Eukaryotic expression and antimicrobial spectrum determination of the peptide tachyplesin II.
Ta0-a, the gene encoding the mature antimicrobial peptide tachyplesin II, was engineered to optimize the coding sequence according to codon usage bias in yeast. Ta0-a was efficiently expressed in the methylotrophic yeast Pichia pastoris strain SMD1168. The recombinant peptide Ta0 reached 150mg/L after methanol induction for 6 d. Ta0 was rapidly purified to homogeneity by a single step of size-exclusion chromatography. The minimal lethal concentrations of Ta0 to the Escherichia coli strain K12 was 30 microg/mL. Ta0 exhibited a wide range of antimicrobial activity: the growth of 26 bacterial and fungal strains, including some typical food/feed spoilage microorganisms, was all substantially inhibited. This result indicates the potential practical application of the recombinant peptide in various industrial products.
Xu
Feng
F
Microbial Engineering Department, Feed Research Institute, Chinese Academy of Agricultural Sciences, No. 12, Zhongguancun South Street, Haidan District, Beijing 100081, China; College of Life Science, Yangtze University, Jingsha, Hubei 434100, China.
Meng
Kun
K
Wang
Ya-Ru
YR
Luo
Hui-Ying
HY
Yang
Pei-Long
PL
Wu
Ning-Feng
NF
Fan
Yun-Liu
YL
Yao
Bin
B
D000595:Amino Acid Sequence; D023181:Antimicrobial Cationic Peptides / Q000096:biosynthesis* / Q000494:pharmacology*; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000096:biosynthesis* / Q000494:pharmacology*; D004220:Disulfides / Q000032:analysis; D004926:Escherichia coli / Q000187:drug effects / Q000378:metabolism; D005658:Fungi / Q000187:drug effects; D008826:Microbial Sensitivity Tests; D010456:Peptides, Cyclic / Q000096:biosynthesis* / Q000494:pharmacology*; D010843:Pichia / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000494:pharmacology; D012475:Salmonella / Q000187:drug effects; D012762:Shigella dysenteriae / Q000187:drug effects; D013211:Staphylococcus aureus / Q000187:drug effects; D015003:Yeasts / Q000187:drug effects
D023181:Antimicrobial Cationic Peptides; D004268:DNA-Binding Proteins; D004220:Disulfides; D010456:Peptides, Cyclic; D011994:Recombinant Proteins; C057636:tachyplesin peptide, Tachypleus tridentatus