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Screening and fermentation method for producing 2,3-butanediol strains by using straws

A butanediol and straw technology, applied in fermentation, bacteria, etc., can solve the problem of high cost of cellulase

Inactive Publication Date: 2011-02-09
INST OF PROCESS ENG CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, one of the obstacles to the industrialization of lignocellulose fermentation chemicals is that the cost of cellulase is too high, generally accounting for about 50% of the total production cost. The enzyme can produce 2,3-butanediol strain at the same time, use straw as raw material to ferment 2,3-butanediol under the condition of not adding cellulase, and it is a cheap 2,3-butanediol production provides an efficient way

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] Take 3ml from the activated sludge sample, put it into a conical flask filled with 100ml sterile water and glass beads, vibrate vigorously on a shaker for 20min, and then keep it in a water bath at 90°C for 20min. Then serially diluted with sterile water (10 -3 , 10 -4 , 10 -5 , 10 -6 , 10 -7 ), each took 0.2ml, spread it on the medium with microcrystalline cellulose as the only carbon source, cultivated it in a 37°C incubator for 40-48h, and picked a single colony on the Congo red microcrystalline cellulose medium ( L -1 :KH 2 PO 4 0.5, MgSO 4 0.25, microcrystalline cellulose powder 1.88, Congo red 0.2, agar 20, gelatin 2.0, pH 7.0) after streak culture for 48 hours, pick the colony with obvious growth, and after purification, the bacteria with cellulase secretion ability can be obtained kind.

Embodiment 2

[0014] By methyl red experiment (M.R. experiment) and acetylmethylmethanol experiment (V.P. experiment) preliminary judgment produces the bacterial strain of 2,3-butanediol: the bacterium bacterial strain that will isolate from embodiment 1 is inoculated in peptone water culture medium ( L -1 : Peptone 3; Glucose 5; NaCl 5; pH natural), cultured at 35°C for 48 hours, sampling observation: Take 1ml culture solution in a test tube, add a drop of methyl red reagent and shake well, if the strain metabolizes If acid is produced, the fermentation liquid will be bright / orange red, indicating that the methyl red test is positive. If the strain does not produce acid during metabolism, and the fermentation liquid is nearly neutral, it will be yellow, indicating that the methyl red test is negative.

[0015] Take another 2ml of culture solution in another test tube, add 0.7-0.8ml of 40% KOH solution and 0.7ml of 6% α-naphthol alcohol solution after strong shaking, observe within 3 minute...

Embodiment 3

[0018] With straw as the carbon source, from the strains screened in Example 2, select excellent strains that can produce 2,3-butanediol and simultaneously secrete cellulase, and put 25ml of steam-exploded straw as the carbon source in a 100ml Erlenmeyer flask. Medium (g L -1 : peptone 3; glucose 20; NaCl 5; pH natural). Before starting the fermentation, pick 2 rings of different strains and insert them into the culture medium, and place them under the conditions of 35°C and 100-150r / min for shaking culture. During the cultivation process, samples were taken regularly to detect the production of 2,3-butanediol in the fermentation broth. The strain YB-4 with the highest yield of 2,3-butanediol was screened out, and was deposited in the General Microorganism Center of China Committee for the Collection of Microbial Cultures on July 14, 2010, with the preservation number: CGMCC No.4005.

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Abstract

The invention belongs to separation and screening of strains and high-value utilization field of straw resources, and particularly relates to a screening method for producing 2,3-butanediol and cellulase strains at the same time and a method for producing 2,3-butanediol by direct fermentation by using straws as a raw material. The invention first provides the method for screening strains capable of producing the 2,3-butanediol and secreting cellulase at the same time from active sludge, the strains YB-4 capable of producing the 2,3-butanediol and secreting the cellulase at the same time are obtained by screening, the strains YB-4 are preserved in CGMCC on July 14th, 2010, and the preservation number is CGMCC No. 4005. Meanwhile, the invention provides the method for producing the 2,3-butanediol by using the fermentation straws of the strains, wherein the strains take the steam explosion straws as a carbon source, a nitrogen source is cultured for 4 to 7 days in a fermentation medium to obtain yeast extract, and the yield of the 2,3-butanediol is 8 to 12 percent of the dry weight of the straws (2,3-butanediol / straw dry weight, W / W). The method for producing the 2,3-butanediol by using the straws as the raw material does not need adding cellulase so as to greatly reduce the production cost.

Description

technical field [0001] The invention belongs to the field of separation and screening of excellent strains and high-value utilization of straw, and in particular relates to a screening method for strains capable of producing 2,3-butanediol and cellulase at the same time and directly fermenting and producing 2,3-butane with straw as raw material diol method. Background technique [0002] 2,3-Butanediol is a colorless, odorless chiral compound with three stereoisomers: dextro-, levo-, and meso-isomers. 2.3-Butanediol has a molecular weight of 90.12kDa, a higher boiling point (180-184°C), and a lower freezing point (-60°C). As an extremely valuable liquid fuel, its combustion value is 27,198J g -1 , can be combined with methanol (22,081J g -1 ) and ethanol (29,005J g -1 )Comparable. It can generate methyl ethyl ketone, diacetyl, butene and butadiene through dehydration, dehydrogenation and hydration reactions. [0003] As a chemical intermediate, 2,3-butanediol can be use...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/18C12N1/20
Inventor 陈洪章彭小伟
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
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