杂合抗菌肽Mel-MytB在毕赤酵母中的表达及其抗菌活性测定

doi:10.16431/j.cnki.1671-7236.2018.11.032

Abstract

Abstract:

In order to abtain a high activity antimicrobial peptide,an expression vector was constructed with a tandem of Melittin and Mytilin-B.According to the partiality codon of Pichia pastoris,the Melittin(3-14)-Mytilin-B(13-27) hybrid antimicrobial peptide,Mel-MytB (MEM) gene was synthesized by SOE method,and cloned into pPICZa-A to construct the recombinant expression vector pPICZa-A-MEM.pPICZα-A-MEM was linearized by Sac Ⅰ and electroporated into Pichia pastoris X-33.The results showed that under the control of the promoter alcohol oxidase (AOX),an approximately 3.0 ku Mel-MytB protein was expressed,which had thermal stability and acid stability.The antibacterial activity was basically unchanged after boiling for 40 min and pH 2.0-10.0.Antibacterial assays demonstrated that Mel-MytB had broad-spectrum antibacterial activity.The MIC to Escherichia coli ATCC25922,Staphylococcus aureus ATCC25923,Staphylococcus aureus ATCC6538,Salmonella enterica ATCC13076,Salmonella Typhimurium ATCC14028,Vibrio parahaemolyticus ATCC17802,Vibrio vulnificus ATCC27562 and Bacillus subtilis ATCC6633 were 5.1,2.2,2.0,4.6,7.6,9.4,13.2 and 59.4 μg/mL,respectively.Therefore,the recombinant antimicrobial peptide Mel-MytB had a good application prospect in disease prevention and animal feed additives.

Key words: hybrid antimicrobial peptide; Melittin; Mytilin-B; Pichia pastoris; antibacterial activity

CLC Number:

S859.79⁺7

ZHANG Honggang, WU Jianliang, LI Li. Expression of Hybrid Antimicrobial Peptide Mel-MytB in Pichia pastoris and Detection of Its Antibacterial Activity[J]. , 2018, 45(11): 3246-3252.

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Expression of Hybrid Antimicrobial Peptide Mel-MytB in Pichia pastoris and Detection of Its Antibacterial Activity

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