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Low-temperature alkaline phosphatidase A1 and coding gene thereof

A phospholipase and alkaline technology, applied in genetic engineering, plant genetic improvement, hydrolytic enzymes, etc., to achieve the effect of simplifying the production process

Inactive Publication Date: 2011-03-30
THE INST OF MICROBIOLOGY XINJIANG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the research on Serratia that produces both low-temperature phospholipase and lipase has not been reported at home and abroad.

Method used

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  • Low-temperature alkaline phosphatidase A1 and coding gene thereof
  • Low-temperature alkaline phosphatidase A1 and coding gene thereof
  • Low-temperature alkaline phosphatidase A1 and coding gene thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Embodiment 1: low temperature alkaline phospholipase A 1 Screening of producing bacteria

[0047] 1. Isolation and screening of strains

[0048] Collect Xinjiang Tianshan No. 1 glacier and permafrost, and apply it with 0.1% (NH 4 ) 2 SO 4 , 0.1%K 2 HPO 4 , 1% KCl, 0.05% MgSO 4 ·7H 2 O, 0.001% FeSO 4 ·7H 2 O, olive oil and polyvinyl alcohol emulsion mixed in a ratio of 1: 3, on the fat assimilation medium of 2% agar powder, cultivated at 20°C for 24 hours and screened 3 lipase-producing bacterial strains. Bacterial colonies were selected and spread on egg yolk+TYSPN solid agar medium (4% glucose, 1% peptone, 5% yeast extract, sterile egg yolk liquid, 2% agar powder, pH7.0). The strain xjFl formed a yellow discoloration circle on the oil assimilation plate, and formed an opaque and cloudy halo on the egg yolk + TYSPN solid agar plate. Insert strain xjF1 into phospholipase production medium (5.4% peptone, 2.6% yeast extract, 1.512% NaH 2 PO 4 12H 2 O, 0.3%K ...

Embodiment 2

Embodiment 3

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Abstract

This invention discloses a cold-resistant Serratia fonticola xjFl CGMCC No.1971. Through the research of the strain characteristics, enzyme production conditions and the nature of enzyme, a new phospholipase A1 gene is found to consists of two open reading frames plA and plS, and the plA gene is certified to coding the phospholipase A1, the plS gene is certified to coding the phospholipase A1 auxiliary protein. This invention relates to a recombinant plasmid and a recombinant strain containing the gene, the gene expression product phospholipase A1 in the shows activity under low-temperature conditions, which can be used in industries such as oil refining process to simplify the production process, save the energy consumption, improve the product quality, protect the environment, reduce the production costs, and have important practical significance.

Description

technical field [0001] The invention belongs to the fields of enzyme genetic engineering and enzyme engineering. Specifically, the present invention relates to a kind of low-temperature alkaline phospholipase A of coding resistance Serratia cold springs 1 The gene sequence of the enzyme, the recombinant plasmid containing the enzyme gene, and the recombinant strains and derivatives expressing the corresponding enzyme. Background technique [0002] Phospholipases are enzymes that hydrolyze the ester bonds of phospholipids. Widely present in eukaryotes and prokaryotes, it affects the catabolism of phospholipids, the formation and reorganization of biofilms, and phospholipases are also involved in the cascade of signals. Several types of phospholipases are known with different specificities depending on the position of the bond that the phospholipase molecule attacks, of which phospholipase A 1 (Phospholipase A 1 , 3.1.1.32, referred to as PLA 1 ) catalyzes the hydrolysis ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N9/16C12N15/55C12N1/21C12N1/19C12R1/425C12R1/19C12R1/885
Inventor 付建红姚斌石玉瑚孟昆欧阳平凯袁铁铮
Owner THE INST OF MICROBIOLOGY XINJIANG ACADEMY OF AGRI SCI
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