P0DX29 · PETH1_AMYMS
- ProteinCutinase
- Genecut1
- StatusUniProtKB reviewed (Swiss-Prot)
- Amino acids309 (go to sequence)
- Protein existenceEvidence at protein level
- Annotation score5/5
Function
function
Catalyzes the hydrolysis of cutin, a polyester that forms the structure of plant cuticle (By similarity).
Shows esterase activity towards p-nitrophenol-linked aliphatic esters (pNP-aliphatic esters) (PubMed:21145735, PubMed:33598102).
Has a preference for medium chain length (C-4 to C-12) fatty acid esters (PubMed:21145735, PubMed:33598102).
Active with p-nitrophenyl palmitate (p-NPP) as substrate (PubMed:19806375, PubMed:21145735, PubMed:35195792).
Hydrolyzes triacylglycerol substrates non-specifically with a preference for long, unsaturated fatty acyl chains with the highest activity for triolein (PubMed:21145735).
Substrates with cis-9 unsaturation are preferred over the saturated triacylglycerols (PubMed:21145735).
Hydrolyzes a wide range of natural oils, especially olive oil, with relatively high activity (PubMed:21145735).
Capable of catalyzing synthesis of the flavor ester isoamyl acetate by esterification of isoamyl alcohol using acetic acid as an acyl donor (PubMed:21145735).
Degrades synthetic aliphatic polyesters, namely poly(1,4-butylene succinate) extended with 1,6-diisocyanatohexane (PBSc-D) and poly(epsilon-caprolactone) (PCL) plastics (PubMed:33598102).
Does not degrade poly(lactic acid) (PLA) nor aromatic poly(ethylene terephthalate) (PET), the most abundant polyester plastic in the world (PubMed:33598102).
Shows esterase activity towards p-nitrophenol-linked aliphatic esters (pNP-aliphatic esters) (PubMed:21145735, PubMed:33598102).
Has a preference for medium chain length (C-4 to C-12) fatty acid esters (PubMed:21145735, PubMed:33598102).
Active with p-nitrophenyl palmitate (p-NPP) as substrate (PubMed:19806375, PubMed:21145735, PubMed:35195792).
Hydrolyzes triacylglycerol substrates non-specifically with a preference for long, unsaturated fatty acyl chains with the highest activity for triolein (PubMed:21145735).
Substrates with cis-9 unsaturation are preferred over the saturated triacylglycerols (PubMed:21145735).
Hydrolyzes a wide range of natural oils, especially olive oil, with relatively high activity (PubMed:21145735).
Capable of catalyzing synthesis of the flavor ester isoamyl acetate by esterification of isoamyl alcohol using acetic acid as an acyl donor (PubMed:21145735).
Degrades synthetic aliphatic polyesters, namely poly(1,4-butylene succinate) extended with 1,6-diisocyanatohexane (PBSc-D) and poly(epsilon-caprolactone) (PCL) plastics (PubMed:33598102).
Does not degrade poly(lactic acid) (PLA) nor aromatic poly(ethylene terephthalate) (PET), the most abundant polyester plastic in the world (PubMed:33598102).
Catalytic activity
- a carboxylic ester + H2O = a carboxylate + an alcohol + H+
- 1,2,3-tri-(9Z-octadecenoyl)-glycerol + H2O = (9Z)-octadecenoate + di-(9Z)-octadecenoylglycerol + H+
- (6-hydroxyhexanoyl)n + H2O = (6-hydroxyhexanoyl)n-1 + 6-hydroxyhexanoate + H+
Activity regulation
No effect on activity by SDS or chelating agents ethylenediaminetetraacetic acid (EDTA) or sodium citrate (PubMed:19806375, PubMed:21145735).
No effect on activity by metal ions Ag+, Ba2+, Ca2+, Co2+, Cu2+, Mn2+, Ni2+, Pb2+ or Zn2+ (PubMed:19806375, PubMed:21145735).
Activated by 1 mM digitonin and sodium deoxycholate, and reducing agents 1 mM 1,4-dithiothreitol, beta-mercaptoethanol and ascorbic acid (PubMed:21145735).
Activated by benzene, n-hexane, p-xylene and toluene (PubMed:19806375).
Activated by Fe3+ (PubMed:19806375, PubMed:21145735).
Inhibited slightly by 1 mM of different chain length fatty acids, and only marginally by 6.0 M urea (PubMed:21145735).
