title: Purification and characterization of an organic-solvent-tolerant cellulase from a halotolerant isolate, Bacillus sp. L1.
abstract: A halotolerant isolate Bacillus sp. L1 producing extracellular cellulase was isolated from Yuncheng, China. Production of the enzyme started from mid-exponential phase of bacterial growth and reached a maximum level during the post-stationary phase. The cellulase was purified to homogeneity with molecular mass of 45 kDa. Substrate specificity test indicated that it was an endoglucanase for soluble cellulose. Optimal enzyme activity was found to be at 60 °C, pH 8.0, and 7.5 % NaCl. Furthermore, it was highly active and stable over broad ranges of temperature (30-80 °C), pH (7.0-9.0), and NaCl concentration (2.5-15 %), thus showing its excellent thermostable, alkali-stable, and halotolerant nature. The cellulase activity was greatly inhibited by ethylenediaminetetraacetic acid, indicating that it was a metalloenzyme. Significant inhibition by phenylmethylsulfonyl fluoride and phenylarsine oxide revealed that serine and cysteine residues were essential for the enzyme catalysis. Moreover, the cellulase was highly active in the presence of surfactants, and it showed high stability in the presence of water-insoluble organic solvents with log P (ow)at least 0.88. Results from this study indicate that the purified cellulase from isolate L1 may have considerable potential for industrial application owing to its useful properties.
authors.affiliation: Life Science College, Yuncheng University, Yuncheng, 044000, China. lixin-eva@163.com
authors.forename: Xin
authors.initials: X
authors.lastname: Li
authors.forename: Hui-Ying
authors.initials: HY
authors.lastname: Yu
mesh_terms: D001407:Bacillus / Q000145:classification / Q000201:enzymology* / Q000254:growth & development / Q000302:isolation & purification; D055162:Biocatalysis / Q000187:drug effects; D002480:Cellulase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D002482:Cellulose / Q000378:metabolism; D002681:China; D004492:Edetic Acid / Q000494:pharmacology; D004795:Enzyme Stability / Q000187:drug effects; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D012965:Sodium Chloride / Q000494:pharmacology; D012997:Solvents / Q000737:chemistry / Q000494:pharmacology; D013379:Substrate Specificity; D013501:Surface-Active Agents / Q000494:pharmacology; D013696:Temperature
chemical_list: D012997:Solvents; D013501:Surface-Active Agents; D012965:Sodium Chloride; D002482:Cellulose; D004492:Edetic Acid; D002480:Cellulase