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KMID : 0613820020120040477
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Journal of Life Science
2002 Volume.12 No. 4 p.477 ~ p.482
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Expression of Biologically Active Insect-Derived Antibacterial Peptide, Defensin, in Yeast
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Kang Dae-Ock
Ahn Soon-Cheol
Kim Min-Soo
Ahn Jong-Seog
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Abstract
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As a biological model system for the production of an active antibacterial peptide, we have attempted the expression and secretion of insect defensin in Saccharomyces cerevisiae. Nucleotide sequences encoding mature defensin composed of 40 amino acids were fused in frame with promoter and signal sequence of Saccharomyces diastaticus glucoamylase, and mating factor α1[MFα1] prosequence. The host strain, S. cerevisiae 2805 was transformed with the resulting plasmid, pSMF11. The secretion of functional defensin was confirmed by growth inhibition zone assay using Micrococcus luteus as a test organism. Insect defensin was secreted to the culture supernatant in biologically active form by glucoamylase signal sequence and mating factor α1 prosequence. Most of antibacterial activity was detected in the culture supernatant. Defensin was also active against Staphylococcus aureus and Listeria monocytogenes.
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KEYWORD
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Defensin, glucoamylase signal sequence, mating factor α1 prosequence, secretion, Saccharomyces cerevisiae
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