(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 > Gammaproteobacteria: NE > Pseudomonadales: NE > Pseudomonadaceae: NE > Pseudomonas: NE > Pseudomonas chlororaphis group: NE > Pseudomonas chlororaphis: 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 MGVFDYKNFTASDSKALFSDALAITLYSYHNIDNGFAEGYQHNGFGLGLP ATLVTALIGSGNSQGVIPGIPWNPDSEKAALDALHQAGWSTISAQQLGYD GKVDGRGTFFGEKAGYGTAQVEILGKYDAQGHLESIGIAFRGTSGPRESV ISDTIGDVINDLLAALGPKDYAKNYAGEAFGKLLGDVAAFAQANGLSGKD VLVSGHSLGGLGVNSLADLSSERWSGFYKDSNYIAYASPTQSASDKVLNI GYENDPVFRALDGSTFNLSSLGVHDAHQDSATNNIVNFNDHYASTLWNVL PSSILNIPTWLSHLPTGYGDGLSRVLESKFYDFTSKDSTIVVANLSDPAR ASTWVQDLNRNAETHKGSTFIIGSDGNDLIQGGSGNDYLEGRAGNDTFRD SGGYNIILGGQGSNTLDLQQSVKNYSFASDGAGTLYLRDANGGISMTRDI GAIQSKEPGFLWGLFKDDVIHQVTDQGLKAGGQLTQYASSVRGDAGDNVL KAHAGGDWLFGLDGNDHLIGGQGNDVFVGGAGNDLMEAGGGNNTFLFTGH FGQDRILGYQGGDKLVFMGVSGVLPDQDYRAHASSSGNDTVLTFGQDSVT LVGVSLEHLNGSGIVLA
References
1 moreTitle: Polyurethanases: Three-dimensional structures and molecular dynamics simulations of enzymes that degrade polyurethane do Canto VP, Thompson CE, Netz PA Ref: J Mol Graph Model, 89:82, 2019 : PubMed
The global production of plastics increases every year, because these materials are widely used in several segments of modern life. Polyurethanes are a very important class of polymers, used in many areas of everyday life, from automotive equipments to mattresses. The waste management usually involves accumulation in landfills, incineration, and reuse processes. However, bioremediation processes are being increasingly tested, due to the efficiency of enzymes in the degradation, besides adding value to the generated waste. Several experimental tests indicate that hydrolases, such as proteases, ureases, and esterases, are able to degrade polyurethanes. In this work, the three-dimensional structure of enzymes that are experimentally know to degrade polyurethanes were obtained for the first time, by the technique of homology modeling. The theoretical models showed good stereochemical quality and through molecular dynamics simulations analysis it was observed the stability of the structures. The molecular docking results indicated that all ligands, monomers of polyurethane, showed favorable interactions with the modeled enzymes.
Title: The polyester polyurethanase gene (pueA) from Pseudomonas chlororaphis encodes a lipase Stern RV, Howard GT Ref: FEMS Microbiology Letters, 185:163, 2000 : PubMed
A gene (pueA, polyurethane esterase A) encoding an extracellular polyurethanase (PueA) was cloned from Pseudomonas chlororaphis into Escherichia coli. The enzyme secreted from E. coli showed esterase activity when assayed with p-nitrophenyl acetate. Subcloning of a 3. 2-kb SalI-EcoRI fragment into a T7 RNA polymerase expression vector (pT7-6) produced a (35)S-labeled protein of 65 kDa. Nucleotide sequencing of pueA showed an open reading frame encoding a 65-kDa protein of 617 amino acid residues, with the serine hydrolase consensus sequence GXSXG. PueA was over-expressed using the pT7-6 vector transformed into E. coli BL21(DE3) and was purified in one step using Sephadex G-75.
