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A kind of thermophilic anaerobic bacterium and the method for using it to produce ethanol

A thermophilic anaerobic bacillus and ethanol technology, which is applied in the field of thermophilic anaerobic bacillus and its use in the production of ethanol, can solve the problems of increased production cost, long fermentation cycle, long growth lag period, etc., and achieve less by-products and process scale-up easy effect

Inactive Publication Date: 2017-02-01
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in the case of high-concentration sugar substrates, the strain is affected by high-sugar osmotic stress, which leads to a long growth lag period, a long fermentation cycle, and a substantial increase in production costs.

Method used

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  • A kind of thermophilic anaerobic bacterium and the method for using it to produce ethanol
  • A kind of thermophilic anaerobic bacterium and the method for using it to produce ethanol
  • A kind of thermophilic anaerobic bacterium and the method for using it to produce ethanol

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Experimental program
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Embodiment 1

[0025] Acclimatization and isolation of strains

[0026] High sugar resistant thermophilic anaerobic bacteria strain of the present invention can be obtained like this:

[0027] In the present invention, the starting strain used for screening is Thermoanaerobacterium aotearoense SCUT27 / Δldh, which was screened, mutated and preserved in the laboratory, which is denoted as P8G0 here.

[0028] Screening medium: Glucose: 80-100g·L -1 ;Xylose: 40-60g·L -1 ; (NH 4 ) 2 SO 4 : 1-5g·L -1 ; MgCl 2 ·6H 2 O: 1-5g·L -1 ;KH 2 PO4 : 0.5-5g·L -1 ;K 2 HPO 4 : 0.5-5g·L -1 ; CaCl 2H 2 O: 0.1-2.0g·L -1 ;Na-β-glycerophosphate: 2-10g·L -1 ;FeSO 4 ·7H 2 O: 0.0001-0.001g·L -1 ;Yeast extract: 2-10g·L -1 ; Cysteine ​​hydrochloride monohydrate (C 3 h 7 NO 2 S·HCl·H 2 O): 0.1-2.0g·L -1 ; Resazurin: 0.001-0.01g L -1 ;Agar powder: 5-20g·L -1 ; Erythromycin: 20-100ng / μl.

[0029] The starting strain P8G0 was first cultured in an anaerobic shake flask with 81g / L screening medium (...

Embodiment 2

[0034] Identification of P8G3#4 strains after domestication

[0035] In order to identify the differences of the strains before and after domestication, transcriptomics method was used to analyze the differences of metabolic genes between P8G0 and P8G3#4, and cluster analysis was performed on the differential genes to identify the gene expression levels that lead to the differences in the growth and metabolism of the two strains of P8G0 and P8G3#4 s reason.

[0036] Transcriptome sequencing analysis was performed on the starting strain Thermoanaerobacterium aotearoense SCUT27 / Δldh P8G0 and the screened mutant strain P8G3#4 that can grow normally under high concentration of substrate. Analyze the differentially expressed genes in the two samples, the screening standard is |logFC|>=1 (ie two-fold difference), analyze the differentially expressed genes in the two samples, P8G0 and P8G3#4 have a total of 69 genes in the expression abundance There were significant differences. Com...

Embodiment 3

[0040] Shake flask fermentation culture to compare the growth difference between the domesticated high-sugar-tolerant strain and the original strain

[0041] Preparation of seed solution:

[0042] The seed medium adopts MTC improved medium, and various combinations of them can be divided into A, B, C, D, and E liquids, which are sterilized by high temperature respectively, and then mixed according to the ratio of 45:2:1:1:1 before use . The specific composition of each solution is shown in Table 1.

[0043]

[0044] Specifically, the preparation method of the seed culture medium is as follows: prepare liquids A, B, C, and D respectively and put them in serum bottles, vacuumize, fill with nitrogen, sterilize at 115°C for 20 minutes, and store them for later use; liquid E is sterilized by filtration and directly injected into Store in the serum bottle that has been filled with nitrogen and sterilized for later use; take the serum bottle that has been filled with liquid A, a...

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Abstract

The invention relates to the field of microbial strains, in particular to a thermophilic anaerobic bacillus and a method for producing ethanol by using it. The bacterial strain of the present invention is Thermoanaerobacterium aotearoense P8G3#4, which has been preserved in the General Microorganism Center of China Microbiological Culture Collection Management Committee, the preservation number is CGMCC NO.9000, and the preservation date is April 3, 2014 . The method of utilizing said bacterial strain to produce ethanol: first prepare the seed liquid of thermophilic anaerobic bacillus CGMCC9000, then transfer the thermophilic anaerobic bacillus CGMCC9000 seed liquid to the fermentation medium with 10-15% w / w inoculation amount, in Stirring culture under anaerobic conditions, and finally ethanol is separated from the fermentation broth. The strain has good genetic stability, stable yield traits, and the fermentation lag period is shortened to 1 / 4 of the original one, saving energy consumption.

Description

technical field [0001] An ethanol-producing bacterial strain and its domestication method for improving high-concentration sugar tolerance, and a method for producing ethanol using the bacterial strain. The invention relates to a method for producing ethanol by using glucose and xylose as main raw materials and utilizing a mutant strain of Thermoanaerobacterium aotearoense SCUT27 / Δldh to ferment. Background technique [0002] Ethanol is a new type of clean fuel and an important direction for the development and utilization of renewable energy. The development of bioethanol is the general trend. The second-generation bioethanol is produced by pretreatment of plant fibers, hydrolysis with inorganic acid or cellulase, and then bio-fermentation. Hydrogen energy has attracted the attention of researchers because of its cleanliness, high energy density, and various preparation methods. [0003] The main obstacle to the production of ethanol by fermentation of cellulosic substan...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P7/06C12R1/01
CPCC12P7/06C12N1/205C12R2001/01C12N1/20Y02E50/10
Inventor 李爽赖志城王菊芳
Owner SOUTH CHINA UNIV OF TECH
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