Three-enzyme co-expression recombinant bacterium and application thereof in synthesis of (S)-citronellol
A technology of recombinant bacteria and co-expression, applied in applications, recombinant DNA technology, enzymes, etc., to achieve the effect of simple operation, high selectivity, and mild conditions
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Embodiment 1
[0035] Embodiment 1, acquisition of old yellow enzyme NemR-PS coding gene
[0036] Utilize the published source from Providencia stuartii (Providencia stuartii; Department of Infectious Diseases, National Institute of Health Dr. Ricardo Jorge, Lisbon, Portugal) old yellow enzyme NemR-PS coding gene (GenBank accession number is KNZ86848), After codon optimization, artificially synthesized (Hangzhou Qingke Biotechnology Co., Ltd. provides gene synthesis service) the gene encoding the old yellow enzyme NemR-PS, the nucleotide sequence and amino acid sequence are respectively as SEQ ID NO.1 and SEQ ID NO. 2.
Embodiment 2
[0037] Example 2, Acquisition of Alcohol Dehydrogenase Encoding Gene
[0038] Utilizing the published gene encoding alcohol dehydrogenase YsADH (GenBank accession number KF887947) derived from Yorkella sp. CCTCC NO.M2013099 (disclosed in patent application 2013101888839), after codon optimization, the Synthesis (Suzhou Jinweizhi Biotechnology Co., Ltd. provides gene synthesis service) the gene encoding alcohol dehydrogenase, the nucleotide sequence and amino acid sequence are respectively shown in SEQ ID NO.3 and SEQ ID NO.4.
Embodiment 3
[0039] Embodiment 3, the acquisition of the gene encoding glucose dehydrogenase
[0040] The GenBank accession number of glucose dehydrogenase BmGDH derived from Bacillus megaterium is AAA22475, and the amino acid sequence of glucose dehydrogenase BmGDH was replaced by Q252L / E170K / S100P / K166R / V72I / K137R to obtain mutant BmGDH M6 (Already disclosed in patent application 2020103075429). mutant BmGDH M6 The coding gene was artificially synthesized after codon optimization (Hangzhou Qingke Biotechnology Co., Ltd. provides gene synthesis service), and the nucleotide sequence and amino acid sequence are respectively shown in SEQ ID NO.5 and SEQ ID NO.6.
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