scholarly journals Identification of Phenol-Degrading Nocardia Sp. Strain C-14-1 and Characterization of Its Ring-Cleavage 2,3-Dioxygenase

2010 ◽  
Vol 2 (1) ◽  
Author(s):  
Haijuan Ma ◽  
Guangming Li ◽  
Ping Fang ◽  
Yalei Zhang ◽  
Deqiang Xu
Keyword(s):  
2005 ◽  
Vol 187 (15) ◽  
pp. 5067-5074 ◽  
Author(s):  
Daisuke Kasai ◽  
Eiji Masai ◽  
Keisuke Miyauchi ◽  
Yoshihiro Katayama ◽  
Masao Fukuda

ABSTRACT Sphingomonas paucimobilis SYK-6 converts vanillate and syringate to protocatechuate (PCA) and 3-O-methylgallate (3MGA) in reactions with the tetrahydrofolate-dependent O-demethylases LigM and DesA, respectively. PCA is further degraded via the PCA 4,5-cleavage pathway, whereas 3MGA is metabolized via three distinct pathways in which PCA 4,5-dioxygenase (LigAB), 3MGA 3,4-dioxygenase (DesZ), and 3MGA O-demethylase (LigM) are involved. In the 3MGA O-demethylation pathway, LigM converts 3MGA to gallate, and the resulting gallate appears to be degraded by a dioxygenase other than LigAB or DesZ. Here, we isolated the gallate dioxygenase gene, desB, which encodes a 418-amino-acid protein with a molecular mass of 46,843 Da. The amino acid sequences of the N-terminal region (residues 1 to 285) and the C-terminal region (residues 286 to 418) of DesB exhibited ca. 40% and 27% identity with the sequences of the PCA 4,5-dioxygenase β and α subunits, respectively. DesB produced in Escherichia coli was purified and was estimated to be a homodimer (86 kDa). DesB specifically attacked gallate to generate 4-oxalomesaconate as the reaction product. The Km for gallate and the V max were determined to be 66.9 ± 9.3 μM and 42.7 ± 2.4 U/mg, respectively. On the basis of the analysis of various SYK-6 mutants lacking the genes involved in syringate degradation, we concluded that (i) all of the three-ring cleavage dioxygenases are involved in syringate catabolism, (ii) the pathway involving LigM and DesB plays an especially important role in the growth of SYK-6 on syringate, and (iii) DesB and LigAB are involved in gallate degradation.


2012 ◽  
Vol 161 (2) ◽  
pp. 86-91 ◽  
Author(s):  
Yunior Morán ◽  
Osmani Chacón ◽  
Maria del Carmen Córdoba-Sellés ◽  
Rosario Domínguez-Larrinaga ◽  
Lidcay Herrera ◽  
...  

2000 ◽  
Vol 6 (2) ◽  
pp. 193-203 ◽  
Author(s):  
Masayuki Kubota ◽  
Keiichi Yoshida ◽  
Akira Tawada ◽  
Mamoru Ohashi

Positive- and negative-ion fast-atom bombardment tandem mass spectrometry with collision-induced dissociation (FAB-CID-MS/MS) has been used in the characterization of di-and tetra-saccharides of the lactosamine series from keratan sulfates. FAB-CID-MS/MS of Galβ1-4GlcNAc (L1) exhibited strong fragment ions originating from ring cleavage at the reducing-terminal sugar moiety together with glycosidic bond-cleavage ions, whereas GlcNAcβ1-3Gal (K1) showed strong glycosidic bond-cleavage ions but no ring-cleavage ions. A series of ring-cleavage fragment ions was observed with members of the L-series which have free hydroxyl groups at the C1 and C3 positions. CID-MS/MS spectra of the [M + Na – SO3]+ ion ( m/z 406) from L2 and the [M + Na − 2SO3]+ ion ( m/z 406) from L4 were almost identical with the CID-MS/MS spectrum of the [M + Na]+ ion ( m/z 406) from L1, which indicated that the sugar skeletons of L2 and L4 are the same as that of L1. On the other hand, the CID-MS/MS spectrum of the [M + Na – SO3]+ ion ( m/z 508) from L4 did not resemble that of the [M + Na]+ ion ( m/z 508) from L2. The former showed peaks that were additional to the peaks in the latter. Since these extra peaks were accounted for on the basis of the structure of L3 [Galβ1(6S)-4GlcNAc, S = sulfate], the in-source loss of sulfate groups by ester exchange upon FAB ionization takes place in a dual manner; one reaction at the non-reducing terminal sugar to give L2 and the other at the reducing-terminal sugar to give L3. The CID-MS/MS spectra were characteristic for the tetrasaccharides L1-L1, L2-L2 and L4-L4 while in-source fragmentation confirms the component disaccharides of each tetrasaccharide. The structure of a tetrasaccharide trisulfate was confirmed as L2–L4 and not L4–L2 by CID-MS/MS. Negative-ion FAB-CID-MS/MS spectra of the sulfated di-and tetra-saccharides showed a pattern similar to that of the positive-ion spectra. Subtraction of the CID-MS/MS spectrum of the [M – H]− ion of L2 [Galβ1-4GlcNAc(6S)] from that of the [M – H – SO3]− ion of L4 [Gal(6S)β1-4GlcNAc(6S)] gave several specific ions whose origins were nicely explained on the basis of the structure of L3. The structure of a pentasaccharide consisting of N-acetylneuraminic acid and a tetrasaccharide trisulfate was confirmed, on the basis of FAB-CID-MS/MS, as NeuNAcα2-6L2-L4.


