scholarly journals 2,3-Dihydroxybenzoate pathway in Pseudomonas putida. 1H n.m.r. study on the ring-cleavage site

1981 ◽  
Vol 194 (2) ◽  
pp. 607-610 ◽  
Author(s):  
V Andreoni ◽  
L Canonica ◽  
E Galli ◽  
C Gennari ◽  
V Treccani
Keyword(s):  
Fission Product ◽  
Pseudomonas Putida ◽  
Cleavage Site ◽  
Dienoic Acid ◽  
Ring Cleavage ◽  
Ring Fission

1. Ring cleavage of 2,3-dihydroxybenzoate by cell-free extracts of Pseudomonas putida leads to 2-hydroxy-6-oxo-(2Z,4E)-hexa-2,4-dienoic acid and CO2. 2. The 1H n.m.r. spectrum of the ring-fission product obtained in a 2H2O solution suggests that the extra-diol cleavage occurs between C-3 and C-4.

scholarly journals Identification of a Serine Hydrolase Which Cleaves the Alicyclic Ring of Tetralin

2000 ◽  
Vol 182 (19) ◽  
pp. 5448-5453 ◽  
Author(s):  
M. J. Hernáez ◽  
E. Andújar ◽  
J. L. Ríos ◽  
S. R. Kaschabek ◽  
W. Reineke ◽  
...  
Keyword(s):  
Fission Product ◽  
Sequence Comparison ◽  
Dienoic Acid ◽  
Thermostable Enzyme ◽  
Insertion Mutant ◽  
Comparison Analysis ◽  
Serine Hydrolase ◽  
Carbon Carbon Bond ◽  
Ring Fission ◽  
Optimum Reaction

ABSTRACT A gene designated thnD, which is required for biodegradation of the organic solvent tetralin by Sphingomonas macrogoltabidus strain TFA, has been identified. Sequence comparison analysis indicated that thnD codes for a carbon-carbon bond serine hydrolase showing highest similarity to hydrolases involved in biodegradation of biphenyl. An insertion mutant defective in ThnD accumulates the ring fission product which results from the extradiol cleavage of the aromatic ring of dihydroxytetralin. The gene product has been purified and characterized. ThnD is an octameric thermostable enzyme with an optimum reaction temperature at 65°C. ThnD efficiently hydrolyzes the ring fission intermediate of the tetralin pathway and also 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid, the ring fission product of the biphenylmeta-cleavage pathway. However, it is not active towards the equivalent intermediates of meta-cleavage pathways of monoaromatic compounds which have small substituents in C-6. When ThnD hydrolyzes the intermediate in the tetralin pathway, it cleaves a C-C bond comprised within the alicyclic ring of tetralin instead of cleaving a linear C-C bond, as all other known hydrolases ofmeta-cleavage pathways do. The significance of this activity of ThnD for the requirement of other activities to mineralize tetralin is discussed.

scholarly journals The metabolism of thymol by a Pseudomonas

1968 ◽  
Vol 110 (4) ◽  
pp. 755-763 ◽  
Author(s):  
Enid M. Chamberlain ◽  
S. Dagley
Keyword(s):  
Fission Product ◽  
Pseudomonas Putida ◽  
Exponential Growth ◽  
Hydroxyl Group ◽  
Enzymic Hydrolysis ◽  
Ferrous Ions ◽  
Benzene Nucleus ◽  
Ring Fission ◽  
Hydrolysis Of

1. Pseudomonas putida when grown with thymol contained a meta-fission dioxygenase, which required ferrous ions and readily cleaved the benzene nucleus of catechols between adjacent carbon atoms bearing hydroxyl and isopropyl groups. 2. 3-Hydroxythymo-1,4-quinone was excreted towards the end of exponential growth and later was slowly metabolized. This compound was oxidized by partially purified extracts only when NADH was supplied; the substrate for the dioxygenase appeared to be 3-hydroxythymo-1,4-quinol, which was readily and non-enzymically oxidized to the quinone. 3. 2-Oxobutyrate (0·9 mole) was formed from 1 mole of 3-hydroxythymo-1,4-quinone with the consumption of 1 mole of oxygen; acetate, isobutyrate and 2-hydroxybutyrate (which arose from the enzymic reduction of 2-oxobutyrate) were also formed. 4. These products, which were produced only when the catechol substrate contained a third hydroxyl group, appeared to result from the enzymic hydrolysis of the ring-fission product.

