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Cite: NAR/gkaa742 2020



PUL General Information

Literature Information

PUL ID

PUL0597

PubMed

25862223, Appl Environ Microbiol. 2015 Jun 15;81(12):4173-83. doi: 10.1128/AEM.00792-15. Epub 2015 Apr 10.

Characterization method

enzyme activity assay,enzyme kinetic analysis

Genomic accession number

KM066603

Nucelotide position range

110-6942

Substrate

arabinoxylan,arabino-xylooligosaccharide

Loci

xylHEFGD

Species

Corynebacterium alkanolyticum/909925

Degradation or Biosynthesis

degradation

Gene Name

Locus Tag

Protein ID

Gene Position

GenBank Contig Range

EC Number

xylH - AJY53614.1 109 - 1153 (-) KM066603.1:219-1263 -
xylE - AJY53615.1 1340 - 2619 (+) KM066603.1:1450-2729 -
xylF - AJY53616.1 2627 - 3620 (+) KM066603.1:2737-3730 -
xylG - AJY53617.1 3616 - 4510 (+) KM066603.1:3726-4620 -
xylD - AJY53618.1 4536 - 6942 (+) KM066603.1:4646-7052 -

Cluster number

1

Gene name

Gene position

Gene type

Found by CGCFinder?

- 110 - 1153 (-) TF: DBD-Pfam|LacI,DBD-SUPERFAMILY|0036955 No
- 1342 - 2619 (+) TC: gnl|TC-DB|Q9L027|3.A.1.1.21 Yes
- 2628 - 3620 (+) TC: gnl|TC-DB|Q72H67|3.A.1.1.25 Yes
- 3617 - 4510 (+) TC: gnl|TC-DB|G4FGN6|3.A.1.1.41 Yes
- 4537 - 6942 (+) CAZyme: GH3 Yes

PUL ID

PUL0597

PubMed

25862223, Appl Environ Microbiol. 2015 Jun 15;81(12):4173-83. doi: 10.1128/AEM.00792-15. Epub 2015 Apr 10.

Title

Functional Characterization of Corynebacterium alkanolyticum beta-Xylosidase and Xyloside ABC Transporter in Corynebacterium glutamicum.

Author

Watanabe A, Hiraga K, Suda M, Yukawa H, Inui M

Abstract

The Corynebacterium alkanolyticum xylEFGD gene cluster comprises the xylD gene that encodes an intracellular beta-xylosidase next to the xylEFG operon encoding a substrate-binding protein and two membrane permease proteins of a xyloside ABC transporter. Cloning of the cluster revealed a recombinant beta-xylosidase of moderately high activity (turnover for p-nitrophenyl-beta-d-xylopyranoside of 111 +/- 4 s(-1)), weak alpha-l-arabinofuranosidase activity (turnover for p-nitrophenyl-alpha-l-arabinofuranoside of 5 +/- 1 s(-1)), and high tolerance to product inhibition (Ki for xylose of 67.6 +/- 2.6 mM). Heterologous expression of the entire cluster under the control of the strong constitutive tac promoter in the Corynebacterium glutamicum xylose-fermenting strain X1 enabled the resultant strain X1EFGD to rapidly utilize not only xylooligosaccharides but also arabino-xylooligosaccharides. The ability to utilize arabino-xylooligosaccharides depended on cgR_2369, a gene encoding a multitask ATP-binding protein. Heterologous expression of the contiguous xylD gene in strain X1 led to strain X1D with 10-fold greater beta-xylosidase activity than strain X1EFGD, albeit with a total loss of arabino-xylooligosaccharide utilization ability and only half the ability to utilize xylooligosaccharides. The findings suggest some inherent ability of C. glutamicum to take up xylooligosaccharides, an ability that is enhanced by in the presence of a functional xylEFG-encoded xyloside ABC transporter. The finding that xylEFG imparts nonnative ability to take up arabino-xylooligosaccharides should be useful in constructing industrial strains with efficient fermentation of arabinoxylan, a major component of lignocellulosic biomass hydrolysates.