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Pyranose oxidase gene and preparation, pichia pastoris constructed from the same and application

A Pichia pastoris and oxidase technology, applied in Pichia pastoris and application fields, can solve the problems of low expression and the like

Active Publication Date: 2019-02-01
河北省微生物研究所有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] After reviewing the literature, the applicant found that the cloning and expression of the pyranose oxidase gene has only been carried out in Escherichia coli and has been successfully realized so far. For example, Zhang Keke of Hunan University and others have used E. Although the activity can be detected, the expression level is extremely low

Method used

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  • Pyranose oxidase gene and preparation, pichia pastoris constructed from the same and application
  • Pyranose oxidase gene and preparation, pichia pastoris constructed from the same and application
  • Pyranose oxidase gene and preparation, pichia pastoris constructed from the same and application

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Embodiment

[0042] 1. Pyranose oxidase gene, its gene sequence is:

[0043]GAATTCATGTCTACTTCCTCCTCCGATCCATTCTTCAACTTTACCAAGTCCTCCTTCAGATCCGCT GCTGCTCAAAAGGCTTCTGCTACTTCTTTGCCACCATTGCCAGGTCCAGACAAGAAAGTTCCAGGT ATGGACATCAAGTACGACGTGGTTATCGTTGGTTCCGGTCCAATCGGTTGTACTTACGCTAGAGAA TTGGTTGAGGCCGGTTACAAGGTTGCCATGTTCGACATTGGTGAGATCGACTCCGGTTTGAAGATT GGTGCTCACAAGAAGAACACCGTCGAGTACCAGAAGAACATCGACAAGTTCGTCAACGTCATCCAG GGTCAGTTGATGTCTGTTTCCGTTCCAGTTAACACCCTGGTCATCGATACTTTGTCTCCAACTTCT TGGCAGGCCTCCTCATTCTTCGTTAGGAACGGTTCTAACCCAGAGCAGGACCCATTGAGAAACTTG TCTGGTCAGGCTGTTACCAGAGTTGTTGGTGGTATGTCTACTCACTGGACTTGTGCTACTCCAAGA TTCGACAGAGAGCAAAGACCATTGCTGGTTAAGGATGACACTGATGCTGATGACGCTGAATGGGAC AGACTGTACACTAAGGCTGAGTCCTACTTCAAGACTGGTACTGACCAGTTCAAAGAGTCCATCAGA CACAACCTGGTCTTGAACAAGTTGGCCGAAGAGTACAAGGGTCAGAGAGACTTCCAACAGATTCCA TTGGCTGCCACTAGAAGATCCCCAACTTTTGTTGAATGGTCCTCCGCCAACACCGTTTTCGACTTG CAAAACAGACCAAACACTGACGCTCCAAACGAGAGATTCAACTTGTTTCCAGCTGTTGCCTGTGAG AGAGTCGTTAGAAACACTTCCAACTCCGAGATCGAGTCCTTGCACATTCACGACTTGATT...

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Abstract

Belonging to preparation of new genes and construction of engineering bacteria, the invention in particular relates to a pyranose oxidase gene, pichia pastoris constructed from the same and application. According to the invention, codon optimization software is mainly utilized to optimize the codons of pyranose oxidase to make them all become pichia pastoris preferred codons, and then the means ofgene synthesis is utilized to synthesize the target gene; and the gene is transformed into pichia pastoris to achieve high efficiency expression of the target protein. The recombinant pichia pastorisprepared according to the invention has high fermentation enzyme activity, lays a good foundation for application of pyranose oxidase, effectively avoids the disadvantages generated during expressionin escherichia coli, and opens up a brand new industrial efficient production way for pyranose oxidase, etc.

Description

technical field [0001] The invention belongs to the preparation of new genes and the construction of engineering bacteria, in particular to a pyranose oxidase gene and its preparation, Pichia pastoris constructed by using it and its application. Background technique [0002] Pyranose oxidase (pyranose / oxygen 2-oxidase, P2O; EC 1.1.3.10) belongs to the glucose-methanol-choline (glucose-methanol-choline, GMC) oxidoreductase family, is a prosthetic group of FAD Flavoproteinase. It can catalyze the oxidation of the C-2 position of several pyranose sugars such as glucose to generate the corresponding 2-ketose sugar, and convert O 2 reduced to H 2 o 2 . [0003] Since P2O can catalyze various sugars to generate corresponding ketoses, P2O has important application value in sugar chemistry. P2O conversion of some common sugars can provide a large number of intermediates for the production of a variety of rare sugars, fine chemicals and pharmaceuticals. P2O can also be applied ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/53C12N9/04C12N15/81C12N1/19C12N15/66C12R1/84
CPCC12N9/0006C12N15/66C12N15/815C12Y101/0301
Inventor 王玥高庆华刘蕾董聪王庆庆罗同阳马清河
Owner 河北省微生物研究所有限公司