Kluyveromyces as a host for heterologous gene expression: expression and secretion of prochymosin

• Published in: Bio/technology (New York, N.Y. 1983) Vol. 8; no. 2; pp. 135 - 139
• Main Authors: Berg, J.A. van den (Royal Gist-brocades, The Netherlands), Laken, K.J. van der, Van Ooyen, A.J.J, Renniers, T.C.H.M, Rietveld, K, Schaap, A, Brake, A.J, Bishop, R.J, Schultz, K, Moyer, D
• Format: Journal Article
• Language: English
• Published: New York, NY Nature Publications 01-02-1990
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Biological and medical sciences; BIOSINTESIS; BIOSYNTHESE; BIOSYNTHESIS; Biotechnology; Cattle; CHYMOSIN; Chymosin - biosynthesis; Chymosin - genetics; Chymosin - secretion; CHYMOSINE; Cloning, Molecular; Enzyme Precursors - biosynthesis; Enzyme Precursors - genetics; Enzyme Precursors - secretion; Fundamental and applied biological sciences. Psychology; GENE; GENES; Genetic engineering; Genetic technics; KLUYVEROMYCES; Kluyveromyces - genetics; Kluyveromyces - metabolism; KLUYVEROMYCES LACTIS; Methods. Procedures. Technologies; Molecular Sequence Data; Plasmids; Protein Processing, Post-Translational; Protein Sorting Signals - genetics; Protein Sorting Signals - physiology; PROTEINAS; PROTEINE; PROTEINS; QUIMOSINA; Recombinant Fusion Proteins - biosynthesis; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - secretion; Saccharomyces cerevisiae - genetics; Saccharomycetales - genetics; SECRECION; SECRETION; Species Specificity; Fungi; Kluyveromyces marxianus var. lactis; Enzyme; Secretion; Chymosin; Production; Immunoblotting assay; Ascomycetes; Genetic engineering; Gene expression; Precursor; Thallophyta
• ISSN: 0733-222X; 2331-3684
• DOI: 10.1038/nbt0290-135

We have developed the yeast Kluyveromyces lactis as a host organism for the production of the milk-clotting enzyme chymosin. In contrast to Saccharomyces cerevisiae, we found that this yeast is capable of the synthesis and secretion of fully active prochymosin. Various signal sequences could be used to efficiently direct the secretion of prochymosin in Kluyveromyces, but not in S. cerevisiae. We conclude that the efficient synthetic and secretory capacity of this heterologous protein is a property of the yeast Kluyveromyces. These results have led to the development of a large scale production process for chymosin.