Stereoselectivity and Conformational Stability of Haloalkane Dehalogenase DbjA from Bradyrhizobium japonicum USDA110: The Effect of pH and Temperature.

CHALOUPKOVÁ, Radka, Zbyněk PROKOP, Yukari SATO, Yuji NAGATA and Jiří DAMBORSKÝ. Stereoselectivity and Conformational Stability of Haloalkane Dehalogenase DbjA from Bradyrhizobium japonicum USDA110: The Effect of pH and Temperature. FEBS Journal. 2011, vol. 278, No 15, p. 2728-2738. ISSN 1742-464X. Available from: https://dx.doi.org/10.1111/j.1742-4658.2011.08203.x.

Basic information

• Original name: Stereoselectivity and Conformational Stability of Haloalkane Dehalogenase DbjA from Bradyrhizobium japonicum USDA110: The Effect of pH and Temperature.
• Authors: CHALOUPKOVÁ, Radka (203 Czech Republic, belonging to the institution), Zbyněk PROKOP (203 Czech Republic, belonging to the institution), Yukari SATO (392 Japan, belonging to the institution), Yuji NAGATA (392 Japan, belonging to the institution) and Jiří DAMBORSKÝ (203 Czech Republic, guarantor, belonging to the institution).
• Edition: FEBS Journal, 2011, 1742-464X.

Other information

• Original language: English
• Type of outcome: Article in a journal
• Field of Study: 10600 1.6 Biological sciences
• Country of publisher: United Kingdom of Great Britain and Northern Ireland
• Confidentiality degree: is not subject to a state or trade secret
• Impact factor: Impact factor: 3.790
• RIV identification code: RIV/00216224:14310/11:00049446
• Organization unit: Faculty of Science
• Doi: http://dx.doi.org/10.1111/j.1742-4658.2011.08203.x
• UT WoS: 000292933300011
• Keywords in English: DbJA; Enantioselectivity;Haloalkane Dehalogenase
• Tags: AKR, rivok

Abstract

The effect of pH and temperature on structure, stability, activity and enantioselectivity of haloalkane dehalogenase DbjA from Bradyrhizobium japonicum USDA110 was investigated in this study. Conformational changes have been assessed by circular dichroism spectroscopy, functional changes by kinetic analysis, while quaternary structure was studied by gel filtration chromatography. Our study shows that the DbjA enzyme is highly tolerant to pH changes. Its secondary and tertiary structure was not affected by pH in the range of 5.3-10.3 and 6.2-10.1, respectively. Oligomerization of DbjA was strongly pH-dependent: monomer, dimer, tetramer and a high molecular weight cluster of the enzyme was distinguished in solution at different pH conditions. Moreover, different oligomeric states of DbjA possessed different thermal stability.

Links

• GA203/08/0114, research and development project: Name: Specifické iontové efekty pro proteiny v roztocích a podobné biologicky relevantní systémy.

Investor: Czech Science Foundation, Specific ion effects for proteins in solutions and related biologically relevant systems

• IAA401630901, research and development project: Name: Evoluce substrátové specifity u enzymů aktivních s xenobiotickými látkami

Investor: Academy of Sciences of the Czech Republic, Evolution of substrate specificity in enzymes acting on xenobiotic compounds

• LC06010, research and development project: Name: Centrum biokatalýzy a biotransformací

Investor: Ministry of Education, Youth and Sports of the CR, Center of Biocatalysis and Biotransformation

• MSM0021622412, plan (intention): Name: Interakce mezi chemickými látkami, prostředím a biologickými systémy a jejich důsledky na globální, regionální a lokální úrovni (INCHEMBIOL) (Acronym: INCHEMBIOL)

Investor: Ministry of Education, Youth and Sports of the CR, Interactions among the chemicals, environment and biological systems and their consequences on the global, regional and local scales (INCHEMBIOL)