P450BM-3 variant gene for catalyzing indigo blue generated by benzazole and and use thereof

Abstract

This invention relates to a variant gene P450BM-3 to catalytic generation of indigo blue from indole and its use. Due to random mutation caused by PCR process, variant gene P450NM-3 is obtained by screening. ATC of Ile on its 39th digit is mutated into GTC of Val. Its gene sequence is listed in SEQ.ID.No.1 of nucleotide sequence. Its variant protease can catalyze indole to generate indigo blue. Compared with a variant protease with three mutation points, which is obtained by R.D.Schmid' team, its activation increases 1.2-1.4 times in an example.

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 P450BM-3 variant gene capable of catalyzing indole to generate indigo and its use. 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 a pre-expected certain enzyme from one or more artificial mutant enzyme libraries through certain screening or selection methods. Evolutionary enzymes with some properties. compared to natural evolution. The directed evolution process of enzyme molecules is completely under human control, making the enzyme molecules move towards the specific goals that people expect. In recent years, with the application of technologies such as error-prone PCR and DNA shuffling, under the condition of eff...

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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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