Cytochrome P450BM-3 monooxygehase varient gene and its use

Abstract

The present invention discloses cytochrome P450BM-3 monoxygenase variant gene and its use. The P450BM-3 variant gene is obtained via error-prone PCR technology mediated random mutant, and has Asp168->Asn, Ala225->Val and Lys440->Asn. The variant gene has nucleotide sequence of SEQ ID No. 1 and encods the cytochrome P450BM-3 monoxygenase. The present invention creates one kind of variant gene P450BM-3 with high catalytic activity, and the variant gene expresses mutant enzyme capable of catalyzing indole to create dye indigo blue in high activity. The present invention provides wide application foreground for the biological conversion of producing dye indigo blue.

Description

technical field [0001] The invention relates to the technical field of indigo dye production by genetically engineered bacteria in biochemical industry, in particular to a cytochrome P450BM-3 monooxygenase variant gene and its application. Background technique [0002] The directed evolution technology of enzyme molecules is to artificially create special evolutionary conditions, simulate the natural evolutionary mechanism, randomly mutate genes in vitro, and finally obtain the pre-desired enzymes with certain properties from one or more artificial mutant enzyme libraries through certain screening or selection methods Evolving enzymes with some characteristics. compared to natural evolution. The directed evolution process of enzyme molecules is completely evolved under human control, so that the enzyme molecules are directed towards the specific goals that people expect. In recent years, with the application of technologies such as error-prone PCR and DNA shuffling, under ...

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Problems solved by technology

For example, aniline and phthalic anhydride used in the synthesis of indigo can cause acute and chronic poi

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