Method for restructuring/breeding Actinobacillus succinogenes strain, and method for producing succinic acid by fermenting Actinobacillus succinogenes strain

Abstract

The invention relates to an Actinobacillus succinogenes strain CGMCC 2653 (namely F3-10) with better sodium-ion tolerance and acid-producing performance, a method for breeding the strain and a method for producing succinic acid by fermenting the strain. Actinobacillus succinogenes CGMCC 1593 is taken as an original strain and then subjected to X-ray mutation, ultraviolet mutation, diethyl sulfate(EMS) chemical mutation and nitrosoguanidine (NTG) chemical mutation respectively; three strains X-8, UV-17 and SE-6 with improved fermentative acid-producing performance and particularly improved sodium-ion tolerance, as well as a high-yield strain SF-9 with fluoroacetate resistance, are obtained by screening the obtained product; and a genome restructuring method is used for breeding the strains so as to obtain the strain F3-10 with high yield, sodium tolerance and fluoroacetate resistance. The strain F3-10 takes sugarcane molasses as raw material, adopts Na2CO3 to control fermentation pH, and performs fed-batch fermentation in a 5L fermenter, and produces 53.96 grams of succinic acid per liter in 48 hours; the yield of consumed sugar is 89.2 percent; the utilization rate of sugar is 94.0 percent; and the ratio of succinic acid to heteroacid is 6.58:1. Therefore, the strain F3-10 is remarkably improved as compared with the original strain CGMCC 1593.

Description

Actinobacillus succinicum strain reorganization, breeding method and method for producing succinic acid by fermentation technical field The invention belongs to the technical field of bioengineering, and specifically relates to a strain of Actinobacillus succinogenes CGMCC2653 (namely F3-10) with strong sodium ion tolerance and acid production performance, the breeding method of the strain and the application of the strain in anaerobic A method for the production of succinic acid by oxygen fermentation. Background technique Succinic acid, also known as succinic acid, has a molecular formula of C4H6O4 and a molecular weight of 118.09. It is a common natural organic acid and widely exists in the human body, animals, plants and microorganisms. Succinic acid is an important four-carbon compound in industry. It is widely used in food, medicine, pesticides, dyes, spices, paints, plastics and material industries as organic synthesis raw materials, intermediate products, and profe...

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Problems solved by technology

The environmental tolerance of microorganisms, such as the genetic coding mechanism of phenotypes such as acid resistance and heat resistance, is not yet fully understood. It is difficult to modify the traits controlled by multiple genes scattered on the genome by direct genetic engineering technology; However, the forward mutation rate of traditional mutagenesis is very low, and the amount of screening required to improve a strain is large and the screening time is long

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