Inhibited strongly with chemical modification by reagents phenyl methyl sulfonylfluorid (PMSF), 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDAC), diethylpyrocarbonate (DEPC) and N-bromosuccinimide (NBS) (PubMed:21145735).
Inhibited by pyridine, DMSO, t-butanol and dodecane (PubMed:19806375).
Inhibited by Li+, Hg2+ and Mg2+ (PubMed:19806375, PubMed:21145735).
No inhibition with chemical modification by reagents N-acetylimidazole (NAI), citraconic anhydride (CA), iodoacetate (IA) and phenylglyoxal (PG) (PubMed:21145735).
No effect on activity by metal ions Ag+, Ba2+, Ca2+, Co2+, Cu2+, Mn2+, Ni2+, Pb2+ or Zn2+ (PubMed:19806375, PubMed:21145735).
Activated by 1 mM digitonin and sodium deoxycholate, and reducing agents 1 mM 1,4-dithiothreitol, beta-mercaptoethanol and ascorbic acid (PubMed:21145735).
Activated by benzene, n-hexane, p-xylene and toluene (PubMed:19806375).
Activated by Fe3+ (PubMed:19806375, PubMed:21145735).
Inhibited slightly by 1 mM of different chain length fatty acids, and only marginally by 6.0 M urea (PubMed:21145735).
Inhibited strongly with chemical modification by reagents phenyl methyl sulfonylfluorid (PMSF), 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDAC), diethylpyrocarbonate (DEPC) and N-bromosuccinimide (NBS) (PubMed:21145735).
Inhibited by pyridine, DMSO, t-butanol and dodecane (PubMed:19806375).
Inhibited by Li+, Hg2+ and Mg2+ (PubMed:19806375, PubMed:21145735).
No inhibition with chemical modification by reagents N-acetylimidazole (NAI), citraconic anhydride (CA), iodoacetate (IA) and phenylglyoxal (PG) (PubMed:21145735).
Biotechnology
May have potential in biocatalytic synthesis of esters by reverse hydrolysis, particularly in production of industrially important flavor and fragrance ester isoamyl acetate (PubMed:21145735).
After 72 hours of reaction, a yield of 34.4% and 16.2% of isoamyl acetate is achieved using Celite-immobilized and free enzyme, respectively (PubMed:21145735).
Due to high thermostability, organic solvent tolerance, activity in broad pH range, and specificity towards a broad substrate range, this protein may be a candidate for applications in non-aqueous biocatalytic processes including esterification of primary and secondary alcohols, random interesterification of different oils and fats in food industry, oil contaminated biodegradation and biodiesel production (PubMed:19806375, PubMed:21145735).
Has potential for application in bioremediation of environmental plastics (PubMed:33598102).
Recombinant protein expressed in E.coli may be tailored to degrade specific environmental polyester compounds (PubMed:35195792).
After 72 hours of reaction, a yield of 34.4% and 16.2% of isoamyl acetate is achieved using Celite-immobilized and free enzyme, respectively (PubMed:21145735).
Due to high thermostability, organic solvent tolerance, activity in broad pH range, and specificity towards a broad substrate range, this protein may be a candidate for applications in non-aqueous biocatalytic processes including esterification of primary and secondary alcohols, random interesterification of different oils and fats in food industry, oil contaminated biodegradation and biodiesel production (PubMed:19806375, PubMed:21145735).
Has potential for application in bioremediation of environmental plastics (PubMed:33598102).
Recombinant protein expressed in E.coli may be tailored to degrade specific environmental polyester compounds (PubMed:35195792).
Kinetics
KM | SUBSTRATE | pH | TEMPERATURE[C] | NOTES | EVIDENCE | |
---|---|---|---|---|---|---|
0.099 mM | p-nitrophenyl palmitate (p-NPP) | 8.0 | 60 |
Vmax | pH | TEMPERATURE[C] | NOTES | EVIDENCE | |
---|---|---|---|---|---|
2.53 mmol/min/mg | 8.0 | 60 | with p-nitrophenyl palmitate (p-NPP) as substrate |
kcat is 1468 sec-1 with p-nitrophenyl palmitate (p-NPP) as substrate (at pH 8.0 and 60 degrees Celsius).
pH Dependence
Optimum pH is 8.0 with p-nitrophenyl palmitate (p-NPP) as substrate (PubMed:19806375, PubMed:21145735).
