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Q9KX40 (ESTB_BURGA) Reviewed, UniProtKB/Swiss-Prot

Last modified April 16, 2014. Version 52. Feed History...

Clusters with 100%, 90%, 50% identity | Documents (2) | Third-party data text xml rdf/xml gff fasta
to top of pageNames·Attributes·General annotation·Ontologies·Sequence annotation·Sequences·References·Cross-refs·Entry info·DocumentsCustomize order

Names and origin

Protein namesRecommended name:
Esterase EstB

EC=3.1.1.-
Gene names
Name:estB
OrganismBurkholderia gladioli (Pseudomonas marginata) (Phytomonas marginata)
Taxonomic identifier28095 [NCBI]
Taxonomic lineageBacteriaProteobacteriaBetaproteobacteriaBurkholderialesBurkholderiaceaeBurkholderia

Protein attributes

Sequence length392 AA.
Sequence statusComplete.
Protein existenceEvidence at protein level

General annotation (Comments)

Function

Acts on short-chain (C4-C6) fatty acid esters and triglycerides, including tertiary alcohol esters. Activity on p-nitrophenyl esters is generally higher than on o-nitrophenyl esters. Lacks beta-lactamase activity; it hydrolyzes the ester bond of cephalosporin substrates but there is no opening of the beta-lactam ring observed.

Enzyme regulation

Strongly inhibited by eserin, NaF, HgCl2, SDS and Triton X-100.

Subcellular location

Cytoplasm Probable Ref.1.

Biotechnological use

Directed evolutionary mutagenesis has created an enzyme with 40-fold higher stability in organic solvents. It contains the following changes: Leu-8, Ser-132, Arg-134, Cys-155, Gly-251, Val-311 and Lys-316 and could have applications in antibiotic synthesis (Ref.2).

Sequence similarities

Belongs to the class-A beta-lactamase family.

Biophysicochemical properties

Kinetic parameters:

Esterase, not beta-lactamase activity of the enzyme.

KM=1.3 mM for cephalosporin C

KM=1.0 mM for 7-amino cephalosporinic acid

Vmax=79 µmol/min/mg enzyme for cephalosporin C

Vmax=77 µmol/min/mg enzyme for 7-amino cephalosporinic acid

pH dependence:

Optimum pH is 7.0.

Temperature dependence:

Optimum temperature is 43 degrees Celsius.

Ontologies

Keywords
   Cellular componentCytoplasm
   Molecular functionHydrolase
   Technical term3D-structure
Gene Ontology (GO)
   Cellular_componentcytoplasm

Inferred from electronic annotation. Source: UniProtKB-SubCell

   Molecular_functionhydrolase activity

Inferred from electronic annotation. Source: UniProtKB-KW

Complete GO annotation...

Sequence annotation (Features)

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifier

Molecule processing

Chain1 – 392392Esterase EstB
PRO_0000370699

Sites

Active site751Acyl-ester intermediate

Experimental info

Mutagenesis751S → A: Complete loss of esterase activity. Ref.1
Mutagenesis1491S → A: Retains 30% esterase activity. Ref.1

Secondary structure

.................................................................... 392
Helix Strand Turn

Details...

Sequences

Sequence LengthMass (Da)Tools
Q9KX40 [UniParc].

Last modified October 1, 2000. Version 1.
Checksum: 60EC4764C3C499B7

FASTA39241,707
        10         20         30         40         50         60 
MTAASLDPTA FSLDAASLAA RLDAVFDQAL RERRLVGAVA IVARHGEILY RRAQGLADRE 

        70         80         90        100        110        120 
AGRPMREDTL FRLASVTKPI VALAVLRLVA RGELALDAPV TRWLPEFRPR LADGSEPLVT 

       130        140        150        160        170        180 
IHHLLTHTSG LGYWLLEGAG SVYDRLGISD GIDLRDFDLD ENLRRLASAP LSFAPGSGWQ 

       190        200        210        220        230        240 
YSLALDVLGA VVERATGQPL AAAVDALVAQ PLGMRDCGFV SAEPERFAVP YHDGQPEPVR 

       250        260        270        280        290        300 
MRDGIEVPLP EGHGAAVRFA PSRVFEPGAY PSGGAGMYGS ADDVLRALEA IRANPGFLPE 

