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Cite: NAR/gkaa742 2020



PUL General Information

Literature Information

PUL ID

PUL0390

PubMed

21255309, Microb Biotechnol. 2010 Jan;3(1):84-92. doi: 10.1111/j.1751-7915.2009.00150.x. Epub 2009 Sep 18.

Characterization method

enzyme activity assay

Genomic accession number

NC_000853.1

Nucelotide position range

51632-81458

Substrate

acetylated glucuronoxylan

Loci

TM0055-TM0077

Species

Thermotoga maritima/2336

Degradation or Biosynthesis

degradation

Gene Name

Locus Tag

Protein ID

Gene Position

GenBank Contig Range

EC Number

- TM_RS00250 WP_004082543.1 0 - 2025 (-) NC_000853.1:51632-53657 3.2.1.139
- TM_RS00255 WP_004082544.1 2081 - 4055 (-) NC_000853.1:53713-55687 -
- TM_RS00260 WP_004082545.1 4068 - 5085 (-) NC_000853.1:55700-56717 -
- TM_RS00265 WP_004082546.1 5081 - 6077 (-) NC_000853.1:56713-57709 -
- TM_RS00270 WP_004082547.1 6090 - 7179 (-) NC_000853.1:57722-58811 -
- TM_RS00275 WP_004082548.1 7207 - 8194 (-) NC_000853.1:58839-59826 -
- TM_RS00280 WP_004082550.1 8512 - 11692 (+) NC_000853.1:60144-63324 3.2.1.8
- TM_RS00285 WP_004082552.1 11747 - 12287 (+) NC_000853.1:63379-63919 -
- TM_RS00290 WP_004082554.1 12333 - 12780 (-) NC_000853.1:63965-64412 -
uxaC TM_RS00295 WP_004082556.1 12791 - 14147 (-) NC_000853.1:64423-65779 5.3.1.12
- TM_RS00300 WP_004082558.1 14248 - 14989 (+) NC_000853.1:65880-66621 -
- TM_RS00305 WP_004082560.1 15057 - 15669 (+) NC_000853.1:66689-67301 -
- TM_RS00310 WP_004082562.1 15665 - 16685 (+) NC_000853.1:67297-68317 -
uxuB TM_RS00315 WP_004082564.1 16697 - 18317 (+) NC_000853.1:68329-69949 1.1.1.57
uxuA TM_RS00320 WP_004082566.1 18313 - 19396 (+) NC_000853.1:69945-71028 4.2.1.8
- TM_RS00325 WP_004082568.1 19417 - 20461 (-) NC_000853.1:71049-72093 -
- TM_RS00330 WP_004082579.1 20680 - 22558 (+) NC_000853.1:72312-74190 -
- TM_RS00335 WP_004082581.1 22634 - 23642 (+) NC_000853.1:74266-75274 -
- TM_RS00340 WP_004082583.1 23658 - 24513 (+) NC_000853.1:75290-76145 -
- TM_RS00345 WP_041426669.1 24514 - 25522 (+) NC_000853.1:76146-77154 -
- TM_RS00350 WP_004082591.1 25508 - 26471 (+) NC_000853.1:77140-78103 -
- TM_RS00355 WP_004082594.1 26508 - 28845 (+) NC_000853.1:78140-80477 3.2.1.21
- TM_RS00360 WP_004082599.1 28849 - 29827 (+) NC_000853.1:80481-81459 -

Cluster number

1

Gene name

Gene position

Gene type

Found by CGCFinder?

- 1 - 2025 (-) CAZyme: GH67 Yes
- 2082 - 4055 (-) TC: gnl|TC-DB|Q9WXR2|3.A.1.5.29 Yes
- 4069 - 5085 (-) TC: gnl|TC-DB|Q9WXR3|3.A.1.5.29 Yes
- 5082 - 6077 (-) TC: gnl|TC-DB|Q9WXR4|3.A.1.5.29 Yes
- 6091 - 7179 (-) TC: gnl|TC-DB|Q9WXR5|3.A.1.5.29 Yes
- 7208 - 8194 (-) TC: gnl|TC-DB|Q9WXR6|3.A.1.5.29 Yes
- 8513 - 11692 (+) CAZyme: GH10|CBM22|CBM9 Yes
- 11748 - 12287 (+) other Yes
- 12334 - 12780 (-) other Yes
uxaC 12792 - 14147 (-) other Yes
- 14249 - 14989 (+) TF: DBD-Pfam|HTH_11,DBD-SUPERFAMILY|0040266 Yes
- 15058 - 15669 (+) other Yes
- 15666 - 16685 (+) STP: STP|PfkB Yes
uxuB 16698 - 18317 (+) other Yes
uxuA 18314 - 19396 (+) other Yes
- 19418 - 20461 (-) CAZyme: CBM22|GH10 Yes
- 20681 - 22558 (+) TC: gnl|TC-DB|Q9WXS6|3.A.1.5.13 Yes
- 22635 - 23642 (+) TC: gnl|TC-DB|Q9WXS7|3.A.1.5.13 Yes
- 23659 - 24513 (+) TC: gnl|TC-DB|Q9WXS8|3.A.1.5.13 Yes
- 24515 - 25522 (+) TC: gnl|TC-DB|Q9WXS9|3.A.1.5.13 Yes
- 25509 - 26471 (+) TC: gnl|TC-DB|Q9WXT0|3.A.1.5.13 Yes
- 26509 - 28845 (+) CAZyme: GH3 Yes
- 28850 - 29827 (+) CAZyme: CE7 Yes
- 29810 - 29827 (-) TC: gnl|TC-DB|Q9RK11|3.A.1.14.13 Yes

PUL ID

PUL0390

PubMed

21255309, Microb Biotechnol. 2010 Jan;3(1):84-92. doi: 10.1111/j.1751-7915.2009.00150.x. Epub 2009 Sep 18.

Title

Hyperthermostable acetyl xylan esterase.

Author

Drzewiecki K, Angelov A, Ballschmiter M, Tiefenbach KJ, Sterner R, Liebl W

Abstract

An esterase which is encoded within a Thermotoga maritima chromosomal gene cluster for xylan degradation and utilization was characterized after heterologous expression of the corresponding gene in Escherichia coli and purification of the enzyme. The enzyme, designated AxeA, shares amino acid sequence similarity and its broad substrate specificity with the acetyl xylan esterase from Bacillus pumilus, the cephalosporin C deacetylase from Bacillus subtilis, and other (putative) esterases, allowing its classification as a member of carbohydrate esterase family 7. The recombinant enzyme displayed activity with p-nitrophenyl-acetate as well as with various acetylated sugar substrates such as glucose penta-acetate, acetylated oat spelts xylan and DMSO (dimethyl sulfoxide)-extracted beechwood xylan, and with cephalosporin C. Thermotoga maritima AxeA represents the most thermostable acetyl xylan esterase known to date. In a 10 min assay at its optimum pH of 6.5 the enzyme's activity peaked at 90 degrees C. The inactivation half-life of AxeA at a protein concentration of 0.3 microg microl(-1) in the absence of substrate was about 13 h at 98 degrees C and about 67 h at 90 degrees C. Differential scanning calorimetry analysis of the thermal stability of AxeA corroborated its extreme heat resistance. A multi-phasic unfolding behaviour was found, with two apparent exothermic peaks at approximately 100-104 degrees C and 107.5 degrees C. In accordance with the crystal structure, gel filtration analysis at ambient temperature revealed that the enzyme has as a homohexameric oligomerization state, but a dimeric form was also found.