title: Characterization of a novel endo-β-1,4-glucanase from Armillaria gemina and its application in biomass hydrolysis.
abstract: A novel endo-β-1,4-glucanase (EG)-producing strain was isolated and identified as Armillaria gemina KJS114 based on its morphology and internal transcribed spacer rDNA gene sequence. A. gemina EG (AgEG) was purified using a single-step purification by gel filtration. The relative molecular mass of AgEG by sodium dodecyl sulfate polyacrylamide gel electrophoresis was 65 kDa and by size exclusion chromatography was 66 kDa, indicating that the enzyme is a monomer in solution. The pH and temperature optima for hydrolysis were 5.0 and 60 °C, respectively. Purified AgEG had the highest catalytic efficiency with carboxymethylcellulose (k(cat)/K(m) = 3,590 mg mL⁻¹ s⁻¹) unlike that reported for any fungal EG, highlighting the significance of the current study. The amino acid sequence of AgEG showed homology with hydrolases from the glycoside hydrolase family 61. The addition of AgEG to a Populus nigra hydrolysate reaction containing a commercial cellulase mixture (Celluclast 1.5L and Novozyme 188) showed a stimulatory effect on reducing sugar production. AgEG is a good candidate for applications that convert lignocellulosic biomass to biofuels and chemicals.
authors.affiliation: Department of Chemical Engineering, Konkuk University, 1 Hwayang-Dong, Gwangjin-Gu, Seoul, 143-701, Republic of Korea.
authors.forename: Sujit Sadashiv
authors.initials: SS
authors.lastname: Jagtap
authors.forename: Saurabh Sudha
authors.initials: SS
authors.lastname: Dhiman
authors.forename: Tae-Su
authors.initials: TS
authors.lastname: Kim
authors.forename: In-Won
authors.initials: IW
authors.lastname: Kim
authors.forename: Jung-Kul
authors.initials: JK
authors.lastname: Lee
mesh_terms: D055392:Armillaria / Q000145:classification / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D018533:Biomass; D050260:Carbohydrate Metabolism; D002480:Cellulase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D002850:Chromatography, Gel; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000737:chemistry / Q000235:genetics; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008969:Molecular Sequence Data; D008970:Molecular Weight; D032107:Populus / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013696:Temperature
chemical_list: D004271:DNA, Fungal; D021903:DNA, Ribosomal Spacer; D002480:Cellulase