Reviewed, UniProtKB/Swiss-Prot P56216 (VAOX_PENSI)

Names and origin

Protein names

Recommended name:
    Vanillyl-alcohol oxidase
    EC=1.1.3.38
Alternative name(s):
    Aryl-alcohol oxidase
    4-allylphenol oxidase

Gene names

Name:

VAOA

Organism

Penicillium simplicissimum

Taxonomic identifier

69488 [NCBI]

Taxonomic lineage

Eukaryota › Fungi › Dikarya › Ascomycota › Pezizomycotina › Eurotiomycetes › Eurotiomycetidae › Eurotiales › Trichocomaceae › mitosporic Trichocomaceae › Penicillium

Protein attributes

Sequence length	560 AA.
Sequence status	Complete.
Sequence processing	The displayed sequence is not processed.
Protein existence	Evidence at protein level.

General annotation (Comments)

Function	Catalyzes the conversion of vanillin alcohol to vanillin, and also the conversion of a wide range of phenolic compounds bearing side chains of variable size at the para position of the aromatic ring. Crucial for the degradation of the secondary metabolites derived from the degradation of the lignin. Catalyzes besides the oxidation of 4-hydroxybenzyl alcohols, the oxidative deamination of 4-hydroxybenzylamines, the oxidative demethylation of 4-(methoxy-methyl)phenols and the oxidative hydration of 4-allylphenols. Most active with 4-allylphenols.
Catalytic activity	Vanillyl alcohol + O2 = vanillin + H2O2.
Cofactor	Binds 1 FAD covalently per subunit.
Enzyme regulation	Competitively inhibited by cinnamyl and coniferyl alcohols and by isoeugenol.
Subunit structure	Homooctamer (tetramer of tightly interacting dimers).
Subcellular location	Peroxisome. Cytoplasm. Ref.7
Induction	By 4-methoxybenzyl alcohols, anisyl and veratryl alcohols. Repressed by carbon catabolite.
Sequence similarities	Contains 1 FAD-binding PCMH-type domain. To bacterial flavocytochrome p-cresol methyl hydroxylase.
Biophysicochemical properties	pH dependence: Optimum pH is about 10. Temperature dependence: Optimum temperature is 38 degrees Celsius.

Ontologies

Keywords
Biological process	Methanol utilization
Cellular component	Cytoplasm Peroxisome
Ligand	FAD Flavoprotein
Molecular function	Oxidoreductase
Technical term	3D-structure Direct protein sequencing
Gene Ontology (GO)
Biological process	methanol metabolic process Inferred from electronic annotation. Source: UniProtKB-KW oxidation reduction Inferred from electronic annotation. Source: UniProtKB-KW
Cellular component	peroxisome Inferred from electronic annotation. Source: UniProtKB-SubCell
Molecular function	FAD binding Inferred from electronic annotation. Source: InterPro vanillyl-alcohol oxidase activity Inferred from electronic annotation. Source: EC
Complete GO annotation...

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Chain

1 – 560

560

Vanillyl-alcohol oxidase

PRO_0000065763

Regions

Domain

67 – 272

206

FAD-binding PCMH-type

Sites

Active site

108

1

Active site

503

1

Active site

504

1

Site

170

1

Important for the catalytic mechanism; Involved in substrate deprotonation

Amino acid modifications

Modified residue

422

1

Tele-8alpha-FAD histidine

Experimental info

Sequence conflict

274

1

R → G in CAA75722. Ref.1

Sequence conflict

330

1

R → K in CAA75722. Ref.1

Secondary structure

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560

Helix Strand Turn

Details...

