T. Hatano et al., EXPRESSION OF THE CARBOXYMETHYLCELLULASE GENE, CMC1, FROM CRYPTOCOCCUS-FLAVUS IN SACCHAROMYCES-CEREVISIAE, Journal of fermentation and bioengineering, 77(3), 1994, pp. 235-238
A cDNA copy of the carboxymethylcellulase (CMCase 1) gene, CMC1, of th
e yeast Cryptococcus flavus was expressed in Saccharomyces cerevisiae
under the control of the glyceraldehyde-3-phosphate dehydrogenase gene
(GAP) promoter derived from S. cerevisiae. In glucose medium, extrace
llular production of a large amount of CMCase 1 was achieved by S. cer
evisiae cells transformed with YEp-plasmid carrying CMC1. Two major pr
oteins (36 kDa and 34 kDa) secreted in the medium both had CMCase acti
vity and immuno-responsibility to anti-CMCase 1-antibody. From an anal
ysis of amino acid sequences, it was found that the 36 kDa protein is
a pro-CMCase 1 having an N-terminal residue, Ala, the 19th amino acid
of the peptide deduced from the nucleotide sequence of CMC1 on the cDN
A, and that the 34 kDa protein is a mature CMCase 1. Alteration of the
cell morphology of S. cerevisiae was induced by transformation with Y
Ep-plasmid carrying CMC1.