Biosynthesis method of levodopa

Abstract

The invention discloses a biosynthesis method of levodopa, and belongs to the technical field of biological engineering. The method constructs a recombinant expression vector pJJ-dhtpl containing a tyrosine phenol lyase gene dhptl, and realizes the high-efficiency expression of the tyrosine phenol lyase in Escherichia coli BL21. Recombinant Escherichia coli resting cells expressing tyrosine phenollyase can be used to catalyze synthesis of levodopa from cheap catechol and ammonium pyruvate in one step under the conditions of 16 DEG C, pH 7.5-8.0, and vacuum. Levodopa is difficult to dissolve in water, and can be precipitated in the form of crystals in the reaction system, the reaction is carried out thoroughly, and a molar conversion rate of the major reactant, catechol, is up to 96% or more. The recombinant Escherichia coli resting cells expressing tyrosine phenol lyase have good catalytic stability, and levodopa can be continuously accumulated by adding catechol and ammonium pyruvatemultiple times and in batches. The production process is simple, the raw materials are easy to obtain, and the product is easy to separate and purify, so that the method has a good application prospect for large-scale industrial production.

Description

technical field [0001] The invention belongs to the technical field of bioengineering and relates to a biosynthesis method of levodopa. Background technique [0002] Levodopa (levodopa, L-DOPA, 3-hydroxy-L-tyrosine), is an important compound used in the treatment of Parkinson's disease. At present, sources of levodopa include plant extraction, chemical synthesis, and biosynthesis. (1) Plant extraction method. Levodopa is found in many natural plants. In 1913, biochemist Guggenheim isolated and extracted levodopa from the capsule of broad bean for the first time. After that, the existence of levodopa was found in many plants, such as cat beans, quinoa beans, etc. Raw materials, the mass fraction of levodopa is as high as 6% to 9%. BeheraAnindita et al. (Inventi Rapid PharmAnalysis&Quality Assurance, 2011 (2): 135-146) through the improvement of extraction technology, the extraction yield of levodopa in cat beans has increased from 1.5% to 3.4%, and the purity has reached...

AI Technical Summary

Problems solved by technology

[0003] Biological synthesis of levodopa is green and safe, and is favored by many researchers, but so far most of it is limited to laboratory research, with low catalytic efficiency and limited output

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