Helix

18 – 32

15

Helix

34 – 36

3

Beta strand

37 – 39

3

Beta strand

51 – 54

4

Beta strand

56 – 58

3

Beta strand

72 – 75

4

Helix

80 – 93

14

Beta strand

97 – 102

6

Turn

106 – 111

6

Beta strand

119 – 122

4

Turn

124 – 126

3

Beta strand

130 – 134

5

Turn

135 – 138

4

Beta strand

139 – 142

4

Helix

148 – 157

10

Turn

161 – 163

3

Helix

176 – 181

6

Helix

194 – 196

3

Beta strand

197 – 204

8

Beta strand

209 – 211

3

Helix

213 – 216

4

Helix

223 – 225

3

Helix

230 – 232

3

Turn

237 – 241

5

Helix

251 – 254

4

Beta strand

255 – 268

14

Beta strand

276 – 283

8

Helix

286 – 288

3

Helix

289 – 301

13

Beta strand

310 – 313

4

Helix

314 – 321

8

Helix

324 – 326

3

Helix

336 – 346

11

Beta strand

350 – 359

10

Helix

361 – 375

15

Beta strand

382 – 384

3

Helix

386 – 388

3

Helix

394 – 401

8

Turn

402 – 404

3

Helix

409 – 416

8

Beta strand

420 – 425

6

Beta strand

427 – 429

3

Helix

433 – 450

18

Beta strand

456 – 460

5

Beta strand

465 – 474

10

Helix

478 – 497

20

Helix

507 – 509

3

Helix

510 – 516

7

Helix

519 – 535

17

Helix

545 – 547

3

Helix

555 – 558

4

Sequences

	Sequence		Length	Mass (Da)
	P56216-1 [UniParc]. Last modified July 15, 1998. Version 1. Checksum: C3B4E4A966C1BCEE	FASTA	560	63,035
	10 20 30 40 50 60 MSKTQEFRPL TLPPKLSLSD FNEFIQDIIR IVGSENVEVI SSKDQIVDGS YMKPTHTHDP 70 80 90 100 110 120 HHVMDQDYFL ASAIVAPRNV ADVQSIVGLA NKFSFPLWPI SIGRNSGYGG AAPRVSGSVV 130 140 150 160 170 180 LDMGKNMNRV LEVNVEGAYC VVEPGVTYHD LHNYLEANNL RDKLWLDVPD LGGGSVLGNA 190 200 210 220 230 240 VERGVGYTPY GDHWMMHSGM EVVLANGELL RTGMGALPDP KRPETMGLKP EDQPWSKIAH 250 260 270 280 290 300 LFPYGFGPYI DGLFSQSNMG IVTKIGIWLM PNPRGYQSYL ITLPKDGDLK QAVDIIRPLR 310 320 330 340 350 360 LGMALQNVPT IRHILLDAAV LGDKRSYSSR TEPLSDEELD KIAKQLNLGR WNFYGALYGP 370 380 390 400 410 420 EPIRRVLWET IKDAFSAIPG VKFYFPEDTP ENSVLRVRDK TMQGIPTYDE LKWIDWLPNG 430 440 450 460 470 480 AHLFFSPIAK VSGEDAMMQY AVTKKRCQEA GLDFIGTFTV GMREMHHIVC IVFNKKDLIQ 490 500 510 520 530 540 KRKVQWLMRT LIDDCAANGW GEYRTHLAFM DQIMETYNWN NSSFLRFNEV LKNAVDPNGI 550 560 IAPGKSGVWP SQYSHVTWKL

References

[1]	"Molecular cloning, sequencing, and heterologous expression of the vaoA gene from Penicillium simplicissimum CBS 170.90 encoding vanillyl-alcohol oxidase." Benen J.A.E., Sanchez-Torres P., Wagemaker M.J.M., Fraaije M.W., van Berkel W.J.H., Visser J. J. Biol. Chem. 273:7865-7872(1998) [PubMed: 9525880] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 6-30 AND 130-148. Strain: ATCC 90172 / CBS 170.90 / PAZ 1.
[2]	"Crystallization and preliminary X-ray analysis of the flavoenzyme vanillyl-alcohol oxidase from Penicillium simplicissimum." Mattevi A., Fraaije M.W., Coda A., van Berkel W.J.H. Proteins 27:601-603(1997) [PubMed: 9141139] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.5 ANGSTROMS). Strain: ATCC 90172 / CBS 170.90 / PAZ 1.
[3]	"Covalent flavinylation is essential for efficient redox catalysis in vanillyl-alcohol oxidase." Fraaije M.W., van den Heuvel R.H.H., van Berkel W.J.H., Mattevi A. J. Biol. Chem. 274:35514-35520(1999) [PubMed: 10585424] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS) OF MUTANTS. Strain: ATCC 90172 / CBS 170.90 / PAZ 1.
[4]	"Asp-170 is crucial for the redox properties of vanillyl-alcohol oxidase." van den Heuvel R.H.H., Fraaije M.W., Mattevi A., van Berkel W.J.H. J. Biol. Chem. 275:14799-14808(2000) [PubMed: 10809721] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.8 ANGSTROMS) OF D170S MUTANT.
[5]	"Inversion of stereospecificity of vanillyl-alcohol oxidase." van den Heuvel R.H.H., Fraaije M.W., Ferrer M., Mattevi A., van Berkel W.J.H. Proc. Natl. Acad. Sci. U.S.A. 97:9455-9460(2000) [PubMed: 10920192] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.75 ANGSTROMS) OF D170S/T457E MUTANT.
[6]	"Structural analysis of flavinylation in vanillyl-alcohol oxidase." Fraaije M.W., van den Heuvel R.H.H., van Berkel W.J.H., Mattevi A. J. Biol. Chem. 275:38654-38658(2000) [PubMed: 10984479] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.1 ANGSTROMS) OF MUTANTS. Strain: ATCC 90172 / CBS 170.90 / PAZ 1.
[7]	"Subcellular localization of vanillyl-alcohol oxidase in Penicillium simplicissimum." Fraaije M.W., Sjollema K.A., Veenhuis M., van Berkel W.J.H. FEBS Lett. 422:65-68(1998) [PubMed: 9475171] [Abstract] Cited for: SUBCELLULAR LOCATION.
[8]	"Purification and characterization of vanillyl-alcohol oxidase from Penicillium simplicissimum. A novel aromatic alcohol oxidase containing covalently bound FAD." de Jong E., van Berkel W.J.H., van der Zwan R.P., de Bont J.A.M. Eur. J. Biochem. 208:651-657(1992) [PubMed: 1396672] [Abstract] Cited for: CHARACTERIZATION. Strain: ATCC 90172 / CBS 170.90 / PAZ 1.
[9]	"Substrate specificity of flavin-dependent vanillyl-alcohol oxidase from Penicillium simplicissimum. Evidence for the production of 4-hydroxycinnamyl alcohols from 4-allylphenols." Fraaije M.W., Veeger C., van Berkel W.J.H. Eur. J. Biochem. 234:271-277(1995) [PubMed: 8529652] [Abstract] Cited for: SUBSTRATE SPECIFICITY. Strain: ATCC 90172 / CBS 170.90 / PAZ 1.
+	Additional computationally mapped references.

