title: A novel efficient β-glucanase from a paddy soil microbial metagenome with versatile activities.
abstract: BACKGROUND
  Cellulose, an abundant and renewable polysaccharides, constitutes the largest resource for bioconversion of biofuels. Plant polysaccharides hydrolysis is catalyzed by cellulases, which include endoglucanases, exoglucanases, and β-glucosidases. Converting cellulose and hemicellulose to short chains of oligosaccharides by endo-/exoglucanases is the key step for biofuel transformation. Intriguingly, β-glucanases with transglycosylation activity not only can relieve product inhibition of glucan hydrolysis but also has potential application as biocatalysts for functional materials.
  
  
  RESULTS
  Here, a metagenomic fosmid library was constructed from a paddy soil for cellulase screening. One purified clone showing carboxymethylcellulase activity was isolated, and the complete β-glucanase gene (umcel9y-1) was cloned and overexpressed in Escherichia coli. Phylogenetic analysis indicated that β-glucanase Umcel9y-1 belonged to the theme C of glycoside hydrolase family 9. Amino acids sequence showed 58.4 % similarity between Umcel9y-1 and its closest characterized reference, cellulase Cel01. Biological characterization showed that Umcel9y-1 was an efficient endoglucanase and also exhibited high activities of exoglucanase and transglycosylation. The transglycosylation products of Umcel9y-1 including sophorose, laminaribiose, and gentiobiose, and transglycosylation was detected under all activated conditions. The order of catalytic efficiency for polysaccharides, cellooligosaccharides, and aryl-β-glycosides was p-nitrophenol-D-cellobioside, barley glucan, cellopentaose, cellotetraose, cellotriose, hydroxyethylcellulose, cellohexose, laminarin, and carboxymethylcellulose, respectively. The barley glucan was the optimal polysaccharides for Umcel9y-1 with K m and K cat/K m values of 13.700 mM and 239.152 s(-1) mM(-1), respectively.
  
  
  CONCLUSIONS
  Biological characterizations of recombinant Umcel9y-1 showed that the versatile β-glucanase had efficient endoglucanase activity to barley glucan and also exhibited high activities of exoglucanase and transglycosylation. The optimum conditions of recombinant Umcel9y-1 was pH 6.5-7.0 at 37 °C with predominant halotolerance and high-thermal stability. These results indicate that the novel metagenomic-derived β-glucanase may be a potent candidate for industrial applications.
authors.affiliation: State Key Laboratory of Tea Biology and Utilization, School of Tea and Food Science Technology, Anhui Agricultural University, Hefei, 230036 China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: State Key Laboratory of Tea Biology and Utilization, School of Tea and Food Science Technology, Anhui Agricultural University, Hefei, 230036 China.
authors.forename: Xu
authors.initials: X
authors.lastname: Wang
authors.affiliation: Institute of Quality and Standard for Agro-products, Zhejiang Academy of Agricultural Sciences, Hangzhou, 310021 China.
authors.forename: Wei
authors.initials: W
authors.lastname: Wei
authors.affiliation: Institute of Quality and Standard for Agro-products, Zhejiang Academy of Agricultural Sciences, Hangzhou, 310021 China.
authors.forename: Jimin
authors.initials: J
authors.lastname: Xu
authors.affiliation: Institute of Quality and Standard for Agro-products, Zhejiang Academy of Agricultural Sciences, Hangzhou, 310021 China.
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Tea Biology and Utilization, School of Tea and Food Science Technology, Anhui Agricultural University, Hefei, 230036 China.
authors.forename: Zhongwen
authors.initials: Z
authors.lastname: Xie
authors.affiliation: State Key Laboratory of Tea Biology and Utilization, School of Tea and Food Science Technology, Anhui Agricultural University, Hefei, 230036 China.
authors.forename: Zhengzhu
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Biogeology and Environmental Geology, China University of Geosciences, Wuhan, 430074 China.
authors.forename: Hongchen
authors.initials: H
authors.lastname: Jiang
authors.affiliation: Novus International (Shanghai) Inc, Shanghai, 200080 China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Tea Biology and Utilization, School of Tea and Food Science Technology, Anhui Agricultural University, Hefei, 230036 China.
authors.forename: Chaoling
authors.initials: C
authors.lastname: Wei
keywords: Endoglucanase; Exoglucanases; Glycoside hydrolase family 9; Metagenomic library; Transglycosylation; Versatile β-glucanase