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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 |
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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.
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Received: 20 December 2013
Published: 25 March 2014
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