title: Characterization of a novel manganese dependent endoglucanase belongs in GH family 5 from Phanerochaete chrysosporium.
abstract: The cDNA encoding a putative glycoside hydrolase family 5, which has been predicted to be an endoglucanase (PcEg5A), was cloned from Phanerochaete chrysosporium and expressed in Pichia pastoris. PcEg5A contains a carbohydrate-binding domain and two important amino acids, E209 and E319, playing as proton donor and nucleophile in substrate catalytic domain. SDS-PAGE analysis indicated that the recombinant endoglucanase 5A (rPcEg5A) has a molecular size of 43 kDa which corresponds with the theoretical calculation. Optimum pH and temperature were found to be 4.5-6.0, and 50°C-60°C, respectively. Moreover, rPcEg5A exhibited maximal activity in the pH range of 3.0-8.0, whereas over 50% of activity still remained at 20°C and 80°C. rPcEg5A was stable at 60°C for 12 h incubation, indicating that rPcEg5A is a thermostable enzyme. Manganese ion enhanced the enzyme activity by 77%, indicating that rPcEg5A is a metal dependent enzyme. The addition of rPcEg5A to cellobiase (cellobiohydrolase and β-glucosidase) resulted in a 53% increasing saccharification of NaOH-pretreated barley straw, whereas the glucose release was 47% higher than that cellobiase treatment alone. Our study suggested that rPcEg5A is an enzyme with great potential for biomass saccharification.
authors.affiliation: Division of Biotechnology, College of Environmental and Bioresource Sciences, Chonbuk National University, Iksan, Jeonbuk 570-752, Republic of Korea; Institute of Biotechnology, Hue University, Hue 530000, Viet Nam.
authors.forename: Nguyen Duc
authors.initials: ND
authors.lastname: Huy
authors.affiliation: Division of Biotechnology, College of Environmental and Bioresource Sciences, Chonbuk National University, Iksan, Jeonbuk 570-752, Republic of Korea.
authors.forename: Cu Le
authors.initials: CL
authors.lastname: Nguyen
authors.affiliation: Division of Biotechnology, College of Environmental and Bioresource Sciences, Chonbuk National University, Iksan, Jeonbuk 570-752, Republic of Korea.
authors.forename: Han-Sung
authors.initials: HS
authors.lastname: Park
authors.affiliation: College of Sciences, Hue University, Hue 530000, Viet Nam.
authors.forename: Nguyen Hoang
authors.initials: NH
authors.lastname: Loc
authors.affiliation: Institute of Molecular Biology and Genetics, College of Natural Sciences, Chonbuk National University, Jeonju, Jeonbuk 561-756, Republic of Korea.
authors.forename: Myoung-Suk
authors.initials: MS
authors.lastname: Choi
authors.affiliation: Institute of Molecular Biology and Genetics, College of Natural Sciences, Chonbuk National University, Jeonju, Jeonbuk 561-756, Republic of Korea.
authors.forename: Dae-Hyuk
authors.initials: DH
authors.lastname: Kim
authors.affiliation: Applied Microbiology Research Center, Bio-Materials Research Institute, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup, Jeonbuk 580-185, Republic of Korea.
authors.forename: Jeong-Woo
authors.initials: JW
authors.lastname: Seo
authors.affiliation: Division of Biotechnology, College of Environmental and Bioresource Sciences, Chonbuk National University, Iksan, Jeonbuk 570-752, Republic of Korea. Electronic address: smpark@chonbuk.ac.kr.
authors.forename: Seung-Moon
authors.initials: SM
authors.lastname: Park
mesh_terms: D018533:Biomass; D020134:Catalytic Domain; D002480:Cellulase / Q000737:chemistry / Q000145:classification* / Q000235:genetics / Q000378:metabolism*; D018076:DNA, Complementary / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D005947:Glucose / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008345:Manganese / Q000378:metabolism*; D008970:Molecular Weight; D020075:Phanerochaete / Q000201:enzymology* / Q000235:genetics; D010843:Pichia / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D013696:Temperature; D001617:beta-Glucosidase / Q000378:metabolism
chemical_list: D018076:DNA, Complementary; D011994:Recombinant Proteins; D008345:Manganese; D001617:beta-Glucosidase; D002480:Cellulase; D005947:Glucose
keywords: Barley straw; Endoglucanase; Manganese dependent enzyme; Phanerochaete chrysosporium; Saccharification