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Method for synthesizing acetoin and derivative thereof through biological method

A technology of acetoin and biological method, applied in the field of molecular biology, can solve problems such as consumption and impact

Active Publication Date: 2014-04-16
QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This natural pathway uses acetolactate as a key metabolic intermediate, but acetolactate is also used in microbial metabolism to synthesize amino acids, terpenoids, etc., so it will be consumed in large quantities, which will affect acetoin and 2,3-butanedi alcohol production

Method used

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  • Method for synthesizing acetoin and derivative thereof through biological method
  • Method for synthesizing acetoin and derivative thereof through biological method
  • Method for synthesizing acetoin and derivative thereof through biological method

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Embodiment 1

[0066] The PDC gene of Zymomonas mobilis was cloned between the NdeI and BglII sites of the pET28a (purchased from Novagen) plasmid vector, and the resulting new plasmid was called pJXL65 ( Figure 4 ). Figure 4 Among them, Zm6PDC is the gene encoding pyruvate decarboxylase of Zymomonas mobilis. The yerE gene of Yersinia pseudotuberculosis was cloned between the NdeI and KpnI restriction sites of the pACYduet1 (purchased from Novagen) plasmid vector, and the resulting new plasmid was called pJXL63 ( Figure 5 ). Both pJXL65 and pJXL63 were transformed into Escherichia coli BL21(DE3) (purchased from Invitrogen) by electric shock. Culture the constructed cells in 100ml LB glucose medium (dissolve 10g sodium chloride, 10g trypsin powder and 5g yeast powder in 0.5L water for steam sterilization, dissolve 20g glucose in 0.5L water for steam sterilization, after sterilization and cooling mix the two in equal proportions). The culture temperature was maintained at 30°C. When th...

Embodiment 2

[0068] The PDC gene of Zymomonas mobilis was cloned between the NcoI and BamHI restriction sites of the pJXL63 plasmid, and the resulting new plasmid was called pJXL78 ( Image 6 ). The bdhA gene of Bacillus subtilis was cloned between the NcoI and BamHI restriction sites of the pET28a (purchased from Novagen) plasmid vector, and the resulting new plasmid was called pJXL79 ( Figure 7 ). Both pJXL78 and pJXL79 were transformed into Escherichia coli BL21(DE3) (purchased from Invitrogen) by electric shock. Culture the constructed cells in 100ml LB glucose medium (dissolve 10g sodium chloride, 10g trypsin powder and 5g yeast powder in 0.5L water for steam sterilization, dissolve 20g glucose in 0.5L water for steam sterilization, after sterilization and cooling mix the two in equal proportions). The culture temperature was maintained at 30°C. When the cells grow to a certain stage, usually the exponential growth stage, add IPTG (final concentration 50mg / L) to induce the expres...

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Abstract

The invention discloses a method for synthesizing acetoin and a derivative thereof through a biological method, which belongs to the technical field of the molecular biology. The method comprises the steps of introducing pyruvate decarboxylase genes and genes with acetoin enzyme activity into host bacteria to obtain recombination bacteria, producing the acetoin by fermenting the recombination bacteria, and further producing 2,3-butanediol on the basis of the acetoin. By adopting the method, the acetoin and the 2,3-butanediol are successfully generated by fermenting the recombined bacteria strain, and the problem that the yield of the acetoin and the 2,3-butanediol is influenced by the consumption of an intermediate product acetyl emulsion in the natural production process can be solved.

Description

technical field [0001] The invention relates to a method for synthesizing acetoin, specifically, carrying out molecular biological transformation on Escherichia coli to synthesize acetoin, and further synthesizing 2,3-butanediol on the basis of acetoin, belonging to molecular The field of biological technology. Background technique [0002] Acetoin (alias: methyl acetylmethanol, 3-hydroxy-2-butanone) has a strong creamy and fat-like aroma, and has a pleasant milky aroma after being highly diluted. It is mainly used to configure flavors and is an important food Additives and drug synthesis raw materials. Traditionally, acetoin can be obtained by reacting 2,3-butanedione with zinc under acidic conditions, or fermented from carbohydrates with fungi such as Aspergillus or Penicillium. 2,3-Butanediol is mainly used as spices and organic synthesis reagents, and it is also a potential biofuel, which can be obtained from carbohydrates fermented by Bacillus subtilis. [0003] Alth...

Claims

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Application Information

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IPC IPC(8): C12P7/26C12N15/70C12P7/18
Inventor 咸漠姜兴林刘炜徐鑫刘辉
Owner QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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