Recombinant escherichia coli for producing melatonin, and construction method and application of recombinant escherichia coli

A technology for recombining Escherichia coli and Escherichia coli, applied in the field of genetic engineering, can solve the problems of low expression, low expression, and low protein specific activity.

Active Publication Date: 2020-06-30
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the biosynthetic pathway of melatonin is very clear, the proteins involved in it have low specific activities, and most of them are difficult to express in the existing

Method used

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  • Recombinant escherichia coli for producing melatonin, and construction method and application of recombinant escherichia coli
  • Recombinant escherichia coli for producing melatonin, and construction method and application of recombinant escherichia coli
  • Recombinant escherichia coli for producing melatonin, and construction method and application of recombinant escherichia coli

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Embodiment 1

[0142] The construction of the recombinant escherichia coli strain of embodiment 1 melatonin high yield

[0143] 1. PCR amplification of coding sequences of genes related to melatonin biosynthesis

[0144] Taking the phenylalanine hydroxylase gene (shown in the 319th-1209th positions of SEQ ID NO.1) of the synthetic Xanthomonas campestris as a template, PCR amplification is carried out with primers F1 and R1 to obtain a gene containing phenylalanine PCR amplification product X of acid hydroxylase gene (shown in positions 299-1232 of SEQ ID NO.1).

[0145] Using the synthetic 4a-hydroxytetrahydrobiopterin dehydratase (phhB) gene (shown in the 1234-1590th position of SEQID NO.1) derived from Pseudomonas aeruginosa as a template for PCR amplification with primers F3 and R3, A PCR amplification product P (represented by positions 1210-1614 of SEQ ID NO. 1) comprising the gene phhB was obtained.

[0146] Using the synthesized dihydromonophosphate reductase (folM) gene (shown in p...

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Abstract

The invention discloses recombinant escherichia coli for producing melatonin, and a construction method and application of the recombinant escherichia coli. The invention first discloses the recombinant escherichia coli, and compared with receptor escherichia coli, the recombinant escherichia coli has the advantages that the expression amount of a gene of a melatonin biosynthesis-related protein is increased, and/or the content of the melatonin biosynthesis-related protein is increased, and/or activity of the melatonin biosynthesis-related protein is increased, wherein the melatonin is selected from at least one of phenylalanine hydroxylase, dihydromonophosphate reductase, 4alpha-hydroxytetrahydrobiopterin dehydratase, aspartate transaminase family protein, N-acetyltransferase and caffeicacid-O-methyltransferase.and further discloses a construction method and application of the recombinant escherichia coli. The recombinant escherichia coli strain for producing melatonin has melatoninsynthesis efficiency is obviously higher than other existing strains, and has great significance for industrial production and large-scale application of melatonin.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a melatonin-producing recombinant Escherichia coli and its construction method and application. Background technique [0002] Melatonin (N-acetyl-5-methoxytryptamine, melatonin, C 13 N 2 h 16 o 2 ) belongs to tryptophan derivatives and is an important antioxidant in organisms. Melatonin was first discovered in the pineal gland of cattle and is considered to be an important neurohormone. Later research found that melatonin is distributed in various organs in the human body and plays different important functions. Now it has been widely used in medicine In the field of health care, it is used to treat symptoms such as insomnia and depression. In 2019, the global market demand for melatonin reached 1 billion US dollars. Melatonin synthesis pathways widely exist in natural organisms, and the classic synthesis pathway in cells is: tryptophan → 5-hydroxytryptophan → 5-...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12N15/70C12P17/10C12R1/19
CPCC12N9/0071C12N9/0004C12N9/88C12N9/1096C12N9/1029C12N9/1007C12Y402/01096C12Y206/01001C12N15/70C12P17/10
Inventor 董志扬张岩峰张山何永志张楠
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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