Cross-references

Sequence databases

EMBLGenBankDDBJ

Y15627 Genomic DNA. Translation: CAA75722.1.

3D structure databases

PDBPDBe

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1AHU	X-ray	2.70	A/B	1-560	[»]
1AHV	X-ray	3.10	A/B	1-560	[»]
1AHZ	X-ray	3.30	A/B	1-560	[»]
1DZN	X-ray	2.80	A/B	1-560	[»]
1E0Y	X-ray	2.75	A/B	1-560	[»]
1E8F	X-ray	2.90	A/B	1-560	[»]
1E8G	X-ray	2.10	A/B	1-560	[»]
1E8H	X-ray	2.60	A/B	1-560	[»]
1QLT	X-ray	2.20	A/B	1-560	[»]
1QLU	X-ray	2.40	A/B	1-560	[»]
1VAO	X-ray	2.50	A/B	1-560	[»]
1W1J	X-ray	2.70	A/B	1-560	[»]
1W1K	X-ray	2.55	A/B	1-560	[»]
1W1L	X-ray	2.70	A/B	1-560	[»]
1W1M	X-ray	3.00	A/B	1-560	[»]
2VAO	X-ray	2.80	A/B	1-560	[»]

ModBase

Search...

Enzyme and pathway databases

BRENDA

1.1.3.38. 6842.

Family and domain databases

InterPro

IPR016166. FAD-bd_2.
IPR016167. FAD-bd_2_sub1.
IPR016168. FAD-linked_Oxase_FAD-bd_sub2.
IPR004113. FAD-linked_oxidase_C.
IPR006094. Oxid_FAD_bind_N.
IPR016170. Vanillyl_alc_oxidase_C-sub1.
IPR016171. Vanillyl_alc_oxidase_C-sub2.
[Graphical view]

Gene3D

G3DSA:3.30.43.10. FAD-binding_2_sub1. 1 hit.
G3DSA:3.30.465.20. FAD-linked_oxidase_FAD-bd_sub2. 1 hit.
G3DSA:3.40.462.10. Vanillyl_alc_oxidase_C-sub1. 1 hit.
G3DSA:1.10.45.10. Vanillyl_alc_oxidase_C-sub2. 1 hit.

Pfam

PF02913. FAD-oxidase_C. 1 hit.
PF01565. FAD_binding_4. 1 hit.
[Graphical view]

PROSITE

PS51387. FAD_PCMH. 1 hit.
[Graphical view]

ProtoNet

Search...

Entry information

Entry name

VAOX_PENSI

Accession

Primary (citable) accession number: P56216
Secondary accession number(s): O60049

Entry history

Integrated into UniProtKB/Swiss-Prot:	July 15, 1998
Last sequence update:	July 15, 1998
Last modified:	June 16, 2009
This is version 67 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation project

FPAP (Fungal Proteome Annotation Project)