A novel member of glycoside hydrolase family 30 subfamily 8 with altered substrate specificity

John, Franz J. St; Dietrich, Diane; Crooks, Casey; Pozharski, Edwin; González, Javier Martı́nez; Bales, Elizabeth; Smith, Kennon M.; Hurlbert, Jason C.

doi:10.1107/s1399004714019531

Cited by 28 publications

(28 citation statements)

References 36 publications

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“…This is combined with lower selectivity for unsubstituted xylan, AX, and XOS, which may be structurally reflected by more open-binding clefts (Sainz-Polo et al 2014 ). (At least one exception from this rule is however known as the enzyme from C. papyrosolvens (St John et al 2014 ) is active on non-substituted xylan).…”

Section: Gh30 Glucuronoarabinoxylan Endoxylanasesmentioning

confidence: 99%

Endo-xylanases as tools for production of substituted xylooligosaccharides with prebiotic properties

Karlsson

Schmitz

Linares-Pastén

et al. 2018

Appl Microbiol Biotechnol

120

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Xylan has a main chain consisting of β-1,4-linked xylose residues with diverse substituents. Endoxylanases cleave the xylan chain at cleavage sites determined by the substitution pattern and thus give different oligosaccharide product patterns. Most known endoxylanases belong to glycoside hydrolase (GH) families 10 and 11. These enzymes work well on unsubstituted xylan but accept substituents in certain subsites. The GH11 enzymes are more restricted by substituents, but on the other hand, they are normally more active than the GH10 enzymes on insoluble substrates, because of their smaller size. GH5 endoxylanases accept arabinose substituents in several subsites and require it in the − 1 subsite. This specificity makes the GH5 endoxylanases very useful for degradation of highly arabinose-substituted xylans and for the selective production of arabinoxylooligosaccharides, without formation of unsubstituted xylooligosaccharides. The GH30 endoxylanases have a related type of specificity in that they require a uronic acid substituent in the − 2 subsite, which makes them very useful for the production of uronic acid substituted oligosaccharides. The ability of dietary xylooligosaccharides to function as prebiotics in humans is governed by their substitution patterns. Endoxylanases are thus excellent tools to tailor prebiotic oligosaccharides to stimulate various types of intestinal bacteria and to cause fermentation in different parts of the gastrointestinal tract. Continuously increasing knowledge on the function of the gut microbiota and discoveries of novel endoxylanases increase the possibilities to achieve health-promoting effects.

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Section: Gh30 Glucuronoarabinoxylan Endoxylanasesmentioning

confidence: 99%

Endo-xylanases as tools for production of substituted xylooligosaccharides with prebiotic properties

Karlsson

Schmitz

Linares-Pastén

et al. 2018

Appl Microbiol Biotechnol

120

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“…Xylanase activity is mainly found in subfamily 8, the only exception being two characterized but not structure determined xylanases in subfamily 7 ( www.cazy.org ) [ 101 , 102 ]. Today (March 2016), four characterized GH30 xylanases are structure determined: BsXynC from Bacillus subtilis [ 103 , 104 ], EcXynA from Erwinia chrysanthemi [ 76 , 105 ], CpXyn30A from Clostridium papyrosolvens [ 106 ] and Xyn30D from Paenibacillus barcinonensis [ 107 ] as well as the not characterized Xyn30A from Clostridium thermocellum (PDB structures 4CKQ, 4UQ9 and 4UQA to AUQE).…”

Section: Gh30 Subfamily 8 Endo-xylanasesmentioning

confidence: 99%

Structural Considerations on the Use of Endo-Xylanases for the Production of prebiotic Xylooligosaccharides from Biomass

Linares-Pastén

Aronsson

Karlsson

2017

CPPS

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Xylooligosaccharides (XOS) have gained increased interest as prebiotics during the last years. XOS and arabinoxylooligosaccharides (AXOS) can be produced from major fractions of biomass including agricultural by-products and other low cost raw materials. Endo-xylanases are key enzymes for the production of (A)XOS from xylan. As the xylan structure is broadly diverse due to different substi-tutions, diverse endo-xylanases have evolved for its degradation. In this review structural and functional aspects are discussed, focusing on the potential applications of endo-xylanases in the production of dif-ferently substituted (A)XOS as emerging prebiotics, as well as their implication in the processing of the raw materials. Endo-xylanases are found in at least eight different glycoside hydrolase families (GH), and can either have a retaining or an inverting catalytic mechanism. To date, it is mainly retaining endo-xylanases that are used in applications to produce (A)XOS. Enzymes from these GH-families (mainly GH10 and GH11, and the more recently investigated GH30) are taken as prototypes to discuss substrate preferences and main products obtained. Finally, the need of new and accessory enzymes (new specifici-ties from new families or sources) to increase the yield of different types of (A)XOS is discussed, along with in vitro tests of produced oligosaccharides and production of enzymes in GRAS organisms to fa-cilitate use in functional food manufacturing.

