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Method of synthesizing poly 3-hydracrylic acid by utilizing recombinant Escherichia coli

A technology of recombining Escherichia coli and hydroxypropionic acid, which is applied in the field of genetic engineering, can solve the problems of NAD+ deficiency, affecting the synthesis of P3HP, etc., and achieve the effects of reducing loss rate, balancing reducing power, and maintaining stability

Inactive Publication Date: 2015-04-01
天津申宗科技有限公司
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AI Technical Summary

Benefits of technology

This patented method allows for increased production of pigmented acid (PA) without consuming too much or losing important enzymes like phosphatases during cell growth. It also introduces an intermediate product called 2,3-pDPG, which helps keep it stable when being used instead of producing PA from other compounds such as glucose. Overall this new technique improves efficiency and yield of chemical processes involved in making certain types of molecules.

Problems solved by technology

Technological Problem: Current methods for making poly(3- hydroxy propional acid)) require costly starting materials like beta lactic acid or other similar ones. Additionally, current techniques involve complicated processes involving multiple steps leading to low yields and poor quality products.

Method used

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  • Method of synthesizing poly 3-hydracrylic acid by utilizing recombinant Escherichia coli
  • Method of synthesizing poly 3-hydracrylic acid by utilizing recombinant Escherichia coli
  • Method of synthesizing poly 3-hydracrylic acid by utilizing recombinant Escherichia coli

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Embodiment 1

[0027] A recombinant Escherichia coli strain, which integrates the synthetic gene of glycerol dehydratase and its activator, and the dhaT gene on the Escherichia coli genome, and the recombinant Escherichia coli strain has the ability to biosynthesize poly-3-hydroxypropionic acid and 1,3-propanediol ability.

[0028] The dhaT gene is obtained by cloning Klebsiella pneumoniae genome DNA with primers ZG-440 and ZG-441 as a template.

[0029] A construction method utilizing said recombinant escherichia coli strain, the steps are as follows:

[0030] Step 1: Construction of plasmid pET21 (A): using Klebsiella pneumoniae genomic DNA as a template, using primers ZG-440 and ZG-441 to clone the dhaT gene, the dhaT fragment and plasmid pWQ02 were digested with HindIII and XhoI and then ligated;

[0031] Step 2: Construction of plasmid pRE112(B): Using the genome of E.coliBL21 as a template, use primers ZG-268 and ZG-269, primers ZG-270 and ZG-271 to amplify the homologous fragments on...

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Abstract

The invention relates to a method of synthesizing poly 3-hydracrylic acid by utilizing recombinant Escherichia coli and protects a recombinant Escherichia coli strain, a construction method of the recombinant Escherichia coli strain and a method of synthesizing the poly 3-hydracrylic acid by using the recombinant Escherichia coli strain by virtue of a biological method. The method has the beneficial effects that in the metabolic pathway of biologic synthesis of P3HP with glycerol serving as a substrate, NAD+, serving as coenzyme, can generate a great deal of NADH when used for catalyzing 3-hydroxy-propionaldehyde to produce 3HP; the oxidation-reduction disequilibrium in a recombinant strain, caused by increase of NADH is capable of inhibiting further increase of the yield of P3HP, and NADH can be consumed in the process that the recombinant Escherichia coli strain metabolizes glycerol to generate 1,3-PDO, and therefore synthesis of 1,3-PDO is introduced into the synthesis pathway of P3HP, the in-vivo reducing power of the strain can be balanced, the plasmid loss rate is effectively reduced, and the plasmid stability is kept.

Description

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Claims

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Application Information

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Owner 天津申宗科技有限公司
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