Characterization of a Novel β-Xylosidase, XylC, from
            <i>Thermoanaerobacterium saccharolyticum</i>
            JW/SL-YS485

Shao, Weilan; Xue, Yemin; Wu, Ailian; Kataeva, Irina; Pei, Jianjun; Wu, Huawei; Wiegel, Juergen

doi:10.1128/aem.01511-10

Cited by 55 publications

(57 citation statements)

References 36 publications

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“…The only single gene deletion strain with a distinct phenotype during growth on xylan was the ␤-xylosidase xylD deletion mutant (⌬xylD), while the deletion of xylB had little effect on growth on xylan. Since XylB was not found as an active protein from cell extracts in previous studies (15,24), XylD is likely the major ␤-xylosidase utilized by T. saccharolyticum during growth on birchwood xylan. This observation is also supported by the accumulation of short-chain xylo-oligomers in the absence of xylD, suggesting that in this strain, the cleavage of xylobiose to xylose is the rate-limiting step for growth on xylan.…”

Section: Discussionmentioning

confidence: 99%

“…While both xylB and xylC have been identified in the genome, the third ␤-xylosidase, XylA, has been purified to apparent homogeneity only from T. saccharolyticum. The gene for this ␤-xylosidase has not yet been identified (24). In addition to the primary xylanase and ␤-xylosidase enzymes, several accessory enzymes, including two xylan acetyl esterases (23) and an ␣-glucuronidase (15,22), have been characterized.…”

mentioning

confidence: 99%

“…Two of these ␤-xylosidases, XylA and XylC, have been puri-fied as active enzymes from crude cell extract, and the sequence of XylC has been determined (24). The third ␤-xylosidase, XylB, has been characterized only as a recombinant enzyme (15).…”

mentioning

confidence: 99%

“…The third ␤-xylosidase, XylB, has been characterized only as a recombinant enzyme (15). The gene product of xylB results in a protein smaller than both XylA and XylC with unique kinetic properties (24). XylB is a family 39 glycoside hydrolase enzyme and an ortholog of the xynB gene from T. saccharolyticum B6A-RI (13).…”

mentioning

confidence: 99%

“…XylB is a family 39 glycoside hydrolase enzyme and an ortholog of the xynB gene from T. saccharolyticum B6A-RI (13). The recently identified XylC represents a novel glycoside hydrolase family, displaying unusual kinetics and an amino acid sequence lacking similarity to any of the previously described glycoside hydrolase families with xylosidase activities (families 3, 39, 43, 52, and 54; www.cazy.org/Glycoside -Hydrolases.html) (24). While both xylB and xylC have been identified in the genome, the third ␤-xylosidase, XylA, has been purified to apparent homogeneity only from T. saccharolyticum.…”

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confidence: 99%

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Characterization of Xylan Utilization and Discovery of a New Endoxylanase in Thermoanaerobacterium saccharolyticum through Targeted Gene Deletions

Podkaminer

Guss

Trajano

et al. 2012

Appl Environ Microbiol

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eThe economical production of fuels and commodity chemicals from lignocellulose requires the utilization of both the cellulose and hemicellulose fractions. Xylanase enzymes allow greater utilization of hemicellulose while also increasing cellulose hydrolysis. Recent metabolic engineering efforts have resulted in a strain of Thermoanaerobacterium saccharolyticum that can convert C 5 and C 6 sugars, as well as insoluble xylan, into ethanol at high yield. To better understand the process of xylan solubilization in this organism, a series of targeted deletions were constructed in the homoethanologenic T. saccharolyticum strain M0355 to characterize xylan hydrolysis and xylose utilization in this organism. While the deletion of ␤-xylosidase xylD slowed the growth of T. saccharolyticum on birchwood xylan and led to an accumulation of short-chain xylo-oligomers, no other single deletion, including the deletion of the previously characterized endoxylanase XynA, had a phenotype distinct from that of the wild type. This result indicates a multiplicity of xylanase enzymes which facilitate xylan degradation in T. saccharolyticum. Growth on xylan was prevented only when a previously uncharacterized endoxylanase encoded by xynC was also deleted in conjunction with xynA. Sequence analysis of xynC indicates that this enzyme, a low-molecular-weight endoxylanase with homology to glycoside hydrolase family 11 enzymes, is secreted yet untethered to the cell wall. Together, these observations expand our understanding of the enzymatic basis of xylan hydrolysis by T. saccharolyticum.

