Overproduction of recombinant Trichoderma reesei cellulases by Aspergillus oryzae and their enzymatic properties.
We have established an expression system of Trichoderma reesei cellulase genes using Aspergillus oryzae as a host. In this system, the expression of T. reesei cellulase genes were regulated under the control of A. oryzae Taka-amylase promoter and the cellulase genes were highly expressed when maltose was used as a main carbon source for inducer. The production of recombinant cellulases by A. oryzae transformants reached a maximum after 3-4 days of cultivation. In some cases, proteolysis of recombinant cellulases was observed in the late stage of cultivation. The recombinant cellulases were purified and characterized. The apparent molecular weights of recombinant cellulases were more or less larger than those of native enzymes. The optimal temperatures and pHs of recombinant cellulases were 50-70 degrees C and 4-5, respectively. Among the recombinant cellulases, endoglucanase I showed broad substrate specificities and high activity when compared with the other cellulases investigated here.
Takashima
S
S
Department of Biotechnology, Graduate School of Agricultural and Life Sciences, University of Tokyo, Japan.
Iikura
H
H
Nakamura
A
A
Hidaka
M
M
Masaki
H
H
Uozumi
T
T
D001236:Aspergillus oryzae / Q000235:genetics*; D015139:Blotting, Southern; D002480:Cellulase / Q000096:biosynthesis* / Q000302:isolation & purification / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D015966:Gene Expression Regulation, Fungal; D005822:Genetic Vectors; D006026:Glycoside Hydrolases; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D013379:Substrate Specificity; D014242:Trichoderma / Q000201:enzymology* / Q000235:genetics; D015003:Yeasts / Q000201:enzymology
D011994:Recombinant Proteins; D006026:Glycoside Hydrolases; D002480:Cellulase; C100181:pullulanase