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KMID : 0545119930030020065
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Journal of Microbiology and Biotechnology
1993 Volume.3 No. 2 p.65 ~ p.72
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Heterologous Gene Expression and Secretion of the Anticoagulant Hirudin in a Methylotrophic Yeast Hansenula polymorpha
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SOHN, JUNG HOON
BEBUROV, MICHAEL YU/CHOI, DUI SUNG/RHEE, SANG KI
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Abstract
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A heterologous gene expression and secretion system using a methylotrophic yeast, Hansenula polymorpha was developed for the production of anticoagulant hirudin. Hirudin gene was expressed under the control of a strong and inducible methanol oxidase (MOX or AOX) promoter. The mating factor α pre-pro leader sequence of Saccharomyces cerevisiae was employed for hirudin to be secreted into the extracellular medium. Hirudin expression cassette was introduced into three strains of H. polymorpha, A16, HPB1 and DL1 which have different genetic backgrounds. This expression cassette was stably integrated into the host chromosomal DNA. Biologically active and mature hirudin was efficiently expressed and secreted into the extracellular medium. About 19㎎/L of hirudin was found in the culture supernatant in the case of a two-copy integrant of the strain HPB1 under suboptimal culture conditions.
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KEYWORD
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