Title: Purification and characterization of two polyurethanase enzymes from Pseudomonas chlororaphis Ruiz C, Main T, Hilliard NP, Howard GT Ref: International Biodeterioration & Biodegradation, 43:43, 1999 : PubMed
Two polyester polyurethane (PU)-degrading enzymes from Pseudomonas chlororaphis, a bacterium that utilizes polyester PU as the sole carbon and energy source,were purified to electrophoretic homogeneity as indicated by sodium dodecyl-polyacrylamide gel electrophoresis (SDS-PAGE). Both enzymes are extracellular, soluble proteins with molecular weight of 63,000 Da and 31,000 Da. The 63,000 Da protein exhibits both esterase and protease activities toward r-nitrophenylacetate and hide powder azure respectively. The enzyme has anoptimum pH of 8.5 for esterase activity and an optimum pH of 7.0 for protease activity. The 31,000 Da protein exhibits esterase activity toward r-nitrophenylacetate, butyrate and propionate,and has an optimum pH of 8.5. In addition, the enzyme activities of both proteins are heat stable after 10 min at 100 C and are inhibited 50% by the addition of 1 mM phenylmethylsulfonylfluoride indicating both are serine-hydrolases
1 lessTitle: Polyurethanases: Three-dimensional structures and molecular dynamics simulations of enzymes that degrade polyurethane do Canto VP, Thompson CE, Netz PA Ref: J Mol Graph Model, 89:82, 2019 : PubMed
The global production of plastics increases every year, because these materials are widely used in several segments of modern life. Polyurethanes are a very important class of polymers, used in many areas of everyday life, from automotive equipments to mattresses. The waste management usually involves accumulation in landfills, incineration, and reuse processes. However, bioremediation processes are being increasingly tested, due to the efficiency of enzymes in the degradation, besides adding value to the generated waste. Several experimental tests indicate that hydrolases, such as proteases, ureases, and esterases, are able to degrade polyurethanes. In this work, the three-dimensional structure of enzymes that are experimentally know to degrade polyurethanes were obtained for the first time, by the technique of homology modeling. The theoretical models showed good stereochemical quality and through molecular dynamics simulations analysis it was observed the stability of the structures. The molecular docking results indicated that all ligands, monomers of polyurethane, showed favorable interactions with the modeled enzymes.
AIMS: To better understand the role of PueA and PueB from Pseudomonas chlororaphis in polyurethane degradation, the present study was conducted to create insertional mutants in their respective genes. METHODS AND RESULTS: Growth kinetic studies showed that the pueA knockout mutant had a greater effect than the pueB knockout mutant. The pueA mutant had an 80% decrease in cell density from that of the wild type, while the pueB mutant had an 18% decrease in cell density. Polyurethane utilization followed Michaelis-Menten kinetics. The pueA and pueB mutants exhibited a 17% and 10% decrease respectively in growth rate using polyurethane when compared with the wild type. CONCLUSIONS: In this present study, pueA and pueB, are shown to be part of an ABC transporter gene cluster that consists of seven open reading frames. Mutational analysis results suggest that PueA may play a more major role in polyurethane degradation than PueB based on cell density and growth rates. SIGNIFICANCE AND IMPACT OF THE STUDY: The results from this study provide a starting point for the eventual enhancement and bioremediation of polyurethane waste. Understanding the role of polyurethane-degrading enzymes is useful for the creation of strains for this purpose.
Title: The polyester polyurethanase gene (pueA) from Pseudomonas chlororaphis encodes a lipase Stern RV, Howard GT Ref: FEMS Microbiology Letters, 185:163, 2000 : PubMed
A gene (pueA, polyurethane esterase A) encoding an extracellular polyurethanase (PueA) was cloned from Pseudomonas chlororaphis into Escherichia coli. The enzyme secreted from E. coli showed esterase activity when assayed with p-nitrophenyl acetate. Subcloning of a 3. 2-kb SalI-EcoRI fragment into a T7 RNA polymerase expression vector (pT7-6) produced a (35)S-labeled protein of 65 kDa. Nucleotide sequencing of pueA showed an open reading frame encoding a 65-kDa protein of 617 amino acid residues, with the serine hydrolase consensus sequence GXSXG. PueA was over-expressed using the pT7-6 vector transformed into E. coli BL21(DE3) and was purified in one step using Sephadex G-75.
Title: Purification and characterization of two polyurethanase enzymes from Pseudomonas chlororaphis Ruiz C, Main T, Hilliard NP, Howard GT Ref: International Biodeterioration & Biodegradation, 43:43, 1999 : PubMed
Two polyester polyurethane (PU)-degrading enzymes from Pseudomonas chlororaphis, a bacterium that utilizes polyester PU as the sole carbon and energy source,were purified to electrophoretic homogeneity as indicated by sodium dodecyl-polyacrylamide gel electrophoresis (SDS-PAGE). Both enzymes are extracellular, soluble proteins with molecular weight of 63,000 Da and 31,000 Da. The 63,000 Da protein exhibits both esterase and protease activities toward r-nitrophenylacetate and hide powder azure respectively. The enzyme has anoptimum pH of 8.5 for esterase activity and an optimum pH of 7.0 for protease activity. The 31,000 Da protein exhibits esterase activity toward r-nitrophenylacetate, butyrate and propionate,and has an optimum pH of 8.5. In addition, the enzyme activities of both proteins are heat stable after 10 min at 100 C and are inhibited 50% by the addition of 1 mM phenylmethylsulfonylfluoride indicating both are serine-hydrolases
