Effect of culture temperature on the heterologous expression of Pleurotus eryngii versatile peroxidase in Aspergillus hosts.
Production of recombinant versatile peroxidase in Aspergillus hosts was optimized through the modification of temperature during bioreactor cultivations. To further this purpose, the cDNA encoding a versatile peroxidase of Pleurotus eryngii was expressed under control of the alcohol dehydrogenase (alcA) promoter of Aspergillus nidulans. A dependence of recombinant peroxidase production on cultivation temperature was found. Lowering the culture temperature from 28 to 19 degrees C enhanced the level of active peroxidase 5.8-fold and reduced the effective proteolytic activity twofold. Thus, a maximum peroxidase activity of 466 U L(-1) was reached. The same optimization scheme was applied to a recombinant Aspergillus niger that bore the alcohol dehydrogenase regulator (alcR), enabling transformation with the peroxidase cDNA under the same alcA promoter. However, with this strain, the peroxidase activity was not improved, while the effective proteolytic activity was increased between 3- and 11-fold compared to that obtained with A. nidulans.
Eibes
G M
GM
Department of Chemical Engineering, Institute of Technology, Santiago de Compostela, Spain. geibes@usc.es
Lú-Chau
T A
TA
Ruiz-Dueñas
F J
FJ
Feijoo
G
G
Martínez
M J
MJ
Martínez
A T
AT
Lema
J M
JM
D000426:Alcohol Dehydrogenase / Q000235:genetics / Q000378:metabolism; D001230:Aspergillus / Q000378:metabolism*; D001233:Aspergillus nidulans / Q000378:metabolism; D001234:Aspergillus niger / Q000201:enzymology; D019149:Bioreactors; D001709:Biotechnology / Q000379:methods; D018076:DNA, Complementary / Q000378:metabolism; D005285:Fermentation; D007218:Industrial Microbiology / Q000379:methods; D008828:Microbiological Techniques; D009195:Peroxidase / Q000737:chemistry*; D020076:Pleurotus / Q000201:enzymology*; D011994:Recombinant Proteins / Q000737:chemistry; D013696:Temperature; D013997:Time Factors
D018076:DNA, Complementary; D011994:Recombinant Proteins; D000426:Alcohol Dehydrogenase; D009195:Peroxidase