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Genetic engineering strain capable of coproducing isoprene and 1,3-propylene glycol and establishment method and application thereof

A technology of genetically engineered bacteria and isoprene, applied in the field of genetic engineering, can solve the problems of waste and loss of biosynthetic energy, and achieve the effects of improving productivity and balancing metabolism

Active Publication Date: 2017-01-25
QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] Theoretically, in this method, 2 molecules of NADPH out of 6 molecules of NADPH are used for the synthesis of isoprene, and 4 molecules of NADPH are left. This part of excess reducing power will be lost from the respiratory chain in addition to being used for the synthesis of other substances in the cell. , which is a waste of energy for biosynthesis

Method used

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  • Genetic engineering strain capable of coproducing isoprene and 1,3-propylene glycol and establishment method and application thereof
  • Genetic engineering strain capable of coproducing isoprene and 1,3-propylene glycol and establishment method and application thereof
  • Genetic engineering strain capable of coproducing isoprene and 1,3-propylene glycol and establishment method and application thereof

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Experimental program
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Embodiment 1

[0088] The construction of embodiment 1 genetically engineered bacteria

[0089] Overexpression of acetyl-CoA acyltransferase / HMG-CoA reductase gene and HMG-CoA synthetase in Escherichia coli in which the glycerol dehydratase gene and glycerol dehydratase reactivator gene were integrated in the genome and the glycerol kinase gene was knocked out gene, mevalonate kinase gene, phosphomevalonate kinase gene, pyrophosphomevalonate decarboxylase gene, isopentenyl pyrophosphate isomerase gene, isoprene synthase gene, aldehyde reductase gene and Transhydrogenase gene, the obtained genetically engineered bacteria can use glucose and glycerol mixed carbon source fermentation to co-produce isoprene and 1,3-propanediol, and at the same time realize the regeneration of NADPH in the cell. Metabolic pathways and cofactor modification methods of co-production strains such as figure 1 shown.

[0090] 1. Gene integration

[0091] 1.1 Integrate glycerol dehydratase gene dhaB and glycerol deh...

Embodiment 2

[0140] Example 2 Fermentative production of isoprene and 1,3-propanediol

[0141] 1. Seed medium: yeast powder 5g / L, NaCl 10g / L, peptone 10g / L. The culture of G01, G02, G03 and 2369 needs to add 100 μg / mL of ampicillin and 34 μg / mL of chloramphenicol, and only 34 μg / mL of chloramphenicol is added for the culture of G04 strain.

[0142] 2. Fermentation medium: K 2 HPO 4 ·3H 2 O 9.8g / L, Citric acid·H 2 O 2.1g / L, ferric ammonium citrate 0.3g / L, beef extract 5g / L, 1mol / L MgSO 4 ·7H 2 O 2ml / L, 1mL 1000×trace elements ((NH 4 ) 6 Mo 7 o 24 4H 2 O3.7g / L; ZnSO 4 ·7H 2 O 2.9g / L; H 3 BO 3 24.7g / L; CuSO 4 ·5H 2 O 2.5g / L; MnCl 2 4H 2 O 15.8g / L), the culture of G01, G02, G03 and 2369 needs to add 100 μg / mL of ampicillin and 34 μg / mL of chloramphenicol, and only 34 μg / mL of chloramphenicol is added for the culture of G04 strain. The addition of carbon sources glucose and glycerol in the fermentation medium was added according to the culture requirements with a mass volume c...

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Abstract

The invention discloses a genetic engineering strain capable of coproducing isoprene and 1,3-propylene glycol and an establishment method and application thereof, and belongs to the technical field of genetic engineering. In the genetic engineering strain, a glycerol dehydratase gene and a glycerol dehydratase gene reactivating enzyme gene are integrated, a glycerol kinase gene is knocked out, and HMG-CoA reductase, HMG-CoA synthetase, mevalonate kinase, phosphomevalonate kinase, pyrophosphomevalonate decarboxylase, IPP isomerase, isoprene synthetase, aldehyde reductase and transhydrogenase are overexpressed. According to the invention, through genetic engineering, a metabolic way capable of coproducing the isoprene and the 1,3-propylene glycol is successfully established in escherichia coli; by the establishment method, redox cofactors in cells of the genetic engineering strain are metabolized in a balanced way, and the yield in coproducing the isoprene and the 1,3-propylene glycol through glucose and glycerol fermentation is increased.

Description

technical field [0001] The invention relates to a genetically engineered bacterium for co-producing isoprene and 1,3-propanediol, its construction method and application, and belongs to the technical field of genetic engineering. [0002] technical background [0003] Isoprene (2-methyl-1,3-butadiene) is currently recognized as an important chemical platform compound with great commercial application value. Isoprene is the main monomer of synthetic rubber, and its consumption accounts for 95% of the total output of isoprene. In addition, isoprene is also widely used in pharmaceutical or chemical intermediates, food, adhesives and aviation fuels and other fields. At present, the preparation methods of isoprene mainly include petroleum-based raw material isopentane, isopentene dehydrogenation method, chemical synthesis method (including propylene dimerization method, acetylene-acetone method, isobutylene-formaldehyde method) and cracking C5 fraction extraction. distillation e...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12P5/02C12P7/18C12R1/19
CPCC12N9/0006C12N9/0036C12N9/1025C12N9/1029C12N9/1205C12N9/1229C12N9/88C12N9/90C12P5/026C12P7/18C12Y101/01002C12Y101/01034C12Y203/01009C12Y203/0301C12Y207/0103C12Y207/01036C12Y207/04002C12Y401/01033C12Y402/0103C12Y402/03027C12Y503/03002
Inventor 咸漠刘会洲郭静曹玉锦
Owner QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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