title: Characterization of endoglucanase from Paenibacillus sp. M33, a novel isolate from a freshwater swamp forest.
abstract: The newly isolated Paenibacillus sp. M33 from freshwater swamp forest soil in Thailand demonstrated its potential as a cellulose degrader. One of its endoglucanase genes from Paenibacillus sp., celP, was cloned to study the molecular characteristics of its gene product. The celP gene was recognized firstly by degenerate primer designed from Paenibacillus endoglucanase gene, and subsequently identified flanking region by inverse PCR technique. The celP gene consists of an open reading frame of 1707 bp encoding for 569 amino acids including 33-amino acids signal sequence. CelP is a member of glycoside hydrolase family 5 appended with a family 46 carbohydrate-binding module. CelP from recombinant Escherichia coli was purified by affinity chromatography. SDS-PAGE analysis of purified CelP showed a protein band at about 60 kDa. The purified enzyme gave a specific CMCase activity of 0.03 μmol min-1mg-1. It had higher activities on lichenan (0.19 μmol min-1mg-1) and barley β-glucan (0.14 μmol min-1mg-1). Maximum activity on lichenan was obtained at 50 °C, pH 5.0. CelP was stable over a pH range of 3.0-10.0 and retained 80% activity when incubated at 50 °C for 1 h. The properties of its CelP endoglucanase, especially substrate specificity, will make it useful in various biotechnological applications including biomass hydrolysis.
authors.affiliation: Faculty of Pharmacy, Department of Microbiology, Mahidol University, Bangkok, Thailand.
authors.forename: Pimpikar
authors.initials: P
authors.lastname: Kanchanadumkerng
authors.affiliation: Applied Microbiology Laboratory, Graduate School of Bioresources, Mie University, Mie, Japan.
authors.forename: Makiko
authors.initials: M
authors.lastname: Sakka
authors.affiliation: Applied Microbiology Laboratory, Graduate School of Bioresources, Mie University, Mie, Japan.
authors.forename: Kazuo
authors.initials: K
authors.lastname: Sakka
authors.affiliation: Faculty of Pharmacy, Department of Microbiology, Mahidol University, Bangkok, Thailand.
authors.forename: Chanpen
authors.initials: C
authors.lastname: Wiwat
mesh_terms: D000595:Amino Acid Sequence; D015373:Bacterial Typing Techniques; D001483:Base Sequence; D002480:Cellulase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D016000:Cluster Analysis; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004783:Environmental Microbiology; D004795:Enzyme Stability; D015870:Gene Expression; D005936:Glucans / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016366:Open Reading Frames; D056507:Paenibacillus / Q000145:classification / Q000201:enzymology* / Q000302:isolation & purification; D010802:Phylogeny; D021382:Protein Sorting Signals; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017422:Sequence Analysis, DNA; D013379:Substrate Specificity; D013696:Temperature; D013785:Thailand; D047071:beta-Glucans / Q000378:metabolism
chemical_list: D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D005936:Glucans; D021382:Protein Sorting Signals; D012336:RNA, Ribosomal, 16S; D011994:Recombinant Proteins; D047071:beta-Glucans; D002480:Cellulase; C015968:lichenin
keywords: Cellulase; Endoglucanase; Freshwater swamp forest; Glycoside hydrolase family 5; Paenibacillus sp.