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Streptomyces griseorubens pectinesterase as well as coding gene and application thereof

A technology of pectin esterase coding and Streptomyces grisea, which is applied in the fields of genetic engineering and modern enzyme engineering, can solve the problems of few examples, achieve strong tolerance, high-efficiency expression, and improve stability

Inactive Publication Date: 2017-05-10
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Pectin esterase has a wide range of sources, and bacteria, fungi and actinomycetes can produce pectin esterase, but there are few examples of pectin esterase that can be produced and successfully applied to industrial production

Method used

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  • Streptomyces griseorubens pectinesterase as well as coding gene and application thereof
  • Streptomyces griseorubens pectinesterase as well as coding gene and application thereof
  • Streptomyces griseorubens pectinesterase as well as coding gene and application thereof

Examples

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Embodiment Construction

[0038] Below in conjunction with specific embodiment, further illustrate the present invention. It should be understood that these examples are only used to illustrate the present invention, not to limit the protection scope of the present invention. Improvements and adjustments made by those skilled in the art according to the present invention in practical applications still belong to the protection scope of the present invention.

[0039] In order to better illustrate the present invention, the present invention will be described in detail below in conjunction with the accompanying drawings.

[0040] 1. Genome extraction and sequencing of Streptomyces grisea

[0041] Inoculate the spore suspension of Streptomyces grisea into LB liquid medium, culture at 37°C and 150rpm for 3 days, then collect 2.0mL of the bacterial liquid, centrifuge at 12000rpm for 2min, discard the supernatant, and collect the bacterial precipitate. Add 180 μL lysozyme (20 mg / mL) and 20 μl EDTA solutio...

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Abstract

The invention discloses streptomyces griseorubens pectinesterase as well as a coding gene and an application thereof. The pectinesterase gene is cloned from streptomyces griseorubens; a nucleotide sequence of the gene is shown as SEQ ID NO.1 or is shown as SEQ ID NO.2 after codon optimization; an amino acid sequence of the coded pectinesterase is shown as SEQ ID NO.3; and the invention also discloses applications of the pectinesterase and the coding gene thereof in degrading pectin. According to the invention, by adding a soluble label to target protein and through peripheral space expression of the target protein, the bio-activity of the protein is guaranteed, and meanwhile, subsequent protein purification is facilitated. The pectinesterase is alkaline and high-temperature resistant, and the enzyme (pectinesterase) has a quite strong resisting capacity to a plurality of metal ions and enzymatic activity inhibitors; therefore, the enzyme has great significance in practical production and application.

Description

technical field [0001] The invention relates to the technical fields of genetic engineering and modern enzyme engineering, and relates to a pectin esterase of Streptomyces grisea and its encoding gene and its application, in particular to a pectin esterase and its encoding gene sequence, including the encoding gene sequence The recombinant expression vector, the transgenic recombinant bacteria and the application of the pectin esterase. Background technique [0002] Pectinase is widely present in fruits and various microorganisms, and is a general term for a class of complex enzymes composed of many single enzymes that can act on pectin. Pectinase has main application significance in the fields of ramie degumming, wood preservation and alcohol fermentation, and has been widely used in the food industry. [0003] Pectin esterase (EC 3.1.1.11) is an enzyme that catalyzes and hydrolyzes pectin to produce pectic acid and methanol. Pectinase has high specificity to polygalactur...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/18C12N15/55C12N15/70C12N1/21C12R1/19
CPCC12N9/18C12N15/70C12N2800/101C12Y301/01011
Inventor 周培支月娥冯海玮孙玉静毛亮罗艳青聂文翰
Owner SHANGHAI JIAO TONG UNIV
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