Cloning and expression of a novel thermostable cellulase from newly isolated Bacillus subtilis strain I15
Tóm tắt
A novel bacterial strain with high cellulase activity (2.82 U/ml) was isolated, and then identified by its morphological character and 16S rRNA sequence, and named Bacillus subtilis strain I15. The extracellular thermostable cellulase exhibited the maximum activity at 60°C and pH 6.0. It was very stable since more than 90% of original CMCase activity was maintained at 65°C after incubation for 2 h. The cellulase gene, celI15, was cloned and extracellularly expressed by Escherichia coli BL21 (DE3), which encoded the extracellular protein of about 52 kDa. The extracellular activity of CelI15 from E. coli BL21 was up to about 6.78 U/ml, and all the other properties were almost the same as that from the wild-type strain.
Tài liệu tham khảo
Beguin P, Aubert JP (1994) The biological degradation of cellulose. FEMS Microbiol Rev 13:25–58. doi:10.1111/j.1574-6976.1994.tb00033.x
Ito S (1997) Alkaline cellulases from alkaliphilic Bacillus: enzymatic properties, genetics, and application to detergents. Extremophiles 1(2):61–66. doi:10.1007/s007920050015
Oksanen T, Poabilanina J (1998) Alkaline detergent enzymes from alkaliphile enzymtic properties, genetics and structures. Extremophiles 2(3):185–190. doi:10.1007/s007920050059
Blanco A, Diaz P, Martinez J, Vidal T, Torres AL, Pastor F (1998) Cloning of a new endoglucanase gene from Bacillus sp. BP-23 characterization of enzyme, performance in paper manufacture from cereal straw. Appl Microbiol Biotechnol 50:48–54. doi:10.1007/s002530051255
Doyle J, Pavel R, Barness G, Steinberger Y (2006) Cellulase dynamics in a desert soil. Soil Biol Biochem 38:371–376
Li YH, Ding M, Wang J, Xu GJ, Zhao F (2006) A novel thermoacidophilic endoglucanase, Ba-EGA, from a new cellulose-degrading bacterium, Bacillus sp. AC-1. Appl Microbiol Biotechnol 70:430–436. doi:10.1007/s00253-005-0075-x
Paula SP, Alexandra M, Jose CD, Maria R, Aires B, Maria CF (2002) Rapid production of thermostable cellulase free xylanase by a strain of Bacillus subtilis and its properties. Enzyme Microb Technol 30:924–933. doi:10.1016/S0141-0229(02)00034-0
Li X, Gao P (1982) Isolation and partial properties of cellulose-decomposing strain of Cytophaga LX-7 from soil. Appl Environ Microbiol 38:148–158
Nazneen B, Javed M (2003) Characterization of a new Pseudomonas aeruginosa strain NJ-15 as a potential biocontrol agent. Curr Microbiol 46:324–328. doi:10.1007/s00284-002-3857-8
Miller GL (1959) Use of dinitrosalicylic acid reagent for the determination of reducing sugar. Anal Chem 31:426–428. doi:10.1021/ac60147a030
Kim CH, Kim DS (1992) Production and characterization of crystalline cellulose-degrading cellulase components from a thermophilic and moderately alkalophilic bacterium. J Microbiol Biotechnol 2:7–13
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold SpringHarbor Laboratory Press, New York
Yoshihiro H, Kenzo K, Tadashi Y, Hajimi M, Tohru K, Susumu I (1997) Thermostable alkaline cellulase from an alkaliphilic isolate, Bacillus sp. KSM-S237. Extremophiles 1:151–156. doi:10.1007/s007920050028
Robertson LD, Koehn RD (1978) Characteristics of the cellulase produced by the ascomycete Poronia punctata. Mycologia 70:1113–1121. doi:10.2307/3759142
Li W, Zhang WW, Yang MM, Chen YL (2008) Cloning of the thermostable cellulase gene from newly isolated Bacillus subtilis and its expression in Escherichia coli. Mol Biotechnol 40(2):195–201. doi:10.1007/s12033-008-9079-y
Thongekkaew J, Ikeda H, Masaki K, Iefuji H (2008) An acidic and thermostable carboxymethyl cellulase from the yeast Cryptococcus sp. S-2: purification, characterization and improvement of its recombinant enzyme production by high cell-density fermentation of Pichia pastoris. Protein Expr Purif 60(2):140–146. doi:10.1016/j.pep.2008.03.021
Aphichart K, Amorn P, Polkit S (2008) A novel thermostable endoglucanase from the wood-decaying fungus Daldinia eschscholzii (Ehrenb.:Fr.) Rehm. Enzym Microb Technol 42:404–413. doi:10.1016/j.enzmictec.2007.11.009
Benjamaporn W, Kusol P, Katewadee B (2008) Cloning, expression and characterization of a thermotolerant endoglucanase from Syncephalastrum racemosum (BCC18080) in Pichia pastoris. Protein Expr Purif 58:78–86. doi:10.1016/j.pep.2007.10.022
Aa K, Flengsrud R, Lindahl V, Tronsmo A (1994) Characterization of production and enzyme properties of an endo-beta-1,4-glucanase from Bacillus subtilis CK-2 isolated from compost soil. Antonie Van Leeuwenhoek 66(4):319–326. doi:10.1007/BF00882767
Arnold K, Bordoli L, Kopp J, Schwede T (2006) The SWISS-MODEL workspace: a web-based environment for protein structure homology modeling. Bioinformatics 22:195–201. doi:10.1093/bioinformatics/bti770
Schwede T, Kopp J, Guex N, Peitsch MC (2003) SWISS-MODEL: an automated protein homology-modeling server. Nucleic Acids Res 31:3381–3385. doi:10.1093/nar/gkg520
Guex N, Peitsch MC (1997) SWISS-MODEL and the swiss-Pdb viewer: an environment for comparative protein modelling. Electrophoresis 18:2714–2723. doi:10.1002/elps.1150181505
Sonan GK, Receveur-Brechot V, Duez C, Aghajari N, Czjzek M, Haser R, Gerday C (2007) The linker region plays a key role in the adaptation to cold of the cellulase from an Antarctic bacterium. Biochem J 407(2):293–302. doi:10.1042/BJ20070640
