Kluyveromyces as a host for heterologous gene expression: expression and secretion of prochymosin.
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.
Royal Gist-brocades, Research and Development, Ma Delft, The Netherlands.
J A
JA
van den Berg
K J
KJ
van der Laken
A J
AJ
van Ooyen
T C
TC
Renniers
K
K
Rietveld
A
A
Schaap
A J
AJ
Brake
R J
RJ
Bishop
K
K
Schultz
D
D
Moyer
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D012085:Chymosin / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D004792:Enzyme Precursors / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D007716:Kluyveromyces / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D010957:Plasmids; D011499:Protein Processing, Post-Translational; D021382:Protein Sorting Signals / Q000235:genetics / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D004718:Saccharomycetales / Q000235:genetics*; D013045:Species Specificity
D004792:Enzyme Precursors; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; C032443:prorennin; D012085:Chymosin