At pH 7 and 9 displays around 90% of relative activity (PubMed:21145735).
Stable at pH range 6-9 retaining over 95% of relative activity after 24 hours (PubMed:21145735).
Stable even in acidic pH of 2-4 with more than 50% of residual activity after 24 hours (PubMed:19806375).
At pH 7 and 9 displays around 90% of relative activity (PubMed:21145735).
Stable at pH range 6-9 retaining over 95% of relative activity after 24 hours (PubMed:21145735).
Stable even in acidic pH of 2-4 with more than 50% of residual activity after 24 hours (PubMed:19806375).
Temperature Dependence
Optimum temperature is 60 degrees Celsius with p-nitrophenyl palmitate (p-NPP) as substrate (PubMed:19806375, PubMed:21145735).
Hydrolyzes p-NPP in the broad temperature range of 20-80 degrees Celsius (PubMed:19806375).
Highly thermostable (PubMed:19806375, PubMed:21145735).
Stable at 50-60 degrees Celsius retaining 90-100% of activity after 3 hours (PubMed:19806375, PubMed:21145735).
Has a half-life of over 30 minutes at 70 degrees Celsius (PubMed:19806375).
Hydrolyzes p-NPP in the broad temperature range of 20-80 degrees Celsius (PubMed:19806375).
Highly thermostable (PubMed:19806375, PubMed:21145735).
Stable at 50-60 degrees Celsius retaining 90-100% of activity after 3 hours (PubMed:19806375, PubMed:21145735).
Has a half-life of over 30 minutes at 70 degrees Celsius (PubMed:19806375).
Features
Showing features for active site.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Active site | 178 | Nucleophile | ||||
Sequence: S | ||||||
Active site | 224 | Charge relay system | ||||
Sequence: D | ||||||
Active site | 256 | Charge relay system | ||||
Sequence: H |
GO annotations
Aspect | Term | |
---|---|---|
Cellular Component | extracellular region | |
Molecular Function | carboxylesterase activity | |
Molecular Function | cutinase activity | |
Molecular Function | triglyceride lipase activity | |
Biological Process | medium-chain fatty acid catabolic process | |
Biological Process | triglyceride catabolic process |
Keywords
- Molecular function
- Biological process
Enzyme and pathway databases
Names & Taxonomy
Protein names
- Recommended nameCutinase
- EC number
- Alternative names
Gene names
Organism names
- Strains
- Taxonomic lineageBacteria > Actinomycetota > Actinomycetes > Pseudonocardiales > Pseudonocardiaceae > Amycolatopsis
Accessions
- Primary accessionP0DX29
- Secondary accessions
Proteomes
Subcellular Location
Phenotypes & Variants
PTM/Processing
Features
Showing features for signal, chain, disulfide bond.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Signal | 1-47 | |||||
Sequence: MSALTSQPTSSGSSEKIPRLRGWRAKAAGVVLAALALTTGVAAPAPA | ||||||
Chain | PRO_5003399567 | 48-309 | Cutinase | |||
Sequence: AANPYERGPDPTTASIEATSGSFATSTVTVSRLAVSGFGGGTIYYPTTTTAGTFGALSIAPGFTATQSSIAWLGPRLASQGFVVFTIDTLTTSDQPDSRGRQLLASLDYLTQQSSVRSRIDSTRLGVVGHSMGGGGTLEAARSRPTLQAAVPLTAWDLTKNWSTLQVPTLVVGAQSDTVAPVASHSIPFYTSLPSTLDRAYLELRGASHFAPNSPNTTIAKYTLSWLKRFIDNDTRYEQFLCPIPSTSLSISDYRGNCPHNG | ||||||
Disulfide bond | 289↔305 | |||||
Sequence: CPIPSTSLSISDYRGNC |
Keywords
- PTM
Sequence
- Sequence statusComplete
- Sequence processingThe displayed sequence is further processed into a mature form.
- Length309
- Mass (Da)32,377
- Last updated2023-09-13 v1
- Checksum67095AC3AE1A453E
Features
Showing features for sequence conflict.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Sequence conflict | 67 | in Ref. 3; AA sequence | ||||
Sequence: S → R |
Keywords
- Technical term
Sequence databases
Nucleotide Sequence | Protein Sequence | Molecule Type | Status | |
---|---|---|---|---|
CP002896 EMBL· GenBank· DDBJ | AEK46168.1 EMBL· GenBank· DDBJ | Genomic DNA |