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“…In addition, Xyn30D from Paenibacillus barcinonensis and Xyn30A from B. licheniformis SVD1 have been shown to display a high activity on glucuronoxylans, but no activity on linear xylooligomers (Valenzuela et al, 2012;Sakka et al, 2012). However, the recently solved structure of xylanase 30A from Clostridium papyrosolvens (CpC71) displays a low specificity for the GlcA appendage owing to a lack of the 8-8 loop region (St John et al, 2014). In addition, Xyn5B from Bacillus sp.…”

Section: Introductionmentioning

confidence: 99%

“…Here, we determined the structure of CtXyn30A in its native form and that of the CtXyn30A-E225A mutant bound to xylotetraose. Comparison of the GH30 structure with those of XynA from E. chrysanthemi (Urbá niková et al, 2011), XynC from B. subtilis (St John et al, 2011), CpC71 from C. papyrosolvens (St John et al, 2014) and XynD from P. barcinonensis (Sainz-Polo et al, 2014) revealed remarkable conservation in the catalytic machinery of both mesophilic and thermophilic glucuronoxylan endo--1,4-xylanases.…”

Section: Introductionmentioning

confidence: 99%

Conservation in the mechanism of glucuronoxylan hydrolysis revealed by the structure of glucuronoxylan xylanohydrolase (CtXyn30A) fromClostridium thermocellum

Freire

Verma

Bule

et al. 2016

Acta Cryst Sect D Struct Biol

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Glucuronoxylan endo-β-1,4-xylanases cleave the xylan chain specifically at sites containing 4-O-methylglucuronic acid substitutions. These enzymes have recently received considerable attention owing to their importance in the cooperative hydrolysis of heteropolysaccharides. However, little is known about the hydrolysis of glucuronoxylans in extreme environments. Here, the structure of a thermostable family 30 glucuronoxylan endo-β-1,4-xylanase (CtXyn30A) from Clostridium thermocellum is reported. CtXyn30A is part of the cellulosome, a highly elaborate multi-enzyme complex secreted by the bacterium to efficiently deconstruct plant cell-wall carbohydrates. CtXyn30A preferably hydrolyses glucuronoxylans and displays maximum activity at pH 6.0 and 70°C. The structure of CtXyn30A displays a (β/α) TIM-barrel core with a side-associated β-sheet domain. Structural analysis of the CtXyn30A mutant E225A, solved in the presence of xylotetraose, revealed xylotetraose-cleavage oligosaccharides partially occupying subsites -3 to +2. The sugar ring at the +1 subsite is held in place by hydrophobic stacking interactions between Tyr139 and Tyr200 and hydrogen bonds to the OH group of Tyr227. Although family 30 glycoside hydrolases are retaining enzymes, the xylopyranosyl ring at the -1 subsite of CtXyn30A-E225A appears in the α-anomeric configuration. A set of residues were found to be strictly conserved in glucuronoxylan endo-β-1,4-xylanases and constitute the molecular determinants of the restricted specificity displayed by these enzymes. CtXyn30A is the first thermostable glucuronoxylan endo-β-1,4-xylanase described to date. This work reveals that substrate recognition by both thermophilic and mesophilic glucuronoxylan endo-β-1,4-xylanases is modulated by a conserved set of residues.

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A novel member of glycoside hydrolase family 30 subfamily 8 with altered substrate specificity

Cited by 28 publications

References 36 publications

Endo-xylanases as tools for production of substituted xylooligosaccharides with prebiotic properties

Endo-xylanases as tools for production of substituted xylooligosaccharides with prebiotic properties

Structural Considerations on the Use of Endo-Xylanases for the Production of prebiotic Xylooligosaccharides from Biomass

Conservation in the mechanism of glucuronoxylan hydrolysis revealed by the structure of glucuronoxylan xylanohydrolase (CtXyn30A) fromClostridium thermocellum

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