scholarly journals Metabolism of biphenyl. Structure and physicochemical properties of 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid, the meta-cleavage product from 2,3-dihydroxybiphenyl by Pseudomonas putida

1974 ◽  
Vol 143 (2) ◽  
pp. 431-434 ◽  
Author(s):  
Danilo Catelani ◽  
Antonio Colombi
Keyword(s):  
Physicochemical Properties ◽  
Pseudomonas Putida ◽  
Dienoic Acid ◽  
Cleavage Product

The structure of 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid for the meta-cleavage product of 2,3-dihydroxybiphenyl by a Pseudomonas putida strain was demonstrated on the basis of its chemical and physicochemical properties and those of its derivatives.

scholarly journals HipH Catalyzes the Hydroxylation of 4-Hydroxyisophthalate to Protocatechuate in 2,4-Xylenol Catabolism by Pseudomonas putida NCIMB 9866

2015 ◽  
Vol 82 (2) ◽  
pp. 724-731 ◽  
Author(s):  
Hong-Jun Chao ◽  
Yan-Fei Chen ◽  
Ti Fang ◽  
Ying Xu ◽  
Wei E. Huang ◽  
...  
Keyword(s):  
Pseudomonas Putida ◽  
Environmental Protection Agency ◽  
High Performance ◽  
Specific Activity ◽  
Gene Knockout ◽  
Transcriptional Analysis ◽  
Ring Cleavage ◽  
Protection Agency ◽  
Content Type ◽  
Priority Pollutant

ABSTRACTIn addition to growing onp-cresol,Pseudomonas putidaNCIMB 9866 is the only reported strain capable of aerobically growing on 2,4-xylenol, which is listed as a priority pollutant by the U.S. Environmental Protection Agency. Several enzymes involved in the oxidation of thepara-methyl group, as well as the corresponding genes, have previously been reported. The enzyme catalyzing oxidation of the catabolic intermediate 4-hydroxyisophthalate to the ring cleavage substrate protocatechuate was also purified from strain NCIMB 9866, but its genetic determinant is still unavailable. In this study, the genehipH, encoding 4-hydroxyisophthalate hydroxylase, from strain NCIMB 9866 was cloned by transposon mutagenesis. Purified recombinant HipH-His6was found to be a dimer protein with a molecular mass of approximately 110 kDa. HipH-His6catalyzed the hydroxylation of 4-hydroxyisophthalate to protocatechuate with a specific activity of 1.54 U mg−1and showed apparentKmvalues of 11.40 ± 3.05 μM for 4-hydroxyisophthalate with NADPH and 11.23 ± 2.43 μM with NADH and similarKmvalues for NADPH and NADH (64.31 ± 13.16 and 72.76 ± 12.06 μM, respectively). The identity of protocatechuate generated from 4-hydroxyisophthalate hydroxylation by HipH-His6has also been confirmed by high-performance liquid chromatography and mass spectrometry. Gene transcriptional analysis, gene knockout, and complementation indicated thathipHis essential for 2,4-xylenol catabolism but not forp-cresol catabolism in this strain. This fills a gap in our understanding of the gene that encodes a critical step in 2,4-xylenol catabolism and also provides another example of biochemical and genetic diversity of microbial catabolism of structurally similar compounds.

scholarly journals Bacterial Degradation of N,N-Diethyl-m-Toluamide (DEET): Cloning and Heterologous Expression of DEET Hydrolase

2007 ◽  
Vol 73 (9) ◽  
pp. 3105-3108 ◽  
Author(s):  
Giomar Rivera-Cancel ◽  
Daniela Bocioaga ◽  
Anthony G. Hay
Keyword(s):  
Carbon Source ◽  
Heterologous Expression ◽  
Pseudomonas Putida ◽  
Sole Carbon Source ◽  
Bacterial Degradation ◽  
Ring Cleavage

