Expression, Purification and Antibacterial Activity of NK-Lysin Mature Peptides from the Channel Catfish (Ictalurus punctatus)

被引：21

作者：

Cai, Shurui ^{[1
]}

Wang, Jun ^{[2
]}

Wang, Kaiyu ^{[2
]}

Chen, Defang ^{[3
]}

Dong, Xiaowei ^{[1
]}

Liu, Tao ^{[2
]}

Zeng, Yukun ^{[2
]}

Wang, Xingli ^{[2
]}

Wu, Dongmei ^{[1
]}

机构：

[1] Sichuan Agr Univ, Coll Vet Med, Wenjiang Dist Huimin Rd 211, Chengdu 611130, Sichuan, Peoples R China

[2] Sichuan Agr Univ, Dept Basic Vet Med, Key Lab Anim Dis & Human Hlth Sichuan Prov, Wenjiang Dist Huimin Rd 211, Chengdu 611130, Sichuan, Peoples R China

[3] Sichuan Agr Univ, Dept Aquaculture, Wenjiang Dist Huimin Rd 211, Chengdu 611130, Sichuan, Peoples R China

来源：

APPLIED SCIENCES-BASEL | 2016年 / 6卷 / 09期

关键词：

NK-lysins; expression; purification; channel catfish; antibacterial activity; HOST-DEFENSE PEPTIDES; ANTIMICROBIAL PEPTIDE; ESCHERICHIA-COLI; FUSION EXPRESSION; INNATE IMMUNITY; T-CELLS; GRANULYSIN; BACTERIAL; GENE; SECRETIONS;

D O I：

10.3390/app6090240

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

Antimicrobial peptides (AMPs) are small peptides and play important roles in host innate immune response against microbial invasion. Aquatic animals secrete different kinds of antimicrobial peptides which have antimicrobial activity towards microorganisms. NK-lysins, mature peptides produced by cytotoxic T lymphocytes and natural killer cells, are comprised of 74-78 amino acid residues, demonstrating broad-spectrum antimicrobial activity against bacteria, fungi, protozoa, and parasites. In this study, three distinct NK-lysin mature peptide (mNKLs), transcripts (76 amino acid residues) cloned from the channel catfish (Ictalurus punctatus) head kidney were ligated into plasmid vector pET-32a(+) to express the mNKLs fusion protein. The fusion protein was successfully expressed in E. coli Rosetta (DE3) under optimized conditions. After purification by affinity column chromatography, the fusion protein was successfully cleaved by enterokinase and released the peptide mNKLs. Tricine-SDS-PAGE results showed that mNKLs (approximately 8.6 kDa) were successfully expressed. The purified peptide mNKLs exhibited antibacterial activity against Staphylococcus aureus and E. coli.

引用

页数：11

共 57 条

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