A kind of recombinant Escherichia coli, preparation method and method for synthesizing d-1,2,4-butanetriol

Abstract

The invention relates to recombinant Escherichia coli, a preparation method thereof and a method to synthesize D-1,2,4-butantriol via the recombinant Escherichia coli, and belongs to the technical field of biology. XylA, yjhH and yage are knocked out of Escherichia coli; more than one of 2-ketonic acid reductase gene, methylglyoxal synthase gene, transaminase gene, transhydrogenase gene, acetokinase gene and phosphoric acid transacetylase gene is further knocked out; and / or more than one of xylonic anhydrase gene, alcohol dehydrogenase gene and glucose 6-phosphate dehydrogenase gene are over-expressed to obtain recombinant Escherichia coli. Xylose is used as a basic substrate, and glucose, acetic acid and glycerol can also be added to form a composite substrate; the synthesis of D-1,2,4- the recombinant Escherichia coli butanethiol has every low content of auxiliary products; the preparation method is simple, the production cycle is short, the cost is low, and the preparation method has a promising industrial development and application prospect.

Description

technical field [0001] The invention relates to a recombinant Escherichia coli, a preparation method and a method for synthesizing D-1,2,4-butanetriol, belonging to the field of biotechnology. Background technique [0002] D-1,2,4-Butanetriol is an important fine chemical product, which is widely used in military industry, medicine, tobacco, cosmetics, papermaking, agriculture and polymer materials and other fields. In the field of military industry, the nitro compound 1,2,4-butanetriol trinitrate of D-1,2,4-butanetriol is a good energetic plasticizer, which can replace nitroglycerin as a nitrate extender. Plastic polyether propellant (NEPE) propellant and high-energy, high-tech formula propellant. The impact, toxicity, volatility and hygroscopicity of 1,2,4-butanetriol trinitrate are lower than those of nitroglycerin, and it has better thermal stability, so it can be mixed with other energetic plasticizers. There are asymmetric carbon atoms in the molecular structure of 1...

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Problems solved by technology

[0007] (2) Pay too much attention to the biosynthesis pathway of butanetriol, ignoring the growth of host bacteria

[0008] (3) The effect of intracellular reduced coenzyme I (NADH) / reduced coenzyme II levels on the synthesis of butanetriol was not considered

[0009] In 2015, Gao Haijun from Beijing Institute of Technology (Ma Pengfei, Meng Jian, Zhou Jing, Gao Haijun. Recombinant Escherichia coli uses D-xylose to synthesize D-1,2,4-butanetriol. Acta Chemical Industry Vol.66 No. 7.2015.7: 2620-2627.) By cloning and expressing Pseudomonas putida ATCC12633 2-keto acid decarboxylase gene mdlC and Crescentus crescentus (Caulobacter crescentus CB15) D-xylose dehydrogenase gene xdh, Knockout of xylose isomerase gene xylA, 2-ketoacid aldolase gene yjhH and 2-ketoacid aldolase gene in xylose utilization and D-1,2,4-butanetriol synthesis intermediate metabolite decomposition pathway The enzyme gene yagE reconstructed the metabolic network of Escherichia coli and obtained the Escherichia coli strain MJ133K-1 that can convert D-xylose into D-1,2,4-butanetriol. On this basis, the relationship between glucose utilization and Relationship of butanetriol synthesis by knockdown of glucose-specific transporter II CB Glc The phosphoenolpyruvate (phosphoenolpyruvate: sugar phosphotransferase system, PTS) system of the recombinant strain was transformed with the gene ptsG, and the obtained recombinant E. coli strain MJ134K-1 can transform xylose while utilizing glucose for growth, and has more High synthesis ability; but still need to further improve the synthesis amount and conversion rate of butanetriol

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