PUL ID

PUL0607

PubMed

32520542, J Agric Food Chem. 2020 Jul 1;68(26):7032-7039. doi: 10.1021/acs.jafc.0c02404. Epub 2020 Jun 22.

Characterization method

enzymy activity assay, clone and expression, liquid chromatography and mass spectrometry

Genomic accession number

NZ_CP014224.1

Nucelotide position range

1677698-1717015

Substrate

porphyran

Loci

AXE80_RS06895-AXE80_RS07005

Species

Wenyingzhuangia fucanilytica strain CZ1127/1790137

Degradation or Biosynthesis

degradation

Gene Name

Locus Tag

Protein ID

Gene Position

GenBank Contig Range

EC Number

- AXE80_RS06895 WP_068825717.1 0 - 3228 (-) NZ_CP014224.1:1677698-1680926 -
- AXE80_RS06900 WP_068825719.1 3545 - 5690 (-) NZ_CP014224.1:1681243-1683388 -
- AXE80_RS14265 WP_083194605.1 5808 - 8859 (-) NZ_CP014224.1:1683506-1686557 -
- AXE80_RS06910 WP_068825721.1 9106 - 9877 (-) NZ_CP014224.1:1686804-1687575 -
- AXE80_RS06915 WP_068825723.1 10085 - 11663 (-) NZ_CP014224.1:1687783-1689361 -
- AXE80_RS06920 WP_068825725.1 11717 - 13577 (-) NZ_CP014224.1:1689415-1691275 -
- AXE80_RS06925 WP_068825727.1 13633 - 16762 (-) NZ_CP014224.1:1691331-1694460 -
- AXE80_RS06930 WP_068825729.1 17332 - 19027 (+) NZ_CP014224.1:1695030-1696725 -
- AXE80_RS06935 WP_083194606.1 19107 - 20634 (+) NZ_CP014224.1:1696805-1698332 -
- AXE80_RS06940 WP_068825731.1 20974 - 22156 (+) NZ_CP014224.1:1698672-1699854 -
- AXE80_RS06945 WP_068825732.1 22299 - 23550 (-) NZ_CP014224.1:1699997-1701248 -
- AXE80_RS06950 WP_068828752.1 23561 - 24452 (-) NZ_CP014224.1:1701259-1702150 -
- AXE80_RS06955 WP_068825734.1 24475 - 25348 (-) NZ_CP014224.1:1702173-1703046 -
- AXE80_RS06965 WP_068825739.1 25891 - 27424 (+) NZ_CP014224.1:1703589-1705122 -
- AXE80_RS06970 WP_068825740.1 27438 - 28449 (+) NZ_CP014224.1:1705136-1706147 -
- AXE80_RS06975 WP_068825742.1 28458 - 30111 (+) NZ_CP014224.1:1706156-1707809 -
- AXE80_RS06980 WP_068825744.1 30125 - 31577 (+) NZ_CP014224.1:1707823-1709275 -
- AXE80_RS06985 WP_068825746.1 31586 - 32414 (+) NZ_CP014224.1:1709284-1710112 -
- AXE80_RS06990 WP_068825748.1 32425 - 33217 (+) NZ_CP014224.1:1710123-1710915 -
fucP AXE80_RS06995 WP_068825750.1 33304 - 34639 (+) NZ_CP014224.1:1711002-1712337 -
- AXE80_RS07000 WP_068825752.1 34639 - 34981 (+) NZ_CP014224.1:1712337-1712679 -
- AXE80_RS07005 WP_068825754.1 35097 - 39318 (-) NZ_CP014224.1:1712795-1717016 -

Cluster number

1

Gene name

Gene position

Gene type

Found by CGCFinder?

- 1 - 3228 (-) CAZyme: GH2|CBM67|CBM51 Yes
- 3546 - 5690 (-) CAZyme: GH141|PL0 Yes
- 5809 - 8859 (-) CAZyme: GH105|GH154 Yes
- 9107 - 9877 (-) other Yes
- 10086 - 11663 (-) other Yes
- 11718 - 13577 (-) other Yes
- 13634 - 16762 (-) TC: gnl|TC-DB|Q45780|1.B.14.6.1 Yes
- 17333 - 19027 (+) other Yes
- 19108 - 20634 (+) other Yes
- 20975 - 22156 (+) CAZyme: GH16_11|GH16|3.2.1.178 Yes
- 22300 - 23550 (-) CAZyme: GH117 Yes
- 23562 - 24452 (-) CAZyme: GH16_11|GH16 Yes
- 24476 - 25348 (-) CAZyme: GH16_14 Yes
- 25892 - 27424 (+) CAZyme: GH29 Yes
- 27439 - 28449 (+) CDS No
- 28459 - 30111 (+) CDS No
- 30126 - 31577 (+) CDS No
- 31587 - 32414 (+) CDS No
- 32426 - 33217 (+) CDS No
- 33305 - 34639 (+) CDS No
- 34640 - 34981 (+) CDS No
- 35098 - 39318 (-) TF: DBD-Pfam|HTH_AraC,DBD-SUPERFAMILY|0035607,DBD-SUPERFAMILY|0035607 No

PUL ID

PUL0607

PubMed

32520542, J Agric Food Chem. 2020 Jul 1;68(26):7032-7039. doi: 10.1021/acs.jafc.0c02404. Epub 2020 Jun 22.

Title

Characterization of a Novel Porphyranase Accommodating Methyl-galactoses at Its Subsites.

Author

Zhang Y, Chang Y, Shen J, Mei X, Xue C

Abstract

Porphyran is the major polysaccharide of laver and mainly composed of 3-linked beta-d-galactopyranose (G) and 4-linked alpha-l-galactopyranose-6-sulfate (L6S) units. Structural heterogeneity of porphyran highly originates from the natural methylation on the O-6 position of G units (GMe). Here, a GH16 porphyranase Por16C_Wf was cloned from a porphyran-related polysaccharide utilization locus of Wenyingzhuangia fucanilytica and expressed in Escherichia coli. It hydrolyzed porphyran in a random endo-acting manner. Using a glycomics strategy combining liquid chromatography-mass spectrometry and glycoinformatics, the subsite specificity was clarified. Por16C_Wf accommodated both G and GMe at subsites -1 and +2. This is the first report on the sequence of porphyranases hydrolyzing consecutive methyl-porphyranobiose moieties, which shed light on the diversity in subsite specificity of porphyranases. Por16C_Wf was the first characterized enzyme in subfamily 14 of the GH16 family. The defined and novel activity of Por16C_Wf implied that it could serve as a favorable tool in the full degradation and structural investigation of porphyran.