       310        320        330        340        350        360 
TLADAARRDQ AGVGAETRGP GWGFGYLSAV LDDPAAAGTP QHAGTLQWGG VYGHSWFVDR 

       370        380        390 
ALGLSVLLLT NTAYEGMSGP LTIALRDAVY AR 

« Hide

References

[1]"A novel esterase from Burkholderia gladioli which shows high deacetylation activity on cephalosporins is related to beta-lactamases and DD-peptidases."
Petersen E.I., Valinger G., Soelkner B., Stubenrauch G., Schwab H.
J. Biotechnol. 89:11-25(2001) [PubMed] [Europe PMC] [Abstract]
Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], CHARACTERIZATION AS AN ESTERASE, MUTAGENESIS OF SER-75 AND SER-149, ABSENCE OF BETA-LACTAMASE ACTIVITY, PROBABLE SUBCELLULAR LOCATION.
Strain: ATCC 10248 / NCPPB 1891.
[2]"Stability and activity improvement of cephalosporin esterase EstB from Burkholderia gladioli by directed evolution and structural interpretation of muteins."
Valinger G., Hermann M., Wagner U.G., Schwab H.
J. Biotechnol. 129:98-108(2007) [PubMed] [Europe PMC] [Abstract]
Cited for: RANDOM MUTAGENESIS TO CREATE MORE STABLE ORGANIC SOLVENT-RESISTANT ENZYME.
Strain: ATCC 10248 / NCPPB 1891.
[3]"Inverting enantioselectivity of Burkholderia gladioli esterase EstB by directed and designed evolution."
Ivancic M., Valinger G., Gruber K., Schwab H.
J. Biotechnol. 129:109-122(2007) [PubMed] [Europe PMC] [Abstract]
Cited for: RANDOM MUTAGENESIS TO ALTER ENANTIOSELECTIVITY.
Strain: ATCC 10248 / NCPPB 1891.
[4]"EstB from Burkholderia gladioli: a novel esterase with a beta-lactamase fold reveals steric factors to discriminate between esterolytic and beta-lactam cleaving activity."
Wagner U.G., Petersen E.I., Schwab H., Kratky C.
Protein Sci. 11:467-478(2002) [PubMed] [Europe PMC] [Abstract]
Cited for: X-RAY CRYSTALLOGRAPHY (1.8 ANGSTROMS) WITH AND WITHOUT THE INHIBITOR DIISOPROPYL-FLUOROPHOSPHATE.
Strain: ATCC 10248 / NCPPB 1891.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ
U33634 Genomic DNA. Translation: AAF59826.1.

3D structure databases

PDBe
RCSB-PDB
PDBj
EntryMethodResolution (Å)ChainPositionsPDBsum
1CI8X-ray2.00A/B1-392[»]
1CI9X-ray1.80A/B1-392[»]
ProteinModelPortalQ9KX40.
SMRQ9KX40. Positions 15-391.
ModBaseSearch...
MobiDBSearch...

Protein family/group databases

MEROPSS12.950.

Protocols and materials databases

StructuralBiologyKnowledgebaseSearch...

Family and domain databases

Gene3D3.40.710.10. 1 hit.
InterProIPR001466. Beta-lactam-related.
IPR012338. Beta-lactam/transpept-like.
IPR023650. Beta-lactam_class-A_AS.
[Graphical view]
PfamPF00144. Beta-lactamase. 1 hit.
[Graphical view]
SUPFAMSSF56601. SSF56601. 1 hit.
PROSITEPS00146. BETA_LACTAMASE_A. 1 hit.
[Graphical view]
ProtoNetSearch...

Other

EvolutionaryTraceQ9KX40.

Entry information

Entry nameESTB_BURGA
AccessionPrimary (citable) accession number: Q9KX40
Entry history
Integrated into UniProtKB/Swiss-Prot: May 5, 2009
Last sequence update: October 1, 2000
Last modified: April 16, 2014
This is version 52 of the entry and version 1 of the sequence. [Complete history]
Entry statusReviewed (UniProtKB/Swiss-Prot)
Annotation programProkaryotic Protein Annotation Program

Relevant documents

SIMILARITY comments

Index of protein domains and families

PDB cross-references

Index of Protein Data Bank (PDB) cross-references