Expression of two Trichoderma reesei xylanases in the fission yeast Schizosaccharomyces pombe

Two xylanase genes (xyn1 and xyn2) were amplified by the polymerase chain reaction (PCR) technique from first-strand cDNA prepared from mRNA of Trichoderma reesei QM9414. The genes were located under the human cytomegalovirus gene promoter (CMVp) on copy-number-controlled plasmids (pTLxyn1 and pTLxyn2). When both plasmids were introduced into Schizosaccharomyces pombe, functional xylanases (XYN I and XYN II) were secreted by the recombinant yeasts. The secreted XYN I protein had a molecular mass of 21 kDa whereas XYN II was produced as two molecular forms with sizes of 21 and 28 kDa, the former being not glycosylated and the latter N-glycosylated. XYN I was secreted in the culture medium at a level of about 25 microg/ml and XYN II at about 170 microg/ml. The recombinant xylanases had the same characteristics with respect to the effects of temperature and pH on the enzyme activity as the native ones.