Genetically engineered bacterium of Bacillus licheniformis with high yield of phenyllactic acid, method for producing phenyllactic acid and application

A technology of Bacillus licheniformis and genetically engineered bacteria, applied in the field of genes, can solve the problems of low price and profit value of target products, no patent publication documents have been found, and it is not suitable for large-scale industrial production, and achieves rich enzyme system and ability to resist extreme environments. Strong and productive effect

CN112080452AActive Publication Date: 2020-12-15TIANJIN UNIV OF SCI & TECH
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Publication Date
2020-12-15

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Abstract

The invention relates to a genetically engineered bacterium of Bacillus licheniformis with high yield of phenyllactic acid. A construction method of the genetically engineered bacterium is as follows:modifying bacillus licheniformis TCCC 11148 (2709) or / Escherichia coli Rosetta (DE3) by a genetic engineering and metabolic engineering technology. The construction method comprises three steps of molecular manipulations: inhibiting branching of chorismic acid to a tryptophan pathway; inhibiting branching of prephenic acid to a tyrosine pathway; and over-expressing phenyllactic acid synthase / aminotransferase. By modifying a Bacillus licheniformis aromatic amino acid metabolic pathway, knocking out a phenyllactic acid synthesis precursor phenylalanine synthesis pathway competitive enzyme geneand overexpressing a most direct precursor enzyme (lactic dehydrogenase) gene for phenyllactic acid synthesis, the yield of phenyllactic acid is greatly increased, and a new thought is provided for obtaining a target product phenyllactic acid. Finally, the yield of the obtained phenyllactic acid production strain reaches to 17.8 g / L after fermentation, and the strain has certain potential in industrial production of phenyllactic acid.
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Description

technical field

[0001] The invention belongs to the field of genetic technology, in particular to a genetic engineering bacterium of high-yielding bacillus licheniformis, a method for producing phenyllactic acid and an application thereof. Background technique

[0002] Phenyllactic acid (PLA), also known as 2-hydroxy-3-phenylpropionic acid, has the molecular formula C 9 h 10 o 3 . In the natural environment, phenyllactic acid is mostly produced in honey and cheese, and it is a light yellow powder after preparation, which is a small molecular substance with excellent solubility. In the prior art, the synthesis of phenyllactic acid is mainly a chemical synthesis method and a biosynthesis method, and the biosynthesis can be obtained through the metabolism of various microorganisms, the most important of which is produced by fermentation of lactic acid bacteria. Phenyllactic acid is widely used in food and drug preservatives because of its broad antibacterial spectrum, high ...

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Embodiment Construction

[0055] Below in conjunction with embodiment, the present invention is further described, and following embodiment is descriptive, not limiting, can not limit protection scope of the present invention with following embodiment.

[0056] The raw materials used in the present invention, if no special instructions, are conventional commercially available products, the methods used in the present invention, if no special instructions, are conventional methods in this area, and the quality of each material used in the present invention is routine use quality.

[0057] A high-yielding Bacillus licheniformis genetically engineered bacteria, the construction method of the genetically engineered bacteria is: transforming Bacillus licheniformis TCCC 11148 (2709) or Escherichia coli Rosetta (DE3) through genetic engineering and metabolic engineering technology, including Inhibiting the branching of chorismate to tryptophan pathway, inhibiting the branching of prephenate to tyrosine pathwa...