Characterization of endo-beta-1,4-glucanase from a novel strain of Penicillium pinophilum KMJ601.
A novel endo-beta-1,4-glucanase (EG)-producing strain was isolated and identified as Penicillium pinophilum KMJ601 based on its morphology and internal transcribed spacer (ITS) rDNA gene sequence. When rice straw and corn steep powder were used as carbon and nitrogen sources, respectively, the maximal EG activity of 5.0 U mg protein(-1), one of the highest levels among EG-producing microorganisms, was observed. The optimum temperature and pH for EG production were 28 degrees C and 5.0, respectively. The increased production of EG by P. pinophilum in culture at 28 degrees C was confirmed by two-dimensional electrophoresis followed by MS/MS sequencing of the partial peptide. A partial EG gene (eng5) was amplified by degenerate polymerase chain reaction (PCR) based on the peptide sequence. A full-length eng5 was cloned by genome-walking PCR, and P. pinophilum EG was identified as a member of glycoside hydrolase family 5. The present results should contribute to improved industrial production of EG by P. pinophilum KMJ601.
Jeya
Marimuthu
M
Department of Chemical Engineering, Konkuk University, Seoul, South Korea.
Joo
Ah-Reum
AR
Lee
Kyoung-Mi
KM
Sim
Won-Il
WI
Oh
Deok-Kun
DK
Kim
Yeong-Suk
YS
Kim
In-Won
IW
Lee
Jung-Kul
JK
D000595:Amino Acid Sequence; D001483:Base Sequence; D002244:Carbon / Q000378:metabolism; D002480:Cellulase / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D003001:Cloning, Molecular; D004271:DNA, Fungal; D004591:Electrophoresis, Polyacrylamide Gel; D005285:Fermentation; D005656:Fungal Proteins / Q000235:genetics; D005800:Genes, Fungal; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D009584:Nitrogen / Q000378:metabolism; D010407:Penicillium / Q000201:enzymology* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction; D040901:Proteomics; D013696:Temperature
D004271:DNA, Fungal; D005656:Fungal Proteins; D002244:Carbon; D002480:Cellulase; D009584:Nitrogen