ABSTRACT Pseudomonas putida DTB grew aerobically with N,N-diethyl-m-toluamide (DEET) as a sole carbon source, initially breaking it down into 3-methylbenzoate and diethylamine. The former was further metabolized via 3-methylcatechol and meta ring cleavage. A gene from DTB, dthA, was heterologously expressed and shown to encode the ability to hydrolyze DEET into 3-methylbenzoate and diethylamine.

scholarly journals p-Hydroxyphenylacetic Acid Metabolism inPseudomonas putida F6

2001 ◽  
Vol 183 (3) ◽  
pp. 928-933 ◽  
Author(s):  
Kevin E. O'Connor ◽  
Bernard Witholt ◽  
Wouter Duetz
Keyword(s):  
Pseudomonas Putida ◽  
Enzyme Activities ◽  
Acid Metabolism ◽  
Ring Cleavage ◽  
Acid Oxidation ◽  
Dihydroxyphenylacetic Acid ◽  
Cell Extracts ◽  
Hydroxyphenylacetic Acid ◽  
Brown Color

ABSTRACT Pseudomonas putida F6 was found to metabolizep-hydroxyphenylacetic acid through 3,4-dihydroxyphenylacetic acid, 3,4-dihydroxymandelic acid, and 3,4-dihydroxybenzaldehyde. Cell extracts of P. putida F6 catalyze the NAD(P)H-independent hydroxylation ofp-hydroxyphenylacetic acid to 3,4-dihydroxyphenylacetic acid which is further oxidized to 3,4-dihydroxymandelic acid. Oxidation and decarboxylation of the latter yields 3,4-dihydroxybenzaldehyde. A red-brown color accompanies all of the above enzyme activities and is probably due to the polymerization of quinone-like compounds. 3,4-Dihydroxybenzaldehyde is further metabolized through extradiol ring cleavage.

scholarly journals Oxidative side-chain and ring fission of pregnanes by Arthrobacter simplex

1988 ◽  
Vol 255 (3) ◽  
pp. 769-774 ◽  
Author(s):  
S B Mahato ◽  
S Banerjee ◽  
S Podder
Keyword(s):  
Methoxy Group ◽  
Ring Cleavage ◽  
Side Chain ◽  
Reaction Sequence ◽  
Arthrobacter Simplex ◽  
Ring Fission ◽  
Multistep Process ◽  
Microbial Preparation ◽  
17 Hydroxyprogesterone ◽  
Steroid Skeleton

Metabolic processes involving side-chain and ring cleavage of progesterone, 17-hydroxyprogesterone, 11-deoxycortisol and 16-dehydropregnenolone by Arthrobacter simplex were studied. The formation of the metabolites from progesterone indicates a pathway somewhat different from normal in the enzymic reaction sequence, and the 17-hydroxyprogesterone metabolites reveal a non-enzymic rearrangement step. The presence of a hydroxy group at C-21, as in 11-deoxycortisol, induces reduction of the C-20 carbonyl group. The microbial preparation of a novel androstane analogue, 17 beta-hydroxy-16 alpha-methoxyandrosta-1,4-dien-3-one, by incubation of 16-dehydropregnenolone with the bacterial strain was achieved. The formation of this metabolite is a multistep process involving a novel microbial generation of a methoxy group from a double-bond transformation in a steroid skeleton.

scholarly journals Cloning of a Sphingomonas paucimobilis SYK-6 Gene Encoding a Novel Oxygenase That Cleaves Lignin-Related Biphenyl and Characterization of the Enzyme

1998 ◽  
Vol 64 (7) ◽  
pp. 2520-2527 ◽  
Author(s):  
Xue Peng ◽  
Takashi Egashira ◽  
Kaoru Hanashiro ◽  
Eiji Masai ◽  
Seiji Nishikawa ◽  
...  
Keyword(s):  
Cosmid Library ◽  
Ring Cleavage ◽  
Sphingomonas Paucimobilis ◽  
Class Iii ◽  
Extradiol Dioxygenase ◽  
Gene Encoding ◽  
Reading Frame ◽  
Yellow Color ◽  
Ring Fission ◽  
Cleavage Enzyme

