Q9KX40 (ESTB_BURGA) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
Esterase EstB
EC=3.1.1.-

Gene names

Name:

estB

Organism

Burkholderia gladioli (Pseudomonas marginata) (Phytomonas marginata)

Taxonomic identifier

28095 [NCBI]

Taxonomic lineage

cellular organisms › Bacteria › Proteobacteria › Betaproteobacteria › Burkholderiales › Burkholderiaceae › Burkholderia

Protein attributes

Sequence length	392 AA.
Sequence status	Complete.
Protein existence	Evidence 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 component	Cytoplasm
Molecular function	Hydrolase
Technical term	3D-structure
Gene Ontology (GO)
Cellular_component	cytoplasm Inferred from electronic annotation. Source: UniProtKB-SubCell
Molecular_function	hydrolase activity Inferred from electronic annotation. Source: UniProtKB-KW
Complete GO annotation...

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Chain

1 – 392

392

Esterase EstB

PRO_0000370699

Sites

Active site

75

1

Acyl-ester intermediate

Experimental info

Mutagenesis

75

1

S → A: Complete loss of esterase activity. Ref.1

Mutagenesis

149

1

S → A: Retains 30% esterase activity. Ref.1

Secondary structure

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392

Helix Strand Turn

Details...

Helix

16 – 31

16

Beta strand

36 – 44

9

Beta strand

47 – 58

12

Turn

59 – 62

4

Beta strand

70 – 72

3

Helix

74 – 76

3

Helix

77 – 90

14

Helix

100 – 102

3

Helix

121 – 125

5

Helix

134 – 137

4

Helix

142 – 146

5

Beta strand

151 – 153

3

Helix

159 – 168

10

Helix

184 – 196

13

Helix

200 – 207

8

Helix

209 – 212

4

Helix

224 – 226

3

Beta strand

235 – 237

3

Beta strand

245 – 248

4

Beta strand

255 – 259

5

Helix

261 – 265

5

Turn

273 – 275

3

Helix

281 – 293

13

Beta strand

295 – 298

4

Helix

300 – 307

8

Helix

313 – 315

3

Beta strand

322 – 324

3

Beta strand

326 – 332

7

Helix

334 – 337

4

Beta strand

346 – 350

5

Turn

351 – 353

3

Beta strand

354 – 359

6

Helix

360 – 362

3

Beta strand

364 – 373

10

Helix

375 – 378

4

Helix

380 – 389

10

Sequences

	Sequence		Length	Mass (Da)
	Q9KX40 [UniParc]. Last modified October 1, 2000. Version 1. Checksum: 60EC4764C3C499B7	FASTA	392	41,707
	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

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

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1CI8	X-ray	2.00	A/B	1-392	[»]
1CI9	X-ray	1.80	A/B	1-392	[»]

ProteinModelPortal

Q9KX40.

SMR

Q9KX40. Positions 15-391.

ModBase

Search...

Protein family/group databases

MEROPS

S12.950.

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Family and domain databases

Gene3D

3.40.710.10. 1 hit.

InterPro

IPR001466. Beta-lactam-related.
IPR012338. Beta-lactam/transpept-like.
IPR023650. Beta-lactam_class-A_AS.
[Graphical view]

Pfam

PF00144. Beta-lactamase. 1 hit.
[Graphical view]

SUPFAM

SSF56601. PBP_transp_fold. 1 hit.

PROSITE

PS00146. BETA_LACTAMASE_A. 1 hit.
[Graphical view]

ProtoNet

Search...

Other

EvolutionaryTrace

Q9KX40.

Entry information

Entry name

ESTB_BURGA

Accession

Primary (citable) accession number: Q9KX40

Entry history

Integrated into UniProtKB/Swiss-Prot:	May 5, 2009
Last sequence update:	October 1, 2000
Last modified:	April 3, 2013
This is version 49 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Prokaryotic Protein Annotation Program