Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

L-phenylalanine producing strain and construction method thereof

A technology of phenylalanine and construction method, which is applied in the field of genetic engineering, can solve the problems of no acid production, OD value drop, etc., and achieve the effects of increasing production, solving OD drop, and increasing metabolic flow

Pending Publication Date: 2021-11-30
XINTAI JIAHE BIOTECH CO LTD +1
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The L-phenylalanine-producing bacteria constructed by the present invention can effectively solve the problems that the OD value of the existing L-Phe production strains just after the logarithmic phase drops sharply and does not produce acid, and greatly increases the production of L-Phe

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • L-phenylalanine producing strain and construction method thereof
  • L-phenylalanine producing strain and construction method thereof
  • L-phenylalanine producing strain and construction method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1: Codon optimization

[0042] The nucleotide sequence of the pheA gene obtained by the inventor from the existing database is shown in SEQ ID NO.1, and the amino acid sequence of the encoded chorismate mutase is shown in SEQ ID NO.2. The nucleotide sequence of the aroF gene is shown in SEQ ID NO.3. The nucleotide sequence of the aroB gene is shown in SEQ ID NO.4. The nucleotide sequence of the aroE gene is shown in SEQ ID NO.5.

[0043] In order to make the pheA gene, aroF, aroB gene and aroE gene better adapt to the prokaryotic expression system, the present invention optimizes the codons of the above genes respectively.

[0044] The nucleotide sequence of the aroB gene after codon optimization is shown in SEQ ID NO.6; the codon relative fitness figure before optimization is shown in figure 1 Shown; the codon relative fitness map after optimization is shown in figure 2 shown.

[0045] The nucleotide sequence of the aroE gene after codon optimization is s...

Embodiment 2

[0048] Embodiment 2: Construction of the first recombinant expression vector

[0049] Plasmid pET-28a(+) was digested with NcoI and SacI, and then the aroB gene (shown in SEQ ID NO.6) was integrated into the double-digested plasmid pET-28a(+) to obtain a recombinant plasmid pET-aroB; then digest the recombinant plasmid pET-aroB with EagI and XhoI, and then integrate the aroE gene (shown in SEQ ID NO.7) into the digested recombinant plasmid pET-aroB to obtain the first Recombinant expression vector (pET-aroB-aroE).

[0050] The constructed first recombinant expression vector was digested with four enzymes NaoI, SacI, EagI and XhoI, and the results were as follows: Figure 9 shown. The results showed that the aroB gene (shown in SEQ ID NO.6) and aroE gene (shown in SEQ ID NO.7) had been successfully integrated into the plasmid pET-28a(+).

Embodiment 3

[0051] Embodiment 3: Construction of the second recombinant expression vector

[0052] Plasmid pGEX-2T was double-digested with BamHI and EcoRI, and then the aroF gene (shown in SEQ ID NO.8) was integrated into the double-digested plasmid pGEX-2T to obtain recombinant plasmid pGEX-2T-aroF, The recombinant plasmid pGEX-2T-aroF was digested with TthllllⅠ and AatⅡ, and the pheA gene (shown in SEQ ID NO.10) was integrated into the digested recombinant plasmid pGEX-2T-aroF to obtain the second Recombinant expression vector (pGEX-2T-aroF-PheA).

[0053] The constructed second recombinant expression vector was digested with four enzymes TthllllⅠ, AatⅡ, BamHI and EcoRI for verification, the results are as follows: Figure 10 shown. The results showed that the aroF gene (shown in SEQ ID NO.8) and pheA gene (shown in SEQ ID NO.10) had been successfully integrated into the plasmid pGEX-2T.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
wavelengthaaaaaaaaaa
Login to View More

Abstract

The invention discloses an L-phenylalanine producing strain and a construction method thereof. The L-phenylalanine producing strain contains a first recombinant expression vector and a second recombinant expression vector, wherein the first recombinant expression vector is used for expressing an aroB gene and an aroE gene in series; and the second recombinant expression vector is used for expressing an aroF gene and a pheA gene in series. By adopting the L-phenylalanine producing strain constructed by the invention, the problems that the OD value of an existing L-Phe producing strain is greatly reduced and acid is not produced just in a logarithmic phase can be effectively solved, and the yield of L-Phe is greatly improved.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to an L-phenylalanine-producing bacterium and a construction method thereof. Background technique [0002] L-Phenylalanine (L-Phe) is an important intermediate of pharmaceuticals and food chemicals. It participates in the synthesis of important neurotransmitters and hormones in animals, and is widely used in the fields of medicine and food. As a carrier of anticancer drugs, L-Phe can introduce anticancer drugs into the tumor site, and can greatly reduce the toxic and side effects of drugs while inhibiting tumor growth. In addition, L-Phe is also the main synthetic raw material for the low-calorie sweetener aspartame. With the increasing market demand for tumor drugs and sweetener aspartame, the market demand for L-Phe is also increasing year by year, which puts forward higher requirements for the industrial production of L-Phe. [0003] The methods for preparing L-Phe ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N15/70C12N15/53C12N15/54C12N15/60C12N15/61C12N15/66C12P13/22C12R1/19
CPCC12N9/88C12N9/0006C12N9/1085C12N9/90C12N15/70C12N15/66C12P13/222C12Y402/03004C12Y101/01025C12Y205/01054C12Y504/99005C12N2800/22C12N2800/101
Inventor 岳明瑞谢沛曹华杰郭永胜
Owner XINTAI JIAHE BIOTECH CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com