Bioconversion method for anti-AIDS-medicine-reyataz midbody

Abstract

The invention discloses a bioconversion method for anti-AIDS-medicine-reyataz midbody, and belongs to the technical field of medical biology. Secondary structures of genes and codon usage biases are adjusted, the length of primers is controlled within 60 base, the primers are obtained, the obtained primers are dissolved with double distilled water, then the dissolved primers are added into a reaction system, a prepared PCR reaction system is amplified, obtained DNA fragments are subjected to gel cutting purification, and then are cloned into NdeI / XhoI loci of pET30a with a homologous recombination method, a successfully-sequenced DNA sequence is SEQ ID NO.3, and is named as Sst-2, and the corresponding amino acid sequence of the Sst-2 is SEQ ID NO.4. The bioconversion method is mild in reaction condition, and has the low requirement for equipment, the high temperature or cooling is not required in the production process, energy consumption is low, purification is convenient, single enzyme catalysis is used in the whole system, the cycle number of coenzyme is high, and the reaction condition is mild.

Description

technical field [0001] The invention relates to a preparation method of an atazanavir intermediate, in particular to a biotransformation method of an anti-AIDS drug atazanavir intermediate, belonging to the field of medical biotechnology. Background technique [0002] Atazanavir, sold under the trade name Reyataz, is an antiretroviral drug used for the treatment and prevention of HIV / AIDS. It is currently one of the main anti-AIDS drugs in the world. On the list, it is the most important drug needed by the basic health system. Bristol-Myers Squibb first developed atazanavir, which was publicly introduced in Chinese patent CN10282508C. The US FDA approved it for marketing in 2003, and China also in 2007 Approved for its listing. [0003] (2R,3S)-1-Chloro-3-tert-butoxycarbonylamino-4-phenyl-2-butanol is used as a key intermediate for the preparation of atazanavir. At present, the main production processes include chemical synthesis and biosynthesis There are two methods, and...

AI Technical Summary

Problems solved by technology

[0007] In the process of international patent WO2011005527, due to the low enzyme activity of the enzyme used, 0.3g NAD is needed to catalyze 90 grams of substrate, and the ratio of substrate: NAD is only 300, resulting in an excessive proportion of NAD cost per kilogram of product produced , so that the production cost is high. In the Chinese patent CN102732579, Saccharomyces cerevisiae is used for whole-cell catalysis, but too many yeast cells are required (10g dry cells), and the catalytic substrate can only be completely converted when it is not greater than 0.1mM. This process production efficiency is low

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