Bacillus and application thereof to production of acetoin and 2, 3- butanediol through high temperature fermentation

A Bacillus, high temperature fermentation technology, applied in the biological field, can solve the problems of high cost, unsuitable for low cost, large-scale industrial production and the like

Active Publication Date: 2016-08-03
CHINA UNIV OF PETROLEUM (EAST CHINA)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] So far, there are very few research reports on the production of acetoin and 2,3-butanediol by high-temperature fermentation at home and abroad. The research group of the inventor once used a strain of Geobacillus bacteria to produce these two products by high-temperature fermentat

Method used

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  • Bacillus and application thereof to production of acetoin and 2, 3- butanediol through high temperature fermentation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Strain enrichment screening and isolation and purification

[0030] 1.1 Environmental sample collection

[0031] Samples were collected from Jiaozhou Bay Beach and Tangdao Bay Park in Qingdao City, Shandong Province.

[0032] 1.2 Initial screening of strains

[0033] The above samples were added to sterile water, stirred rapidly and naturally settled for 10 minutes, then the supernatant was diluted appropriately and spread on Luria-Bertani solid medium, and incubated at 60°C for 24 hours. Pick a single colony, separate and purify by streaking multiple times to obtain multiple pure strains.

[0034] 1.3 Strain re-screening

[0035] The pure strains obtained in the above experiments were inoculated into the re-screening medium respectively, and after cultured in shake flasks at 55° C. for 24 hours, the products were detected by gas chromatography. Each liter of the re-screening medium contains: 10g of peptone, 5g of yeast powder, 10g of sodium chloride, and...

Embodiment 2

[0037] Embodiment 2: strain identification and preservation

[0038] The bacterial strain H15-1 obtained in Example 1 was tested, and its Gram staining was positive, spores were produced, and the cell shape was rod-shaped; under the condition of 55 ° C, it was cultured with Luria-Bertani solid medium for 6 hours, and the diameter could be obtained. Gray-white colonies about 2-3mm in size with folds on the surface. The bacterial strain H15-1 of the present invention can grow in the range of 40-60°C, and proliferates fastest at 50°C, indicating that the bacterial strain is a thermophilic strain.

[0039] By sequencing the 16SrDNA of the bacterial strain of the present invention, and comparing the 16SrDNA sequence of the bacterial strain of the present invention with the 16SrDNA sequence of the bacterial model strains recorded in the EzTaxon database, it was found that the 16SrDNA sequence of the bacterial strain of the present invention is consistent with the existing The homol...

Embodiment 3

[0041] Embodiment 3: The application method of producing acetoin and 2,3-butanediol by high-temperature fermentation of Bacillus sp.H15-1

[0042] The specific implementation steps are as follows:

[0043] 3.1 Activated strains

[0044] Streak inoculation of the strain Bacillus sp.H15-1 provided by the present invention onto the Luria-Bertani solid medium, and culture in a thermostat at 55° C. for more than 6 hours until the colony grows plump.

[0045] The Luria-Bertani solid medium contains per liter: 10g peptone, 5g yeast powder, 10g sodium chloride, and 17g agar powder. Adjust the pH of the medium to 7.0.

[0046] 3.2 Preparation of liquid seeds

[0047] The colonies obtained in the above steps were picked with an inoculation loop, inoculated into Luria-Bertani liquid medium, and cultured on a shaker at 50° C. for 10 h. A baffle shaker flask with a volume of 250 mL is used, and the liquid volume is 50 mL; a rotary shaker is used, and the rotation speed is set at 150 rp...

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Abstract

The invention discloses bacillus and application thereof to production of acetoin and 2, 3- butanediol through high temperature fermentation. A strain is Bacillus sp. H15-1CGMCC No. 12389, the gram faerbung is positive, spores are produced, the cellular morphology is in a rod shape, and the stain can grow at the temperature of 40-60 DEG C and proliferate at the highest speed at the temperature of 50 DEG C. By means of the strain, degerminated corn flour hydrolysate is adopted as the main raw material, a mechanical stirring and ventilation fermentation tank is adopted for conducting fermentation for 68 hours at the temperature of 50 DEG C, and acetoin with the concentration being 50.8 g/L and 2, 3- butanediol with the concentration being 32.1 g/L can be generated. A method for producing acetoin and 2, 3- butanediol through high temperature fermentation has the advantages that raw materials are low in cost, the fermentation process is resistant to infectious microbe contamination, cooling water is saved, and operation is conducted coarsely.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a bacillus strain and its application. Background technique [0002] Acetoin is 3-hydroxy-2-butanone, which has important uses in food, fragrance, cosmetics, chiral synthesis and other fields (Xiao and Lu, 2014). 2,3-Butanediol has important uses in chemical synthesis, food, fuel, antifreeze and other fields (Jie et al., 2011). Acetoin and 2,3-butanediol can be fermented from renewable raw materials and are important emerging platform compounds (Xue et al., 2012). The two are the products of the acetoin metabolic pathway in some bacteria, and can be transformed into each other through the action of 2,3-butanediol dehydrogenase in the cell. Acetoin and 2,3-butanediol often exist together in fermentation products, but their boiling points are quite different, and they can be easily separated from each other by distillation. [0003] The production of acetoin and 2,3-buta...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P7/26C12P7/18C12R1/07
CPCC12P7/18C12P7/26C12N1/205C12R2001/07
Inventor 肖梓军顾如林侯孝元吕新赵静宜
Owner CHINA UNIV OF PETROLEUM (EAST CHINA)
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