|
|
|
| Characterization of Cellulase Solution Produced by Leptosphaerulina chartarum SJTU59 and Analysis of Conserved Regions of Cellulase Genes |
| WU Qiong, WANG Meng, LI Ya-Qian, GAO Shi-Gang, YU Chuan-Jin, SUN Jia-Nan, ZHANG Tai-Long, CHEN Jie |
| Laboratory of Urban Agriculture(South)of Ministry of Agriculture, School of Agriculture and Biology, Shanghai Jiaotong University, Shanghai 200240, China |
|
|
|
Abstract Leptosphaerulina chartarum SJTU59 was firstly detected to produce cellulase solution in corn stalk powder via DNS (3, 5-Dinitrosalicylic acid) method. Under the optimal reaction temperature (50℃) and the optimal reaction pH (4.8), the enzyme activity of the cellulase solution was up to 6.383 ± 0.196 U/ml. The enzyme solution was relatively stable over a range of pH (pH 4.0 to pH 6.0) and temperature (20℃ to 60℃) while showing resistance to some metal ions tested. Two sets of degenerate primers were used for PCR amplification of the conserved regions of novel cellulase genes from L. chartarum SJTU59, named glu-l1 (1 512 bp) and glu-l2 (429 bp). Via the homology analysis of amino acid sequence and phylogenetic analysis, GLU-L1 which has 62% homology with β-1, 4-glucanases (ABX79553) from Thermoascus aurantiacus var. levisporus were inferred that a conserved region of cellulase of the glycoside hydrolase (GH) family 3, while GLU-L2 which has 49% homology with β-1, 3 (4)-glucanases (XP_755769) from Aspergillus fumigatus Af293 were inferred that a conserved region of cellulase of the glycoside hydrolase (GH) family 16.
|
|
Received: 20 December 2013
Published: 25 March 2014
|
|
|
|
|
[1] 孟雷, 陈冠军, 王怡, 等. 纤维素酶的多样性. 纤维素科学与技术, 2002, 10(2): 47-55. Meng L, Chen G J, Wang Y, et al. The polymorphism of cellulases. Journal of Cellulose Science and Technology, 2002, 10(2): 47-55.
[2] 郑亚平, 余晓斌. 碳源对绿色木霉ZC产中性纤维素酶的影响. 无锡轻工大学学报(食品与生物技术), 2003, 22(2): 30-33. Zheng Y P, Yu X B. The effect of carbon source for neutral cellulase produced by Trichoderma viride. Journal of Wuxi University of Light Industry, 2003, 22(2): 30-33.
[3] Tilbeurgh V H, Tomme P, Claeyssens M, et al. Limited proteolysis of the cellobiohydrolase I from Trichoderma reesei. FEBS Letters, 1986, 204(2): 223-227.
[4] 肖春玲, 徐常新. 微生物纤维素酶的应用研究. 微生物学杂志, 2002, 22(2): 33-35. Xiao C L, Xu C X. Application study on microbial cellulase. Journal of Microbiology, 2002, 22(2): 33-35.
[5] Wang S Y, Hu W, Lin X Y, et al. A novel cold-active xylanase from the cellulolytic myxobacterium Sorangium cellulosum So9733-1: gene cloning, expression, and enzymatic characterization. Applied Microbiology and Biotechnology, 2012, 93(4): 1503-1512.
[6] Yan Q J, Hua C W, Yang S Q, et al. High level expression of extracellular secretion of a β-glucosidase gene (PtBglu3) from Paecilomyces thermophila in Pichia pastoris. Protein Expression and Purification, 2012, 84(1): 64-72.
[7] Hua C W, Yan Q J, Jiang Z Q, et al. High-level expression of a specific β-1, 3-1, 4-glucanase from the thermophilic fungus Paecilomyces thermophila in Pichia pastoris. Applied Microbiology and Biotechnology, 2010, 88(2): 509-518.
[8] Tamura K, Dudley J, Nei M, et al. MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Molecular Biology and Evolution, 2007, 24(8): 1596-1599.
[9] 曾胤新, 俞勇, 陈波, 等. 低温纤维素酶产生菌的筛选、鉴定、生长特性及酶学性质. 高技术通讯, 2005, 15(4): 58-62. Zeng Y X, Yu Y, Chen B, et al. Screening, classification and growth of bacterium producingcold-active cellulase and its enzymic characterization. High Technology Letters, 2005, 15(4): 58-62.
[10] Hou Y H, Wang T H, Long H, et al. Novel cold-adaptive Penicillium strain FS010 secreting thermo-labile xylanase isolated from Yellow Sea. Acta Biochimica et Biophysica Sinica, 2006, 38(2): 142-149. |
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
