High-level secretion of a Rhizopus niveus aspartic proteinase in Saccharomyces cerevisiae.
The gene encoding an extracellular Rhizopus niveus aspartic proteinase I (RNAP-I) was introduced into Saccharomyces cerevisiae. The yeast cell carrying a plasmid containing the intact RNAP-I gene under the control of the glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene promoter of S. cerevisiae did not synthesize RNAP-I at all. On the other hand, when the intron of the RNAP-I gene had been removed from the gene in the plasmid, the cell secreted RNAP-I with high efficiency. Processing of the pro-sequence occurred at the same region of the pro-enzyme during cultivation as observed in the culture of R. niveus. Moreover, the promoter and the terminator of the original RNAP-I gene were found to be weakly functional in the yeast cell with respect to expression of the intronless RNAP-I gene, although the initiation and termination sites were heterogeneous. The effects of vector-types on the extracellular production of RNAP-I were also investigated.
Horiuchi
H
H
Department of Agricultural Chemistry, University of Tokyo, Japan.
Ashikari
T
T
Amachi
T
T
Yoshizumi
H
H
Takagi
M
M
Yano
K
K
D016282:Aspartic Acid Endopeptidases / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D005110:Extracellular Space / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D005658:Fungi / Q000201:enzymology* / Q000235:genetics; D015870:Gene Expression; D005796:Genes / Q000502:physiology; D007438:Introns / Q000502:physiology; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011506:Proteins / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D014162:Transfection
D005656:Fungal Proteins; D011506:Proteins; D016282:Aspartic Acid Endopeptidases; C100354:aspartic proteinase-I