title: Homologous cloning, purification and characterization of highly active cellobiohydrolase I (Cel7A) from Penicillium canescens.
abstract: Penicillium canescens is a filamentous fungus that normally does not secrete notable levels of cellulase activity. Cellobiohydrolase I of P. canescens (PcCel7A) was homologously cloned into a host strain RN3-11-7 (niaD-) and then expressed under the control of a strong xylA promoter. Using three steps of chromatography, PcCel7A was purified. The enzyme displayed maximum activity at pH 4.0-4.5. PcCel7A was stable at 50°C and pH 4.5 at least for 3h, while at 60°C it lost 45% of activity after 30min of incubation. When equalized by protein concentration, PcCel7A demonstrated a higher performance in prolonged hydrolysis of Avicel and milled aspen wood than CBH I (Cel7A) from Trichoderma reesei, the most industrially utilized cellulase at this moment. The high catalytic efficiency of the PcCel7A makes it a potential candidate for industrial applications.
authors.affiliation: A. N. Bach Institute of Biochemistry, Russian Academy of Sciences, Leninsky Pr. 33, Moscow 119071, Russia.
authors.forename: Pavel V
authors.initials: PV
authors.lastname: Volkov
authors.affiliation: A. N. Bach Institute of Biochemistry, Russian Academy of Sciences, Leninsky Pr. 33, Moscow 119071, Russia.
authors.forename: Alexandra M
authors.initials: AM
authors.lastname: Rozhkova
authors.affiliation: A. N. Bach Institute of Biochemistry, Russian Academy of Sciences, Leninsky Pr. 33, Moscow 119071, Russia; Department of Chemistry, M. V. Lomonosov Moscow State University, Vorobyovy Gory 1/11, Moscow 119991, Russia. Electronic address: avgusakov@enzyme.chem.msu.ru.
authors.forename: Alexander V
authors.initials: AV
authors.lastname: Gusakov
authors.affiliation: A. N. Bach Institute of Biochemistry, Russian Academy of Sciences, Leninsky Pr. 33, Moscow 119071, Russia; Department of Chemistry, M. V. Lomonosov Moscow State University, Vorobyovy Gory 1/11, Moscow 119991, Russia.
authors.forename: Arkady P
authors.initials: AP
authors.lastname: Sinitsyn
mesh_terms: D002482:Cellulose / Q000737:chemistry; D043366:Cellulose 1,4-beta-Cellobiosidase / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*; D003001:Cloning, Molecular; D006868:Hydrolysis; D010407:Penicillium / Q000201:enzymology*; D014242:Trichoderma / Q000201:enzymology; D014934:Wood / Q000737:chemistry
chemical_list: D002482:Cellulose; D043366:Cellulose 1,4-beta-Cellobiosidase
keywords: Cellobiohydrolase; Cellulase; Cloning; Penicillium canescens; Purification