Acetoin high-yield strain and application thereof in fermentation to produce acetoin

A technology of acetoin and bacilli, applied in the field of bioengineering, can solve the problems of low yield of target products, inability to be widely popularized, shortage of raw materials and the like

Active Publication Date: 2018-07-06
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional methods of chemically synthesizing acetoin mainly include the partial chlorination process of diacetyl, the selective oxidation process of 2,3-butanediol and the chlorination hydrolysis process of butanone, etc. Although these methods are easy to operate and mature in technology, the target product low yield and serious environmental pollution
The enzymatic conversion method is similar to the chemical synthesis method, and it has not fundamentally solved the shortage of raw materials, so it cannot be widely used

Method used

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  • Acetoin high-yield strain and application thereof in fermentation to produce acetoin
  • Acetoin high-yield strain and application thereof in fermentation to produce acetoin
  • Acetoin high-yield strain and application thereof in fermentation to produce acetoin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] (1) Adaptive evolution:

[0027] A. Take 200 μL of the original bacterial strain Bacillus amyloliquefaciens FMME088 and insert it into a 500mL shaker flask containing 50mL of seed medium. After cultivating at 200rpm for 12 hours at 37°C, insert it into 500mL of 50mL screening liquid medium (the concentration of acetoin is 70g / L). After culturing in a shake flask at 37°C and 200rpm for 24 hours, the bacterial solution was centrifuged at 4°C and 6000rpm for 5 minutes, and all the obtained bacteria were transferred to the screening liquid medium with acetoin concentration of 70g / L for 3 times, and then the third time was taken. After culturing for 24 hours, centrifuge at 4°C, 6000rpm for 5min, and transfer all the obtained bacteria into a 500mL shake flask containing 50mL screening liquid medium (the concentration of acetoin is 80g / L), and repeat the transfer of acetoin The screening liquid medium with a concentration of 80g / L was used for 3 times. Repeat this operation, ...

Embodiment 2

[0046] The screened bacterial strains were identified according to "Microbial Taxonomy" for their physiological and biochemical characteristics (see Tables 1 and 2 below):

[0047] Table 1 Comparison of colony morphological characteristics

[0048]

[0049] Table 2 Physiological and biochemical identification results of strains

[0050]

Embodiment 3

[0051] Example 3: Detection of acetoin tolerance of mutant strains

[0052] Step 1: Prepare medium

[0053] Seed medium in g / L: glucose 60, yeast powder 10, soybean peptone 10, beef extract 10, NaCl 0.5;

[0054] Acetoin tolerance verification medium in g / L: acetoin (0, 70, 80, 85, 90, 95, 100), glucose 20, yeast powder 12.5, peptone 12.5, KH 2 PO 4 3,K 2 HPO 4 3. MgSO 4 ·7H 2 O 0.4;

[0055] Solid medium in g / L: glucose 20, yeast powder 10, soybean peptone 10, NaCl 5, agar powder 20.

[0056] Step 2: Acetoin Tolerance Test

[0057] Take 0.2mL from the refrigerated glycerol tube and insert it into a 500mL Erlenmeyer flask containing 50mL seed medium, and culture it with shaking at 200rpm at 37°C for 12h. Inoculate seeds into 750 mL shake flasks containing 70 mL of acetoin tolerance verification medium, initial OD 600 =1.5, 37°C, 200rpm shaking culture for 6h, then add acetoin, so that the final concentration of acetoin in the medium is 0, 70, 80, 85, 90, 95 and 100g·...

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Abstract

The invention discloses an acetoin high-yield strain and application thereof in fermentation to produce acetoin and belongs to the technical field of bioengineering. Bacillus amyloliquefaciens H-5 disclosed by the invention is preserved in China Center for Type Culture Collection on 15th November 2017, a preservation address is Wuhan university in Wuhan of China, and a preservation number is CCTCCNO:M 2017694. Acetoin tolerance capacity of the acetoin high-yield strain H-5 disclosed by the invention can reach 90g/L. The H-5 is amplified and cultured on a 30L fermenting tank, culturing time is52h, an acetoin yield can reach 85.24g/L, and the acetoin yield is the maximum yield for an existing microbiological fermenting method to produce the acetoin.

Description

technical field [0001] The invention relates to a high-yield acetoin strain and its application in fermentative production of acetoin, belonging to the technical field of bioengineering. Background technique [0002] Acetoin, the chemical name is 3-hydroxy-2-butanone, also known as methyl acetylmethanol, usually the monomer is colorless or light yellow liquid, the dimer is white crystalline powder, with a special creamy aroma, natural Existing in many foods such as cocoa, cheese, bananas, grapes, corn, etc., it is a widely used food-grade spice, and it is listed as a synthetic food that is allowed to be used in GB2760-2011 (National Standards for Food Safety / Usage Standards for Food Additives). spices. At the same time, acetoin, as an important 4-carbon platform compound, is listed by the US Department of Energy as one of the 30 priority platform compounds for development and utilization. Especially chiral acetoin, as a high value-added chemical, can be widely used in the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P7/26A23L29/00A23L27/24C12R1/07
CPCA23L27/24A23L29/065C12N1/20C12P7/26C12N1/205C12R2001/07
Inventor 刘立明王诗卉罗秋玲陈修来刘佳
Owner JIANGNAN UNIV
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