TAILIEUCHUNG - Kolvenbach et al. AMB Express 2011, 1:8 http://www.amb-express.com/content/1/1/8 ORIGINAL Open

Kolvenbach et al. AMB Express 2011, 1:8 ORIGINAL Open Access Purification and characterization of hydroquinone dioxygenase from Sphingomonas sp. strain TTNP3 Boris A Kolvenbach1,2*, Markus Lenz1, Dirk Benndorf3, Erdmann Rapp4, Jan Fousek5,6, Cestmir Vlcek5,6, Andreas Schäffer2, Frédéric LP Gabriel7, Hans-Peter E Kohler8 and Philippe FX Corvini1,9 Abstract Hydroquinone-1,2-dioxygenase, an enzyme involved in the degradation of alkylphenols in Sphingomonas sp. strain TTNP3 was purified to apparent homogeneity. The extradiol dioxygenase catalyzed the ring fission of hydroquinone to 4-hydroxymuconic semialdehyde and the degradation of chlorinated and several alkylated hydroquinones. The activity of 1 mg of the purified enzyme with unsubstituted hydroquinone was μmol per minute,. | Kolvenbach et al. AMB Express 2011 1 8 http content 1 1 8 o AMB Express a SpringerOpen Journal ORIGINAL Open Access Purification and characterization of hydroquinone dioxygenase from Sphingomonas sp. strain TTNP3 Dwrir A If IV ỉ K s r h 1 2K ỉ V 1 1 I r I 71 Ph I D Z XV I Kp3 p rh D s k 4 I p I I r I 5 6 r f- I V I r I 5 6 oris A Koivenoach MarKus Lenz DirK Bennoorr Erdmann Rapp Jan FouseK Cestmir vlceK AnHrosỉc BAh affor2 BroHorA I D fBAKriol7 B kAhlor8 f lh Bhilir r ci BV r trx ini1 9 Andreas jChdiie Frederic LP Gabriel nans Peter t Kohler ano Philippe FA Corvini Abstract nydroquinone-1 2-dioxygenase an enzyme involved in the degradation of alKylphenols in Sphingomonas sp. strain TTNP3 was purified to apparent homogeneity. The extradiol dioxygenase catalyzed the ring fission of hydroquinone to 4-hydroxymuconic semialdehyde and the degradation of chlorinated and several alKylated hydroquinones. The activity of 1 mg of the purified enzyme with unsubstituted hydroquinone was gmol per minute the apparent Km gM. ICP-MS analysis revealed an iron content of moles per mole enzyme. The enzyme lost activity upon exposure to oxygen but could be reactivated by Fe II in presence of ascorbate. SDS-PAGt analysis of the purified enzyme yielded two bands of an apparent size of 38 KDa and 19 KDa respectively. Data from MALDI-TOF analyses of peptides of the respective bands matched with the deduced amino acid sequences of two neighboring open reading frames found in genomic DNA of Sphingomonas sp strain TTNP3. The deduced amino acid sequences showed 62 and 47 identity to the large and small subunit of hydroquinone dioxygenase from Pseudomonas fluorescens strain ACB respectively. This heterotetrameric enzyme is the first of its Kind found in a strain of the genus Sphingomonas sensu latu. Keywords hydroquinone dioxygenase Sphingomonas nonylphenol bisphenol A Introduction Both Sphingomonas sp. strain TTNP3 and Sphingobium xenophagum Bayram are able to .

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