ABSTRACT Sphingomonas paucimobilis SYK-6 transforms 2,2′-dihydroxy-3,3′-dimethoxy-5,5′-dicarboxybiphenyl (DDVA), a lignin-related biphenyl compound, to 5-carboxyvanillic acid via 2,2′,3-trihydroxy-3′-methoxy-5,5′-dicarboxybiphenyl (OH-DDVA) as an intermediate (15). The ring fission of OH-DDVA is an essential step in the DDVA degradative pathway. A 15-kbEcoRI fragment isolated from the cosmid library complemented the growth deficiency of a mutant on OH-DDVA. Subcloning and deletion analysis showed that a 1.4-kb DNA fragment included the gene responsible for the ring fission of OH-DDVA. An open reading frame encoding 334 amino acids was identified and designatedligZ. The deduced amino acid sequence of LigZ had 18 to 21% identity with the class III extradiol dioxygenase family, including the β subunit (LigB) of protocatechuate 4,5-dioxygenase of SYK-6 (Y. Noda, S. Nishikawa, K.-I. Shiozuka, H. Kadokura, H. Nakajima, K. Yano, Y. Katayama, N. Morohoshi, T. Haraguchi, and M. Yamasaki, J. Bacteriol. 172:2704–2709, 1990), catechol 2,3-dioxygenase I (MpcI) ofAlcaligenes eutrophus JMP222 (M. Kabisch and P. Fortnagel, Nucleic Acids Res. 18:3405–3406, 1990), the catalytic subunit of themeta-cleavage enzyme (CarBb) for 2′-aminobiphenyl-2,3-diol from Pseudomonas sp. strain CA10 (S. I. Sato, N. Ouchiyama, T. Kimura, H. Nojiri, H. Yamane, and T. Omori, J. Bacteriol. 179:4841–4849, 1997), and 2,3-dihydroxyphenylpropionate 1,2-dioxygenase (MhpB) ofEscherichia coli (E. L. Spence, M. Kawamukai, J. Sanvoisin, H. Braven, and T. D. H. Bugg, J. Bacteriol. 178:5249–5256, 1996). The ring fission product formed from OH-DDVA by LigZ developed a yellow color with an absorption maximum at 455 nm, suggesting meta cleavage. Thus, LigZ was concluded to be a ring cleavage extradiol dioxygenase. LigZ activity was detected only for OH-DDVA and 2,2′,3,3′-tetrahydroxy-5,5′-dicarboxybiphenyl and was dependent on the ferrous ion.

scholarly journals Bacterial metabolism of 5-aminosalicylic acid. Initial ring cleavage

1992 ◽  
Vol 282 (3) ◽  
pp. 675-680 ◽  
Author(s):  
A Stolz ◽  
B Nörtemann ◽  
H J Knackmuss
Keyword(s):  
Bacterial Metabolism ◽  
Ring Cleavage ◽  
Pseudomonas Sp ◽  
Growth Substrate ◽  
Aminosalicylic Acid ◽  
Intact Cells ◽  
Cell Extracts ◽  
Ring Fission ◽  
Stoichiometric Reaction ◽  
Neutral Conditions

The metabolism of 5-aminosalicylate (5AS) by a bacterial strain, Pseudomonas sp. BN9, was studied. Intact cells of Pseudomonas sp. BN9 grown with 5AS oxidized 5AS and 2,5-dihydroxybenzoate (gentisate), whereas cells grown with gentisate oxidized only the growth substrate of all substituted salicylates tested. Cell extracts from Pseudomonas sp. BN9 catalysed the stoichiometric reaction of 1 mol of oxygen with 1 mol of 5AS to a metabolite with an intense u.v.-absorption maximum at 352 nm (pH 8.0). This metabolite was accumulated under neutral conditions, but was rapidly destroyed at acid pH. It was identified by m.s. and acid-catalysed deamination to fumarylpyruvate (trans-2,4-dioxohept-5-enedioic acid) as cis-4-amino-6-carboxy-2-oxohexa-3,5-dienoate, thus demonstrating direct cleavage of the monohydroxylated substrate 5AS to a non-aromatic ring-fission product. The enzyme responsible for conversion of 5AS was shown to be Fe(II)-dependent and to be distinct from gentisate 1,2-dioxygenase in strain BN9.

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