Characterization of endo-β-1,4-glucanase from a novel strain of Penicillium pinophilum KMJ601

• Published in: Applied microbiology and biotechnology 2010, Vol.85 (4), p.1005-1014
• Main Authors: Jeya, Marimuthu, Joo, Ah-Reum, Lee, Kyoung-Mi, Sim, Won-Il, Oh, Deok-Kun, Kim, Yeong-Suk, Kim, In-Won, Lee, Jung-Kul
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Base Sequence; Biological and medical sciences; Biomedical and Life Sciences; Biotechnologically Relevant Enzymes and Proteins; Biotechnology; carbon; Carbon - metabolism; Cellulase - biosynthesis; Cellulase - genetics; Cellulase - isolation & purification; Cloning, Molecular; corn straw; DNA, Fungal; electrophoresis; Electrophoresis, Polyacrylamide Gel; Fermentation; Fundamental and applied biological sciences. Psychology; Fungal Proteins - genetics; genes; Genes, Fungal; Hydrogen-Ion Concentration; internal transcribed spacers; Life Sciences; Microbial Genetics and Genomics; Microbiology; microorganisms; Molecular Sequence Data; nitrogen; Nitrogen - metabolism; Penicillium; Penicillium - enzymology; Penicillium - isolation & purification; Polymerase Chain Reaction; Proteomics; ribosomal DNA; rice straw; Temperature
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-009-2070-0

A novel endo-β-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°C and 5.0, respectively. The increased production of EG by P. pinophilum in culture at 28°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.