(Below N is a link to NCBI taxonomic web page and E link to ESTHER at designed phylum.) > cellular organisms: NE > Bacteria: NE > Proteobacteria: NE > Betaproteobacteria: NE > Burkholderiales: NE > Comamonadaceae: NE > Delftia: NE > Delftia acidovorans: NE
LegendThis sequence has been compared to family alignement (MSA) red => minority aminoacid blue => majority aminoacid color intensity => conservation rate title => sequence position(MSA position)aminoacid rate Catalytic site Catalytic site in the MSA MNSRSLTKAIRFPTILALAGFSVLGACGGSDNDSSSNNQGAPAVAITVAG QVQAVDRLGMRRYFGIPFAAPPVGNLRWMPPAPPQSWAAPLAKTQSNAPC MQTGATDPLRLPNGTEDCLYLDVHAPATGEGPFPVMVWIHGGAFSIGGTI TYADPSPLVSKGVIVVNIAYRMGAMGFLGHPSLRAADGTVGNYGIMDQQA ALRWVQDNIAAFGGDKSNVTIFGESAGGFSVMTHLASPLSKGLFAKAIVQ SGGYGFDRQLTQAQLEAQSTSIVNSALAAAGVSCPTVDAACLRGLSAELV NNQLATAFTTANWSPVPSVDGKVLPKSIKATFVAGENNKVPLVNGSNQDE WSYFVASRELVAGPLTAAQYPSYLQTSLGLPPSLATVYPLTDYGTNTAQQ PSLAATAAGTDMHFSCPALNLSKRVLSQATPIFMYEFRDRTAIPSIGRNT ISFNQGAGHTYELQYLFNLRDLETAEHRDLQASMARYWTNFARTSNPNNG DPVATSWPAFTGPTKVLGLDVASAGGIRELATFETDHKCNTAWTSLTF
omamonas acidovorans strain TB-35 has an esterase that degrades solid polyester polyurethane (PUR). The structural gene, pudA, for the PUR esterase has now been cloned in Escherichia coli. When pudA was expressed in E. coli, the recombinant protein was able to degrade solid PUR. The predicted amino acid sequence contains the Gly-X1-Ser-X2-Gly motif characteristic of serine hydrolases. The highest degree of homology was detected with the Torpedo californica acetylcholinesterase (T AChE), possessing the Ser-His-Glu catalytic triad, with the glutamate residue replacing the usual aspartate residue. Similarity in the number and positions of cysteine and salt bonds was very apparent between PudA and T AChE, as were also identities of sequences and their positions in the alpha-helix and beta-strand regions between the two. In the neighborhood of the glutamate residue of the Ser199-His433-Glu324 catalytic domain of PudA, there were three hydrophobic domains, one of which constituted the surface-binding domain, which occurred in the C-terminus of most bacterial poly(hydroxyalkanoate)(PHA) depolymerases
Title: Determination of the polyester polyurethane breakdown products and distribution of the polyurethane degrading enzyme of Comamonas acidovorans strain TB-35 Nakajima-kambe T, Onuma F, Akutsu Y, Nakahara T Ref: J.Ferment.Bioeng, 83:456, 1997 : PubMed
Metabolites produced by the degradation of polyester polyurethane (PUR) by Comamonas acidovorans strain TB-35 were investigated. GC-MS analysis revealed that they were diethylene glycol, trimethylolpropane and adipic acid. These metabolites were considered to be derived from polyester segments of the PUR as a result of hydrolytic cleavage of ester bonds. In addition, when the culture broth was alkaline treated, a previously undetected product was detected by GC analysis. This product was identified as 2,4-diaminotoluene by GC-MS analysis. This indicates that an additional metabolite exists in the culture broth. This metabolite was considered to have been derived from polyisocyanate segments of the PUR. A preliminary study on a PUR degrading enzyme was also performed. Strain TB-35 produced two different esterases, one which is secreted to the culture broth and one which is bound to the cell surface. Between them, only cell-surface-bound esterase catalyzes the degradation of the polyester PUR.
Title: Isolation and characterization of a bacterium which utilizes polyester polyurethane as a sole carbon and nitrogen source Nakajima-kambe T, Onuma F, Kimpara N, Nakahara T Ref: FEMS Microbiology Letters, 129:39, 1995 : PubMed
Various soil samples were screened for the presence of microorganisms which have the ability to degrade polyurethane compounds. Two strains with good polyurethane degrading activity were isolated. The more active strain was tentatively identified as Comamonas acidovorans. This strain could utilize polyester-type polyurethanes but not the polyether-type polyurethanes as sole carbon and nitrogen sources. Adipic acid and diethylene glycol were probably the main degradation products when polyurethane was supplied as a sole carbon and nitrogen source. When ammonium nitrate was used as nitrogen source, only diethylene glycol was detected after growth on polyurethane.
