title: Properties of an ionic liquid-tolerant Bacillus amyloliquefaciens CMW1 and its extracellular protease.
abstract: An ionic liquid-tolerant bacterium, Bacillus amyloliquefaciens CMW1, was isolated from a Japanese fermented soybean paste. Strain CMW1 grew in the presence of 10 % (v/v) 1-butyl-3-methylimidazolium chloride ([BMIM]Cl), a commonly used ionic liquid. Additionally, strain CMW1 grew adequately in the presence of the hydrophilic ionic liquids 10 % (v/v) 1-ethyl-3-methylimidazolium trifluoromethanesulfonate ([EMIM]CF3SO3) or 2.5 % (v/v) 1-butyl-3-methylimidazolium trifluoromethanesulfonate ([BMIM]CF3SO3). Strain CMW1 produced an extracellular protease (BapIL) in the culture medium. BapIL was stable in the presence of 80 % (v/v) ionic liquids, [EMIM]CF3SO3, [BMIM]Cl, [BMIM]CF3SO3, 1-butyl-3-methylimidazolium tetrafluoroborate, 1-butyl-3-methylimidazolium hexafluorophosphate, and 1-butyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide, and functioned in 10 % (v/v) these ionic liquids. BapIL was stable at pH 4.0-12.6 or in 4004 mM NaCl solution, and exhibited activity in the presence of 50 % (v/v) hydrophilic or hydrophobic organic solvents. BapIL was completely inhibited by 1 mM PMSF and partially by 5 mM EDTA. BapIL belongs to the true subtilisins according to analysis of the deduced amino acid sequence. We showed that BapIL from the ionic liquid-tolerant B. amyloliquefaciens CMW1 exhibited tolerance to ionic liquid and halo, alkaline, and organic solvents.
authors.affiliation: Department of Applied Biological Chemistry, Faculty of Agriculture, Kinki University, 3327-204 Nakamachi, Nara, 631-8505, Japan. kurata090401@nara.kindai.ac.jp.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Kurata
authors.affiliation: Department of Applied Biological Chemistry, Faculty of Agriculture, Kinki University, 3327-204 Nakamachi, Nara, 631-8505, Japan.
authors.forename: Humiya
authors.initials: H
authors.lastname: Senoo
authors.affiliation: Department of Applied Biological Chemistry, Faculty of Agriculture, Kinki University, 3327-204 Nakamachi, Nara, 631-8505, Japan.
authors.forename: Yasuyuki
authors.initials: Y
authors.lastname: Ikeda
authors.affiliation: Department of Applied Biological Chemistry, Faculty of Agriculture, Kinki University, 3327-204 Nakamachi, Nara, 631-8505, Japan.
authors.forename: Hideaki
authors.initials: H
authors.lastname: Kaida
authors.affiliation: Department of Applied Biological Chemistry, Faculty of Agriculture, Kinki University, 3327-204 Nakamachi, Nara, 631-8505, Japan.
authors.forename: Chiaki
authors.initials: C
authors.lastname: Matsuhara
authors.affiliation: Department of Applied Biological Chemistry, Faculty of Agriculture, Kinki University, 3327-204 Nakamachi, Nara, 631-8505, Japan.
authors.forename: Noriaki
authors.initials: N
authors.lastname: Kishimoto
mesh_terms: D000069977:Bacillus amyloliquefaciens / Q000201:enzymology* / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D004795:Enzyme Stability / Q000187:drug effects; D052578:Ionic Liquids / Q000494:pharmacology*; D010447:Peptide Hydrolases / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D052578:Ionic Liquids; D010447:Peptide Hydrolases
keywords: B. amyloliquefaciens CMW1; Ionic Liquid; Ionic liquid-tolerant protease