2002 ◽  
Vol 68 (10) ◽  
pp. 4764-4771 ◽  
Author(s):  
Helena M. B. Seth-Smith ◽  
Susan J. Rosser ◽  
Amrik Basran ◽  
Emma R. Travis ◽  
Eric R. Dabbs ◽  
...  

ABSTRACT Hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) is a high explosive which presents an environmental hazard as a major land and groundwater contaminant. Rhodococcus rhodochrous strain 11Y was isolated from explosive contaminated land and is capable of degrading RDX when provided as the sole source of nitrogen for growth. Products of RDX degradation in resting-cell incubations were analyzed and found to include nitrite, formaldehyde, and formate. No ammonium was excreted into the medium, and no dead-end metabolites were observed. The gene responsible for the degradation of RDX in strain 11Y is a constitutively expressed cytochrome P450-like gene, xplA, which is found in a gene cluster with an adrenodoxin reductase homologue, xplB. The cytochrome P450 also has a flavodoxin domain at the N terminus. This study is the first to present a gene which has been identified as being responsible for RDX biodegradation. The mechanism of action of XplA on RDX is thought to involve initial denitration followed by spontaneous ring cleavage and mineralization.


2013 ◽  
Vol 97 (11) ◽  
pp. 5043-5053 ◽  
Author(s):  
Rabea Schlüter ◽  
Ramona Lippmann ◽  
Elke Hammer ◽  
Manuela Gesell Salazar ◽  
Frieder Schauer

2000 ◽  
Vol 31 (3) ◽  
pp. 249 ◽  
Author(s):  
Soon Han Kim ◽  
Ee Jong Lim ◽  
Sang Ok Lee ◽  
Jae Dong Lee ◽  
Tae Ho Lee

Author(s):  
Michael Haas ◽  
Roland Fischer ◽  
Ana Torvisco ◽  
Harald Stueger

Based on earlier work by Ottosson [1] and Ohshita [2] the previously unknown cyclic silenolates 1a-c and 2a-b have been synthesized by the reaction of acylcyclohexasilanes with one or two equivalents of KOtBu. The nature of the anions (silenide or silenolate) and consequently the outcome of subsequent reaction steps largely depends on the substituent R at the carbonyl function. Detailed NMR-analysis of 1a-c and 2a-b corroborate this deduction. Furthermore, for 1a-c and 2a-b single crystals which were suitable for X-ray structure analysis could be grown. Very interesting is the reactivity of 1b, which at 50°C undergoes a hitherto unknown 1,4-trimethylsilyl-migration followed by a ring cleavage of the cyclohexasilane cycle to form the anion 3a. The high selectivity of these reactions is demonstrated by the formation of an acyl-functionalized bicyclo-2,2,2-octasilane in >90% yield from 2a and \(ClSiMe_{2}SiMe_{2}Cl\).


1998 ◽  
Vol 180 (19) ◽  
pp. 5159-5164 ◽  
Author(s):  
G. Vijay Bhasker Reddy ◽  
Maarten D. Sollewijn Gelpke ◽  
Michael H. Gold

ABSTRACT Under secondary metabolic conditions, the lignin-degrading basidiomycete Phanerochaete chrysosporium mineralizes 2,4,6-trichlorophenol. The pathway for the degradation of 2,4,6-trichlorophenol has been elucidated by the characterization of fungal metabolites and oxidation products generated by purified lignin peroxidase (LiP) and manganese peroxidase (MnP). The multistep pathway is initiated by a LiP- or MnP-catalyzed oxidative dechlorination reaction to produce 2,6-dichloro-1,4-benzoquinone. The quinone is reduced to 2,6-dichloro-1,4-dihydroxybenzene, which is reductively dechlorinated to yield 2-chloro-1,4-dihydroxybenzene. The latter is degraded further by one of two parallel pathways: it either undergoes further reductive dechlorination to yield 1,4-hydroquinone, which isortho-hydroxylated to produce 1,2,4-trihydroxybenzene, or is hydroxylated to yield 5-chloro-1,2,4-trihydroxybenzene, which is reductively dechlorinated to produce the common key metabolite 1,2,4-trihydroxybenzene. Presumably, the latter is ring cleaved with subsequent degradation to CO2. In this pathway, the chlorine at C-4 is oxidatively dechlorinated, whereas the other chlorines are removed by a reductive process in which chlorine is replaced by hydrogen. Apparently, all three chlorine atoms are removed prior to ring cleavage. To our knowledge, this is the first reported example of aromatic reductive dechlorination by a eukaryote.


Sign in / Sign up

Export Citation Format

Share Document