Method for producing 2, 3-butanediol and special bacillus licheniformis thereof

A technology of bacillus licheniformis and butanediol, which is applied in the direction of microorganism-based methods, biochemical equipment and methods, bacteria, etc., can solve the problems of unseen bacillus licheniformis production, achieve high production intensity, reduce energy consumption, The effect of high yield

Active Publication Date: 2012-03-28
上海肆芃科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] After searching, there is no report on the method of producing 2,3-butanediol using thermophilic Bacillus licheniformis

Method used

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  • Method for producing 2, 3-butanediol and special bacillus licheniformis thereof
  • Method for producing 2, 3-butanediol and special bacillus licheniformis thereof

Examples

Experimental program
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Embodiment 1

[0029] Example 1: Screening of high sugar tolerance and capable of producing 2,3-butanediol strains

[0030] Soil samples were obtained from orchards and other places, soaked in screening medium for 2 hours, filled with 50 mL of soaking solution in a 500 mL Erlenmeyer flask, and cultured on a shaker at 180 r / min for 24 hours at 50 °C. Dilute the culture medium with 10 -2 、10 -3 、10 -4 、10 -5 、10 -6 Five dilutions were spread on petri dishes of solid screening medium, and cultured at 50°C for 12h. Pick a single colony, expand the culture, and conduct a preliminary screening through the VP reaction. Positive strains were screened out based on reaction time and color change. The screening principle is that 2,3-butanediol is oxidized to diacetyl under alkaline conditions, and the red substance is synthesized with creatine or guanidine derivatives. Adding α-naphthol and creatine can promote the reaction. Then measure the 2,3-butanediol production of the positive bacteria scr...

Embodiment 2

[0034] Embodiment 2: prepare the cell liquid culture of Bacillus licheniformis CCTCC M 2011371 bacterial strain

[0035]Pick a loop of Bacillus licheniformis CCTCC M 2011371 strain cultured on the slant of solid high-sugar LB medium, inoculate it in a 500mL Erlenmeyer flask containing 100mL of sterilized high-sugar LB medium, and place it in a shaker at 55°C. Cultivate on the bed at a rotational speed of 180 r / min for 24 hours to obtain a cell liquid culture of Bacillus licheniformis CCTCC M 2011371 strain.

[0036] The formula of the above-mentioned high-glucose LB medium is as follows: 1L of distilled water contains: 150g of glucose, 10g of peptone, 5g of yeast powder, 10g of NaCl, and sterilized at 115°C for 15min. The solid high-glucose LB medium is based on the above formula by adding 20g / L agar powder.

Embodiment 3

[0037] Embodiment 3: Utilize bacillus licheniformis CCTCC M 2011371 strain to ferment and produce 2,3-butanediol with fermentation medium 1 in Erlenmeyer flask

[0038] The liquid cell culture of the Bacillus licheniformis CCTCC M 2011371 bacterial strain obtained by Example 2 was inoculated in a 500mL Erlenmeyer flask containing sterilized 100mL fermentation medium with an inoculum size of 50mL / L, at 50°C, Place on a shaker and cultivate at a rotational speed of 180r / min. Samples are taken at intervals of 4 hours and the content of 2,3-butanediol is detected. At 34 hours, the concentration of 2,3-butanediol reaches 56.0g / L. At 60h, the Erlenmeyer flask was removed from the shaker to stop the fermentation.

[0039] Above-mentioned fermentation medium 1 formula is as follows:

[0040] Glucose 120 g / L, yeast powder 10 g / L, corn steep liquor 5 g / L, potassium dihydrogen phosphate 4 g / L, sodium acetate 1 g / L, potassium chloride 2 g / L.

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Abstract

The invention discloses bacillus licheniformis 5-2-D and a method for producing 2, 3-butanediol by using the same on the premise of taking the temperature of 42-55 DEG C and the culture stirring time of 20-60 hours as fermentation conditions and taking glucose as a substrate. In the method for producing the 2, 3-butanediol, which is provided by the invention, the advantages of high-temperature fermentation process are fully utilized, the probability of mixed fungi pollution is small, the sterilization is not performed on a fermentation culture medium, and the energy consumption and the loss of nutrient substances are reduced; simultaneously, the highest concentration of the obtained 2, 3-butanediol can achieve 92g / L and the production strength can achieve 2.56g / L / hour.

Description

technical field [0001] The invention relates to a method for producing 2,3-butanediol and its special bacillus licheniformis. Background technique [0002] 2,3-Butanediol is widely used in many fields such as chemical industry, food, fuel and aerospace (Appl Microbiol Biotechnol, 2001, 55(1): 10-18). Because of its high calorific value (27,200kJ / kg), which is equivalent to ethanol (29,100kJ / kg), it can be used as a fuel additive; it can replace 1,4-butanediol and be used in the synthesis of polyester and polyurethane ; Its dehydration product methyl ethyl ketone is a low boiling point solvent, widely used in coatings, lubricants, fuels and other industries, and can be used as an intermediate for organic synthesis of spices, antioxidants, etc.; its dehydration product 1,3-butadiene can be used It is an important basic organic raw material for petrochemical industry; it can be condensed and hydrogenated with methyl ethyl ketone to form octane, which can be used to produce hig...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P7/18C12R1/10
Inventor 马翠卿李理想张丽杰许平
Owner 上海肆芃科技有限公司
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