The polyester-polyurethane (PUR)-degrading bacterium Comamonas acidovorans TB-35 produces two kinds of esterases, one cell-bound esterase (PUR esterase) and the other secreted in the culture broth (CBS esterase). In this study, the CBS esterase and the two recombinant esterases were purified. Identification of the physical and biochemical properties of the CBS and PUR esterases revealed that they have the same polypeptide from one gene. This finding was supported by the observation that Escherichia coli harboring the PUR esterase gene also produced two kinds of esterases. Though the PUR esterase degraded PUR and poly(diethylene glycol adipate), the soft segment of the PUR, the CBS esterase degraded only poly(diethylene glycol adipate). Furthermore, the hydrophobicity of the CBS esterase was lower than that of the PUR esterase. As the PUR esterase has been previously indicated to possess a PUR-binding domain, it was assumed that structural change around the PUR-binding domain of the CBS esterase was responsible for its inability to degrade PUR.
Title: Purification and Properties of a Polyester Polyurethane-Degrading Enzyme from Comamonas acidovorans TB-35 Akutsu Y, Nakajima-kambe T, Nomura N, Nakahara T Ref: Applied Environmental Microbiology, 64:62, 1998 : PubMed
A polyester polyurethane (PUR)-degrading enzyme, PUR esterase, derived from Comamonas acidovorans TB-35, a bacterium that utilizes polyester PUR as the sole carbon source, was purified until it showed a single band in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). This enzyme was bound to the cell surface and was extracted by addition of 0.2% N,N-bis(3-d-gluconamidopropyl)deoxycholamide (deoxy-BIGCHAP). The results of gel filtration and SDS-PAGE showed that the PUR esterase was a monomer with a molecular mass of about 62,000 Da. This enzyme, which is a kind of esterase, degraded solid polyester PUR, with diethylene glycol and adipic acid released as the degradation products. The optimum pH for this enzyme was 6.5, and the optimum temperature was 45 degrees C. PUR degradation by the PUR esterase was strongly inhibited by the addition of 0.04% deoxy-BIGCHAP. On the other hand, deoxy-BIGCHAP did not inhibit the activity when p-nitrophenyl acetate, a water-soluble compound, was used as a substrate. These observations indicated that this enzyme degrades PUR in a two-step reaction: hydrophobic adsorption to the PUR surface and hydrolysis of the ester bond of PUR.
omamonas acidovorans strain TB-35 has an esterase that degrades solid polyester polyurethane (PUR). The structural gene, pudA, for the PUR esterase has now been cloned in Escherichia coli. When pudA was expressed in E. coli, the recombinant protein was able to degrade solid PUR. The predicted amino acid sequence contains the Gly-X1-Ser-X2-Gly motif characteristic of serine hydrolases. The highest degree of homology was detected with the Torpedo californica acetylcholinesterase (T AChE), possessing the Ser-His-Glu catalytic triad, with the glutamate residue replacing the usual aspartate residue. Similarity in the number and positions of cysteine and salt bonds was very apparent between PudA and T AChE, as were also identities of sequences and their positions in the alpha-helix and beta-strand regions between the two. In the neighborhood of the glutamate residue of the Ser199-His433-Glu324 catalytic domain of PudA, there were three hydrophobic domains, one of which constituted the surface-binding domain, which occurred in the C-terminus of most bacterial poly(hydroxyalkanoate)(PHA) depolymerases
Title: Determination of the polyester polyurethane breakdown products and distribution of the polyurethane degrading enzyme of Comamonas acidovorans strain TB-35 Nakajima-kambe T, Onuma F, Akutsu Y, Nakahara T Ref: J.Ferment.Bioeng, 83:456, 1997 : PubMed
Metabolites produced by the degradation of polyester polyurethane (PUR) by Comamonas acidovorans strain TB-35 were investigated. GC-MS analysis revealed that they were diethylene glycol, trimethylolpropane and adipic acid. These metabolites were considered to be derived from polyester segments of the PUR as a result of hydrolytic cleavage of ester bonds. In addition, when the culture broth was alkaline treated, a previously undetected product was detected by GC analysis. This product was identified as 2,4-diaminotoluene by GC-MS analysis. This indicates that an additional metabolite exists in the culture broth. This metabolite was considered to have been derived from polyisocyanate segments of the PUR. A preliminary study on a PUR degrading enzyme was also performed. Strain TB-35 produced two different esterases, one which is secreted to the culture broth and one which is bound to the cell surface. Between them, only cell-surface-bound esterase catalyzes the degradation of the polyester PUR.
Title: Isolation and characterization of a bacterium which utilizes polyester polyurethane as a sole carbon and nitrogen source Nakajima-kambe T, Onuma F, Kimpara N, Nakahara T Ref: FEMS Microbiology Letters, 129:39, 1995 : PubMed
Various soil samples were screened for the presence of microorganisms which have the ability to degrade polyurethane compounds. Two strains with good polyurethane degrading activity were isolated. The more active strain was tentatively identified as Comamonas acidovorans. This strain could utilize polyester-type polyurethanes but not the polyether-type polyurethanes as sole carbon and nitrogen sources. Adipic acid and diethylene glycol were probably the main degradation products when polyurethane was supplied as a sole carbon and nitrogen source. When ammonium nitrate was used as nitrogen source, only diethylene glycol was detected after growth on polyurethane.
