title: Isolation and characterization of two novel halotolerant Catechol 2, 3-dioxygenases from a halophilic bacterial consortium.
abstract: Study of enzymes in halophiles will help to understand the mechanism of aromatic hydrocarbons degradation in saline environment. In this study, two novel catechol 2,3-dioxygenases (C23O1 and C23O2) were cloned and overexpressed from a halophilic bacterial consortium enriched from an oil-contaminated saline soil. Phylogenetic analysis indicated that the novel C23Os and their relatives formed a new branch in subfamily I.2.A of extradiol dioxygenases and the sequence differences were further analyzed by amino acid sequence alignment. Two enzymes with the halotolerant feature were active over a range of 0-30% salinity and they performed more stable at high salinity than in the absence of salt. Surface electrostatic potential and amino acids composition calculation suggested high acidic residues content, accounting for their tolerance to high salinity. Moreover, two enzymes were further characterized. The enzymes activity both increased in the presence of Fe(3+), Fe(2+), Cu(2+) and Al(3+) and showed no significant inhibition by other tested metal ions. The optimal temperatures for the C23Os were 40 °C and 60 °C and their best substrates were catechol and 4-methylcatechol respectively. As the firstly isolated and characterized catechol dioxygenases from halophiles, the two halotolerant C23Os presented novel characteristics suggesting their potential application in aromatic hydrocarbons biodegradation.
authors.affiliation: State Key Joint Laboratory of Environment Simulation and Pollution Control, School of Environment, Tsinghua University, Beijing, 100084, China.
authors.forename: Guang
authors.initials: G
authors.lastname: Guo
authors.affiliation: State Key Joint Laboratory of Environment Simulation and Pollution Control, School of Environment, Tsinghua University, Beijing, 100084, China.
authors.forename: Tingting
authors.initials: T
authors.lastname: Fang
authors.affiliation: State Key Joint Laboratory of Environment Simulation and Pollution Control, School of Environment, Tsinghua University, Beijing, 100084, China.
authors.forename: Chongyang
authors.initials: C
authors.lastname: Wang
authors.affiliation: State Key Joint Laboratory of Environment Simulation and Pollution Control, School of Environment, Tsinghua University, Beijing, 100084, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Huang
authors.affiliation: State Key Joint Laboratory of Environment Simulation and Pollution Control, School of Environment, Tsinghua University, Beijing, 100084, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Tian
authors.affiliation: State Key Joint Laboratory of Environment Simulation and Pollution Control, School of Environment, Tsinghua University, Beijing, 100084, China.
authors.forename: Qijia
authors.initials: Q
authors.lastname: Cui
authors.affiliation: State Key Joint Laboratory of Environment Simulation and Pollution Control, School of Environment, Tsinghua University, Beijing, 100084, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Wang
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D050557:Catechol 2,3-Dioxygenase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D004795:Enzyme Stability / Q000502:physiology; D019605:Euryarchaeota / Q000201:enzymology / Q000235:genetics; D006358:Hot Temperature; D059013:Microbial Consortia; D013379:Substrate Specificity / Q000502:physiology
chemical_list: D001426:Bacterial Proteins; D050557:Catechol 2,3-Dioxygenase