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Process for the production of beta-lysine

A lysine and aspartokinase technology, applied in the field of recombinant microorganisms, can solve the problems of time-consuming and expensive raw materials

Inactive Publication Date: 2009-04-01
BASF SE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the chemical synthesis of β-lysine is time-consuming, requires expensive starting materials, and produces a racemic mixture

Method used

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  • Process for the production of beta-lysine

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Experimental program
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Embodiment

[0060] 1. Cloning of the proximal Clostridium lysine 2,3-aminomutase gene

[0061] According to the conserved regions upstream and downstream of the lysine 2,3-aminomutase genes of Fusobacterium nucleatum and Thermoanaerobacter tengcongensis, a set of oligonucleotide primers were designed for In the isolation of the proximal Clostridium lysine 2,3-aminomutase gene (kamA). Using PCR primers WKJ90 / WKJ65 and WKJ68 / WKJ93, and using the chromosome of Clostridium axeum as a template, the DNA fragments of the upstream and downstream regions, including the N-terminal and C-terminal sequences of the kamA gene, were amplified, respectively. After the sequence analysis of the amplified DNA fragment is completed, it is purified to obtain a product containing the start and end sequences of the kamA structural region. Based on the determined upstream and downstream sequences, PCR primers WKJ105 / WKJ106 were synthesized for isolating the full sequence of the Clostridium proximal kamA gene. ...

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Abstract

Process for the production of -lysine by constructing a recombinant microorganism which has a deregulated lysine 2,3-aminomutase gene and at least one deregulated gene selected from the group (i) which consists of aspartokinase, aspartatesemialdehyde dehydrogenase, dihydrodipicolinate synthase, dihydrodipicolinate reductase, tetrahydrodipicolinate succinylase, succinyl-amino-ketopimelate transaminase, succinyl-diamino-pimelate desuccinylase, diaminopimelate epimerase, diaminopimelate dehydrogenase, arginyl-tRNA synthetase, diaminopimelate decarboxylase, pyruvate carboxylase, phosphoenolpyruvate carboxylase, glucose-6-phosphate dehydrogenase, transketolase, transaldolase, 6-phosphogluconolactonase, fructose 1,6-biphosphatase, homoserine dehydrogenase, phophoenolpyruvate carboxykinase, succinyl-CoA synthetase, methylmalonyl-CoA mutase, provided that if aspartokinase is deregulated as gene (i) at least a second gene (i) other than aspartokinase has to be deregulated, and cultivating said microorganism.

Description

technical field [0001] The present invention relates to a method for producing β-lysine. More specifically, the present invention relates to the use of recombinant microorganisms comprising deregulated forms of DNA molecules necessary for the production of β-lysine. Background technique [0002] Although β-amino acids are rare compared to the corresponding α-amino acids, they still occur in nature both in free form and in polypeptides. Cardillo and Tomasini, Chem. Soc. Rev. 25:77 (1996); Sewald, Amino Acids 11:397 (1996). Since β-amino acids are stronger bases and weaker acids than the corresponding α-amino acids, polypeptides containing β-amino acids instead of α-amino acids have different backbone atom patterns, leading to new properties. [0003] In the 1950s, L-β-lysine was identified in several strongly basic peptide antibiotics produced by Streptomyces. Antibiotics that produce L-β-lysine by hydrolysis include puromycin, aflatoxin A, violacein, rosethricin, and dext...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/08C12P13/00C12N15/63C12N15/77
CPCC12P13/08C12P13/02C12P13/005C12N15/63
Inventor O·策尔德尔W·K·曾C·克洛普罗格A·赫罗尔德H·施罗德
Owner BASF SE
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