HATTA, Takashi, Gouri MUKERJEE-DHAR, Jiri DAMBORSKY, Hohzoh KIYOHARA a Kazuhide KIMBARA. Cloning and characterization of a novel thermostable Mn(II)-dependent 2,3-dihydroxybiphenyl 1,2-dioxygenase from a PCB and naphthalene degrading Bacillus sp. JF8. Journal of Biological Chemistry, 2003, roč. 278, č. 24, s. 21483-21492. ISSN 0021-9258.
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Základní údaje
Originální název Cloning and characterization of a novel thermostable Mn(II)-dependent 2,3-dihydroxybiphenyl 1,2-dioxygenase from a PCB and naphthalene degrading Bacillus sp. JF8
Autoři HATTA, Takashi (392 Japonsko), Gouri MUKERJEE-DHAR (392 Japonsko), Jiri DAMBORSKY (203 Česká republika, garant), Hohzoh KIYOHARA (392 Japonsko) a Kazuhide KIMBARA (392 Japonsko).
Vydání Journal of Biological Chemistry, 2003, 0021-9258.
Další údaje
Originální jazyk angličtina
Typ výsledku Článek v odborném periodiku
Obor 1.6 Biological sciences
Stát vydavatele Spojené státy americké
Utajení není předmětem státního či obchodního tajemství
WWW URL
Impakt faktor Impact factor: 6.482
Kód RIV RIV/00216224:14310/03:00008775
Organizační jednotka Přírodovědecká fakulta
UT WoS 000183354200023
Klíčová slova anglicky ENZYME; THERMOSTABILITY; PCB; BIODEGRADATION; CLONING
Štítky biodegradation, CLONING, Enzyme, PCB, THERMOSTABILITY
Příznaky Recenzováno
Změnil Změnil: prof. Mgr. Jiří Damborský, Dr., učo 1441. Změněno: 19. 3. 2010 10:54.
Anotace
A novel thermostable Mn(II)-dependent 2,3-dihydroxybiphenyl 1,2-dioxygenase (BphC_JF8) catalyzing the meta-cleavage of the hydroxylated biphenyl ring was purified from the thermophilic biphenyl and naphthalene degrader, Bacillus sp. JF8 and the gene cloned. The native and recombinant BphC enzyme was purified to homogeneity. The enzyme has a molecular mass of 125+-10 kDa and was composed of 4 identical subunits (35 kDa). BphC_JF8 has a temperature optimum of 85 C and a pH optimum of 8.5. It exhibited a half life of 30 min at 80 C and 81 min at 75 C, making it the most thermostable extradiol dioxygenase studied. ICP-MS analysis confirmed the presence of 4.0 to 4.8 manganese atoms per enzyme molecule. The ESR spectrum of BphC_JF8 exhibited g = 2.02 and g = 4.06 signals having the 6-fold hyperfine splitting characteristic of Mn(II). The enzyme can oxidize a wide range of substrates and the substrate preference was in the order 2,3-dihydroxybiphenyl > 3-methylcatechol > catechol > 4-methylcatechol > 4-chlorocatechol. The enzyme is resistant to denaturation by various chelators and inhibitors (EDTA, 1,10-phenanthroline, H2O2, 3-chlorocatechol) and did not exhibit substrate inhibition even at 3 mM 2,3-dihydroxybiphenyl. A decrease in Km accompanied an increase in temperature and the Km value of 0.095 uM for 2,3-dihydroxybiphenyl (at 60 C) is among the lowest reported. The kinetic properties and thermal stability of the native and recombinant enzyme were identical. The primary structure of BphC_JF8 exhibits less than 25% sequence identity to other 2,3-dihydroxybiphenyl 1,2-dioxygenases. The metal ligands and active site residues of extradiol dioxygenases are conserved although several amino acid residues found exclusively in enzymes which preferentially cleave bicyclic substrates are missing in BphC_JF8. A 3-D homology model of BphC_JF8 provided a basis for understanding the substrate specificity, quaternary structure and stability of the enzyme.
Návaznosti
LN00A016, projekt VaVNázev: BIOMOLEKULÁRNÍ CENTRUM
Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Výzkumná centra
VytisknoutZobrazeno: 21. 5. 2019 03:29