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Application of Bacillus subtilis in high-yield adenosine and acetoin co-production

A technology of Bacillus subtilis and acetoin, which is applied in the field of high-yield adenosine and co-production of acetoin of Bacillus subtilis, can solve the problems of high cost and low utilization rate of raw materials, and achieve high-yield, stable traits, and cultivation Effects of extensive conditions, high ability to synthesize adenosine and acetoin

Inactive Publication Date: 2013-01-16
NANJING UNIV OF TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In summary, the existing technologies all use Bacillus subtilis to produce adenosine or acetoin alone, which has the problems of low utilization rate of raw materials and high cost

Method used

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  • Application of Bacillus subtilis in high-yield adenosine and acetoin co-production

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Slant medium: glucose 15g / L, peptone 10g / L, yeast extract 5g / L, corn steep liquor 10ml / L, MgSO 4 ·7H 2 O 0.1g / L, NaCl 3g / L, guanosine 0.03g / L, agar 15g / L, the rest is water, pH 7.0;

[0035] Seed medium: glucose 15g / L, peptone 10g / L, yeast extract 10g / L, corn steep liquor 10ml / L, MgSO 4 ·7H 2 O 0.1g / L, NaCl 4g / L, guanosine 0.03g / L, the rest is water, pH 7.0;

[0036] Fermentation medium: glucose 80g / L, yeast extract 15g / L, (NH 4 ) 2 SO 4 16g / L, urea 3g / L, KH 2 PO 4 4g / L, KCl 4g / L, MgSO 4 ·7H 2 O 0.5g / L, MnSO 4 ·H 2 O 0.01g / L, soybean cake hydrolysate 20ml / L, CaCO 3 30g / L, the rest is water, pH7.0;

[0037] The strain A409 was transferred from the inclined plane to a ring in a 500 mL shake flask containing 30 mL of liquid seed culture medium, and cultured on a reciprocating shaker at 200 rpm at 32° C. for 20 hours. Transfer 3 mL to a 500 mL shake flask containing 30 mL of liquid fermentation medium, and ferment and culture for 66 hours at 33° C. on a 250 rpm shaker. The outp...

Embodiment 2

[0039] Fermentation medium: glucose 100g / L, yeast extract 15g / L, (NH 4 ) 2 SO 4 16g / L, urea 3g / L, KH 2 PO 4 4g / L, KCl 4g / L, MgSO 4 ·7H 2 O 0.5g / L, MnSO 4 ·H 2 O 0.01g / L, soybean cake hydrolysate 20ml / L, CaCO 3 30g / L, the rest is water, pH7.0;

[0040] The strain A409 was transferred from the inclined plane to a ring in a 500 mL shake flask containing 30 mL of liquid seed culture medium (same as in Example 1), and cultured on a reciprocating shaker at 200 rpm at 32° C. for 20 hours. Transfer 2 mL to a 500 mL shake flask containing 20 mL of liquid fermentation medium, and ferment and culture at 33° C. on a 250 rpm shaker for 66 hours. The output of adenosine reached 21.6g / L, and the output of acetoin reached 13.6g / L.

Embodiment 3

[0042] Use Bacillus subtilis A409 to synthesize adenosine in a 5L fermentor to prepare a seed culture medium. The formula and culture method are the same as those in Example 1. The fermentation medium is prepared. The formula and culture method are the same as those in Example 2. 10% (v / v) of the inoculum is connected to the cooled fermentation broth, the volume of the fermentation medium is 3L, the aeration is controlled to 2vvm, the stirring speed is 500rpm, the pH is adjusted to 6.5 with 20% ammonia water, 35℃ After 66 hours of fermentation, the content of adenosine in the final fermentation broth is 23g / L and the content of acetoin is 15g / L;

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Abstract

The invention discloses application of Bacillus subtilis in high-yield adenosine and acetoin co-production. The Bacillus subtilis is CGMCC No. 4484. Screened strains can be fermented for adenosine production and acetoin co-production by multiple carbon sources and nitrogen sources. Yield of adenosine and acetoin can be evidently increased by adding sodium citrate into fermentation medium. Operation is convenient and simple, culture conditions are extensive, a higher level can be kept on a 5L fermentation tank, and potential of large-scale industrialization is provided.

Description

Technical field [0001] The invention belongs to the technical field of microorganisms and fermentation engineering, and specifically relates to the application of Bacillus subtilis in high-producing adenosine and co-producing acetoin. Background technique [0002] Adenosine, also known as adenosine, is composed of glycosidic bonds connecting adenine and ribose. It is an endogenous nucleoside with important physiological functions in human cells. Adenosine is well-known for its vasodilator effect. It has a wide range of cardiac effects and rapid and significant coronary dilatation. A large number of studies have shown that adenosine can also trigger or mediate ischemic preconditioning and reduce reperfusion injury. Protection effect. In the United States, adenosine is an FDA-approved first-line drug for the conversion of paroxysmal supraventricular tachycardia (PSVT), and it is also one of the two drugs approved by the FDA for cardiac drug stress testing. It has become an emergen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/40C12P7/26C12R1/125
Inventor 应汉杰柏建新张春伟陈晓春陈勇谢婧婧吴菁岚
Owner NANJING UNIV OF TECH
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