Bacillus subtilis for highly producing D-ribose and co-producing acetoin

A technology of Bacillus subtilis and acetoin, which is applied in the directions of bacteria, microorganism-based methods, biochemical equipment and methods, etc., can solve the problems of low utilization rate of raw materials, high cost, restrictions on raw material sources and prices, etc., and achieves cultivation conditions. The effect of extensive, high mutation rate and stable product yield

Inactive Publication Date: 2011-02-16
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

No matter chemical synthesis method or enzymatic conversion method to produce acetoin, diacetyl or 2,3-butanediol, etc. are used as raw materials. These two substances are also synthetic fragrances rather than bulk chemical products. The sources and prices of raw materials are different It is limited, which may be one of the reasons why 3-hydroxybutanone has not been well researched and developed at present. Therefore, the active development of 3-hydroxybutanone by microbial fermentation using sugar as raw material is expected to change this situation
[0005] To sum up, the existing technologies all use Bacillus subtilis culture to produce D-ribose alone, and the acetoin involved is used as a by-product for metabolic regulation theory research, or acetoin is produced alone, There are problems such as low utilization rate of raw materials and high cost

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Incline medium: 5g / L D-sorbitol, 10g / L peptone, 2g / L yeast extract, 2g / L NaCl, 20g / L agar, pH 6.9.

[0043] Seed medium: 20g / L glucose, 20g / L corn steep liquor, 3g / LK 2 HPO 4 , 1g / LKH 2 PO 4 , 1g / L yeast extract, pH 6.9.

[0044] Fermentation medium: 200g / L glucose, 8g / L corn steep liquor, 4g / L yeast powder, 7g / L (NH 4 ) 2 SO 4 , 0.05g / LMnSO 4 , 3g / LCaCO 3 , 1.5g / L sorbic acid, pH 6.9; 3L of fermentation broth in a 5L fermenter, sterilized at 121°C for 15 minutes.

[0045] Put two loops of Bacillus subtilis CGMCC No.3720 in the seed medium, cultivate at 36°C for 18 hours to obtain a mature seed liquid, and insert the seed liquid into the cooling medium according to the inoculation amount of 10% (v / v) of the fermentation liquid volume In the final fermentation broth, dissolved oxygen was controlled at 1.1vvm, 600rpm, and fermented at 37°C for 72 hours. The content of D-ribose and acetoin in the finally obtained fermentation broth were 113g / L and 30.5g / L respecti...

Embodiment 2

[0051] Fermentation medium: 150g / L glucose, 50g / L xylose, 5g / L corn steep liquor, 6g / L yeast extract, 5g / L (NH 4 ) 2 SO 4 , 0.05g / L MnSO 4 , 2g / LCaCO 3 , 0.4g / L sorbic acid, pH 6.9; 3L of fermentation broth in a 5L fermenter, sterilized at 121°C for 15 minutes.

[0052] Prepare seed culture medium, formula and culture method are with embodiment 1, seed liquid is inserted in the fermentation culture liquid after cooling by the inoculum size of 10% (v / v) of fermented liquid volume, dissolved oxygen control is 1vvm, 500rpm, Under 37 ℃ of fermentation 72h, the D-ribose content in the fermented liquid obtained at last is 98.51g / L, and the acetoin content is 29.1g / L, respectively compared with only using common culture medium (that is, does not add sorbic acid, but other components The same) yield increased by 12.95% and 29.33%.

Embodiment 3

[0054] Fermentation medium: 20g / L glucose, 20g / L molasses, 1g / L corn steep liquor, 0.5g / L peptone, 0.5g / L yeast powder, 0.5g / L (NH 4 ) 2 SO 4 , 0.02g / L MnSO 4 , 0.01g / LMgSO 4 , 0.01g / LCaCO 3 , 3.5g / L sorbic acid, pH 6.9; 3L of fermentation broth in a 5L fermenter, sterilized at 121°C for 15 minutes.

[0055] Prepare seed culture medium, formula and culture method are with embodiment 1, seed liquid is inserted in the fermentation culture liquid after cooling by the inoculum size of 10% (v / v) of fermented liquid volume, dissolved oxygen control is 1vvm, 550rpm, Under 37 ℃ of fermentation 70h, the D-ribose content in the fermented liquid obtained at last is 91.4g / L, and the acetoin content is 25.5g / L, which are respectively compared with only using common culture medium (that is, without adding sorbic acid, but other components The same) yield increased by 4.80% and 13.33%.

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Abstract

The invention discloses a bacillus subtilis for highly producing D-ribose and co-producing acetoin, which is preserved in a China general microbiological culture collection center with CGMCC No.3720 and the preservation data of April eighth, 2010. The invention also discloses a method for producing the D-ribose and co-producing the acetoin by fermenting the bacillus subtilis. The selected bacterial strains can produce produce the D-ribose and co-produce the acetoin by using the fermentation of a plurality of types of carbon and nitrogen sources, the operation is convenient and simple, the conditions of culture are extensive, and the industrialization is easy. In the invention, an organic acid is added to a fermentation medium, therefore, the yield of the D-ribose is improved by 64.5g / L compared with the original strains, and the acetoin is improved by 17.3g / L. The bacillus subtilis has higher capability for synthetizing the D-ribose and the acetoin, and the yield of reduced by-products of 2,3-butanediol of the acetoin is low.

Description

technical field [0001] The invention belongs to the technical field of microorganisms and fermentation engineering, and in particular relates to a bacillus subtilis capable of high-yielding D-ribose and co-producing acetoin. Background technique [0002] D-ribose is a five-carbon sugar with important physiological and biochemical functions. For the physiologically active substances ATP, RNA, NAD + , NADP, FAD and CoA, D-ribose in the form of ribose group is an important component. D-ribose is widely used in food, medical and pharmaceutical fields, and has huge economic value. D-ribose can be used to produce flavor enhancers (5'-nucleotides) and riboflavin (vitamin B 2 ), pharmaceutical preparations, feed, cosmetics and food additives. More recent research has focused on its chemical conversion into antiviral and anticancer drugs. New research finds that for patients with fibromyalgia and chronic fatigue, D-ribose is beneficial for mental calm and sleep maintenance. [0...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/26C12N13/00C12N15/01C12N1/20C12P19/02C12R1/125
Inventor 应汉杰柏建新房婷陈晓春熊健陈勇
Owner NANJING UNIV OF TECH
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