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Method for producing D-1,2,4-butantriol through bio-converting cellulosic hydrolyzate

A cellulose hydrolyzate and biotransformation technology, applied in the biological field, can solve the problem of low yield, and achieve the effects of improving purity, saving costs, and reducing bypass pathways.

Active Publication Date: 2016-11-23
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, domestic and foreign studies on butanetriol fermentation mostly focus on using xylose as raw material directly, and the yield is not high, generally 0.88-3.96g / L, and there is no report on using cellulose as raw material

Method used

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  • Method for producing D-1,2,4-butantriol through bio-converting cellulosic hydrolyzate
  • Method for producing D-1,2,4-butantriol through bio-converting cellulosic hydrolyzate

Examples

Experimental program
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Effect test

Embodiment 1

[0025] Example 1 Construction of genetically engineered bacteria

[0026] Construction of clones expressing 2-ketoacid decarboxylase (mdlC), D-xylose dehydrogenase (xylB), xylose dehydratase (yjhG) and alcohol dehydrogenase (adhP), knockout host bacteria for xylose utilization and D -BT synthesizes the xylose isomerase (xylA) gene in the intermediate metabolite decomposition pathway, and obtains genetically engineered bacteria, among which, 2-ketoacid decarboxylase (mdlC), GenBank: AY143338.1; D-xylose dehydrogenase (xylB), Gene ID: 7329904; Xylose Dehydratase (yjhG), Gene ID: 946829; Alcohol Dehydrogenase (adhP), Gene ID: 00 946036; Xylose Isomerase (xylA), Gene ID: 948141 .

Embodiment 2

[0027] Example 2 Effects of Different Fermentation Temperatures on the Production of Butanetriol Using Corncob Hydrolyzate as Substrate

[0028] The suspended hydrolyzate mixed with 2% (v / v) sulfuric acid and corn cob hydrolyzate at a ratio of 1:5 (w / v) was sterilized in an autoclave at 121°C for 20 minutes. Add alkaline reagent Ca(OH) 2 To neutralize sulfuric acid, the added Ca(OH) 2 The concentration is equal to the number of moles of the same sulfuric acid concentration, and NaOH is added to adjust the pH to 7.2. Finally, use filter paper to filter out the solid matter in the corn cob hydrolyzate pretreatment method, then add 2% (w / v) activated carbon to the corn cob hydrolyzate, heat at 50°C for 30 minutes, and then use filter paper to filter out the activated carbon to obtain clarification corncob hydrolyzate. The concentration of xylose in the obtained corncob hydrolyzate is 44g / L, add 10g / L Nacl, 5g / L yeast powder and 10g / L peptone in every liter of corncob hydrolyza...

Embodiment 3

[0029] Example 3 The impact of buffer solution on the production of butanetriol with corncob hydrolyzate as substrate

[0030] The suspended hydrolyzate mixed with 2% (v / v) sulfuric acid and corn cob hydrolyzate at a ratio of 1:5 (w / v) was sterilized in an autoclave at 121°C for 20 minutes. Add alkaline reagent Ca(OH) 2 To neutralize sulfuric acid, the added Ca(OH) 2 The concentration is equal to the number of moles of the same sulfuric acid concentration, and NaOH is added to adjust the pH to 7.2. Finally, use filter paper to filter out the solid matter in the corncob hydrolyzate, then add 2% (w / v) activated carbon to the corncob hydrolyzate, heat at 50°C for 30 minutes, and then filter out the activated carbon with filter paper to obtain clarified corncob hydrolyzate liquid. The concentration of xylose measured in the clarified corncob hydrolyzate is 44g / L, and 10g / L Nacl, 5g / L yeast powder and 10g / L peptone are added in every liter of corncob hydrolyzate, sterilized, and...

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Abstract

The invention discloses a method for producing D-1,2,4-butantriol through bio-converting cellulosic hydrolyzate. The method comprises the steps of constructing genes for cloning and expressing 2-keto acid decarboxylase, D-xylitol dehydrogenase, D-xylonic acid dehydrase and D-alcohol dehydrogenase, shifting the constructed genes into cells of host bacteria for knocking out xylose isomerase so as to obtain genetic engineering strains, culturing the genetic engineering strains, inoculating the genetic engineering strains to the cellulosic hydrolyzate, and carrying out fermentation so as to produce the D-1,2,4-butantriol. The method disclosed by the invention is easy and feasible, is high in yield and is applicable to industrialization.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for producing D-1,2,4-butanetriol by biotransforming cellulose hydrolyzate. Background technique [0002] D-1,2,4-Butanetriol is an important unnatural polyol, an important substrate in the synthesis of many natural products, and a synthetic precursor of many chiral compounds. In the military, D-1,2,4-butanetriol can be used to synthesize butanetriol trinitrate (BTTN) as rocket propellant. In medicine, D-1,2,4-butanetriol can be used to prepare the cholesterol-lowering drug Movinolin, the anticancer drug compatin, the drug hydroxyeicosatetraenoic acid (12-HETE) for the treatment of skin diseases and the AIDS drug3 -Hydroxy-tetrahydrofuran, etc. [0003] At present, the commercial production of 1,2,4-butanetriol mostly adopts chemical synthesis methods, such as Adkins et al. reported the use of malic acid reduction method, using different catalysts (Cu-Cr, Cu-Al, Ru-Re), at 2...

Claims

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Application Information

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IPC IPC(8): C12P7/18C12R1/19
CPCC12N9/0006C12N9/88C12N9/92C12P7/18C12Y401/01C12Y402/01082C12Y503/01005C12P2201/00
Inventor 陈可泉刘静王昕陈西欧阳平凯
Owner NANJING UNIV OF TECH
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