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Ruminant rumen-specific lysozyme lyz1 and its application

A technology of ruminants and lysozyme, which is applied in the field of bioengineering, can solve the problems that lysozyme has not yet been discovered and its safety is unknown, and achieves the effect of easy preparation, separation and purification

Active Publication Date: 2022-07-15
NORTHWEST A & F UNIV
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

To date, a variety of lysozymes have been identified from ruminants, but there are still many species of lysozymes that have not yet been discovered
And for the lysozyme that has been prepared (for example, Fu Shixin et al. reported "secretion, expression and activity analysis of milk lysozyme in Pichia pastoris"), its safety as a feed additive for directly feeding animals and as a substitute for antibiotics sex is unknown

Method used

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  • Ruminant rumen-specific lysozyme lyz1 and its application

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Embodiment Construction

[0025] The present invention will be described in further detail below with reference to the accompanying drawings and embodiments, so as to facilitate those skilled in the art to understand the present invention, but the protection scope of the present invention is not limited thereto.

[0026] (1) Secretion and expression method of ruminant-specific lysozyme LYZ1 in Pichia pastoris

[0027] 1. Construction of Pichia pastoris GS115 recombinant strain

[0028] 1.1 The present invention uses the results of comparative genomes of ruminants to scan new genes evolved in ruminants, and finds a new ruminant-specific lysozyme, which is specifically highly expressed in the rumen. According to the amino acid sequence of lysozyme LYZ1 obtained by comparing ruminant genomes and according to the codon preference of Pichia pastoris, the recombinant goat (Capra hircus) lysozyme LYZ1 gene sequence (synthesized by Wuhan Jinkarui Bioengineering Co., Ltd.) with the signal peptide removed was ob...

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Abstract

The invention discloses a ruminant rumen-specific lysozyme LYZ1 and its application. The invention utilizes the comparative genome results of ruminants to scan the new genes evolved by ruminants, and finds a ruminant-specific lysozyme, which is highly expressed in rumen tissue. The present invention utilizes pPIC9K vector and ruminant rumen-specific lysozyme LYZ1 sequence to construct methanol-inducible Pichia pastoris expression strain, so that ruminant animal rumen-specific lysozyme LYZ1 can be efficiently secreted and expressed in Pichia pastoris. The representative lysozyme LYZ1 has the effect of inhibiting Staphylococcus aureus and can be used as a potential and safe alternative to antibiotics.

Description

technical field [0001] The invention belongs to the field of bioengineering, and in particular relates to a method and application for the secretion and expression of ruminant rumen-specific lysozyme 1 (lysozyme-1, LYZ1) in Pichia pastoris. Background technique [0002] As a natural antibacterial agent, lysozyme widely exists in various tissues and organs of humans and mammals. It has various pharmacological effects against bacteria, fungi and tumors. It has good biocompatibility and no irritation to tissues. , non-toxic characteristics, has been made as an enzyme preparation with excellent properties, killing pathogenic microorganisms without destroying the body. Lysozyme is divided into 6 categories according to different molecular structures and sources: plant lysozyme, microbial lysozyme, phage lysozyme, type C (egg white lysozyme, stomach lysozyme), type G (goose type lysozyme) and type I (no lysozyme) Vertebrate lysozyme), of which C-type, G-type and phage lysozyme ar...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/36C12N15/56C12N15/81A23K20/189A61K38/47A61P31/04
CPCC12N9/2462C12Y302/01017C12N15/815A23K20/189A61K38/47A61P31/04
Inventor 姜雨潘香羽陈贤情王文
Owner NORTHWEST A & F UNIV
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