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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confidence: 99%

mentioning

confidence: 99%

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Characterization of Xylan Utilization and Discovery of a New Endoxylanase in Thermoanaerobacterium saccharolyticum through Targeted Gene Deletions

Podkaminer

Guss

Trajano

et al. 2012

Appl Environ Microbiol

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Esterase EstK from Pseudomonas putida mt‐2: An enantioselective acetylesterase with activity for deacetylation of xylan and poly(vinylacetate)

Millar

Rahmanpour

Yuan

et al. 2017

Biotech and App Biochem

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An extracellular esterase gene estK was identified in Pseudomonas putida mt‐2 and overexpressed at high levels in Escherichia coli. The recombinant EstK enzyme was purified and characterized kinetically against p‐nitrophenyl ester and other aryl–alkyl ester substrates and found to be selective for hydrolysis of acetyl ester substrates with high activity for p‐nitrophenyl acetate (kcat 5.5 Sec−1, KM 285 µM). Recombinant EstK was found to catalyze deacetylation of acetylated beech xylan, indicating a possible in vivo function for this enzyme, and partial deacetylation of a synthetic polymer (poly(vinylacetate)). EstK was found to catalyze enantioselective hydrolysis of racemic 1‐phenylethyl acetate, generating 1R‐phenylethanol with an enantiomeric excess of 80.4%.

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Structural and functional analyses of catalytic domain of GH10 xylanase from Thermoanaerobacterium saccharolyticum JW/SL‐YS485

et al. 2013

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Xylanases are capable of decomposing xylans, the major components in plant cell wall, and releasing the constituent sugars for further applications. Because xylanase is widely used in various manufacturing processes, high specific activity, and thermostability are desirable. Here, the wild-type and mutant (E146A and E251A) catalytic domain of xylanase from Thermoanaerobacterium saccharolyticum JW/SL-YS485 (TsXylA) were expressed in Escherichia coli and purified subsequently. The recombinant protein showed optimal temperature and pH of 75°C and 6.5, respectively, and it remained fully active even after heat treatment at 75°C for 1 h. Furthermore, the crystal structures of apo-form wild-type TsXylA and the xylobiose-, xylotriose-, and xylotetraose-bound E146A and E251A mutants were solved by X-ray diffraction to high resolution (1.32-1.66 Å). The protein forms a classic (β/α)8 folding of typical GH10 xylanases. The ligands in substrate-binding groove as well as the interactions between sugars and active-site residues were clearly elucidated by analyzing the complex structures. According to the structural analyses, TsXylA utilizes a double displacement catalytic machinery to carry out the enzymatic reactions. In conclusion, TsXylA is effective under industrially favored conditions, and our findings provide fundamental knowledge which may contribute to further enhancement of the enzyme performance through molecular engineering.

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Characterization of a Novel β-Xylosidase, XylC, from Thermoanaerobacterium saccharolyticum JW/SL-YS485

Cited by 55 publications

References 36 publications

Characterization of Xylan Utilization and Discovery of a New Endoxylanase in Thermoanaerobacterium saccharolyticum through Targeted Gene Deletions

Characterization of Xylan Utilization and Discovery of a New Endoxylanase in Thermoanaerobacterium saccharolyticum through Targeted Gene Deletions

Esterase EstK from Pseudomonas putida mt‐2: An enantioselective acetylesterase with activity for deacetylation of xylan and poly(vinylacetate)

Structural and functional analyses of catalytic domain of GH10 xylanase from Thermoanaerobacterium saccharolyticum JW/SL‐YS485

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