeast is put into a shake flask and cultivated in a rocking bed, the cells are harvested and centrifuged with the wet weight being measured, and then the harvested fresh cells are added into the cloud point system with the biocompatibility as proved by step two, to undergo the reaction with the added substrate. The invention realizes the repeated utilization of living cells as the biocatalyst in high concentration ratio of substrate / product.

Description

technical field [0001] The invention relates to a synthesis method in the technical field of bioengineering, in particular to a chiral biosynthesis method for reusing living cells as biocatalysts. Background technique [0002] The stereoselectivity of biocatalysts enables the biosynthesis of chiral compounds with potential industrial applications in medicine, food, agrochemistry, and cosmetics. The use of living cells as biocatalysts can not only avoid the cumbersome steps of preparing purified enzymes, but more importantly, the use of living cells as biocatalysts is beneficial to multi-enzyme catalyzed or biotransformation processes that require coenzymes. [0003] Baker's yeast has potential industrial value as a biocatalyst because of its fast growth, simple culture conditions, non-pathogenic microorganisms, and multiple enzyme catalytic systems. The use of living cells to metabolize glucose to regenerate coenzymes to convert aromatic ketones to synthesize chiral aromati...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P7/02C12R1/85C12P1/00
CPCY02P20/582
Inventor 王志龙张文智王丽齐瀚实
Owner SHANGHAI JIAOTONG UNIV