Fervidibacter sacchariis an aerobic hyperthermophile belonging to the phylumArmatimonadotathat degrades a variety of polysaccharides. Its genome encodes 117 enzymes with one or more annotated glycoside hydrolase (GH) domain, but the roles of these putative GHs in polysaccharide catabolism are poorly defined. Here, we describe oneF. saccharienzyme encoding a GH10 domain, Fsa02490Xyn, that was previously shown to be active onMiscanthus, oat β‐glucan, and beech‐wood xylan, with optimal activity at 90–100 °C. We show that Fsa02490Xyn is also active on birch‐wood xylan and gellan gum. The pH range on beech‐wood xylan was 4.5 to 9.5 (pHopt7.0–8.0). Fsa024940Xyn had aKmof 2.375 mm,Vmaxof 1250 μm·min−1, andkcat/Kmof 1.259 × 104 s−1·m−1when using apara‐nitrophenyl‐𝛽‐xylobioside assay. A phylogenetic analysis of GH10 family enzymes revealed a large clade of enzymes from diverse members of the classFervidibacteria, including Fsa02490Xyn and a second enzyme fromF. sacchari, with apparent horizontal gene transfer withinFervidibacteriaand betweenFervidibacteriaand thermophilicBacillota. This study establishes Fsa02490Xyn as a hyperthermophilic GH10 enzyme with endo‐β‐1,4‐xylanase activity and identifies a large clade of homologous GH10 enzymes within the classFervidibacteria. Impact statementThe depolymerization of xylan at high temperatures is important because this process limits the degradation of polysaccharides in nature and the synthesis of biofuels from plant wastes. Our study is also important becauseF. sacchariis one of only a few cultivated members of theArmatimonadota, which are polysaccharide‐degradation specialists.
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Hydrolysis of ionic liquid–treated substrate with an Iocasia fonsfrigidae strain SP3-1 endoglucanase
AbstractRecently, we reported the discovery of a novel endoglucanase of the glycoside hydrolase family 12 (GH12), designated IfCelS12A, from the haloalkaliphilic anaerobic bacteriumIocasia fonsfrigidaestrain SP3-1, which was isolated from a hypersaline pond in the Samut Sakhon province of Thailand (ca. 2017). IfCelS12A exhibits high substrate specificity on carboxymethyl cellulose and amorphous cellulose but low substrate specificity on b-1,3;1,4-glucan. Unlike some endoglucanases of the GH12 family, IfCelS12A does not exhibit hydrolytic activity on crystalline cellulose (i.e., Avicel™). High-Pressure Liquid Chromatography (HPLC) and Thin Layer Chromatography (TLC) analyses of products resulting from IfCelS12-mediated hydrolysis indicate mode of action for this enzyme. Notably, IfCelS12A preferentially hydrolyzes cellotetraoses, cellopentaoses, and cellohexaoses with negligible activity on cellobiose or cellotriose. Kinetic analysis with cellopentaose and barely b-d-glucan as cellulosic substrates were conducted. On cellopentaose, IfCelS12A demonstrates a 16-fold increase in activity (KM = 0.27 mM;kcat = 0.36 s−1;kcat/KM = 1.34 mM−1s−1) compared to the enzymatic hydrolysis of barley b-d-glucan (KM: 0.04 mM,kcat: 0.51 s−1,kcat/KM = 0.08 mM−1s−1). Moreover, IfCelS12A enzymatic efficacy is stable in hypersaline sodium chlorids (NaCl) solutions (up to 10% NaCl). Specifically, IfCel12A retains notable activity after 24 h at 2M NaCl (10% saline solution). IfCelS12A used as a cocktail component with other cellulolytic enzymes and in conjunction with mobile sequestration platform technology offers additional options for deconstruction of ionic liquid–pretreated cellulosic feedstock. Key points•IfCelS12A from an anaerobic alkaliphile Iocasia fronsfrigidae shows salt tolerance•IfCelS12A in cocktails with other enzymes efficiently degrades cellulosic biomass•IfCelS12A used with mobile enzyme sequestration platforms enhances hydrolysis
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- PAR ID:
- 10496105
- Editor(s):
- de Leon, Lester
- Publisher / Repository:
- Spring Nature
- Date Published:
- Journal Name:
- Applied Microbiology and Biotechnology
- Edition / Version:
- 1
- Volume:
- 108
- Issue:
- 1
- ISSN:
- 0175-7598
- Subject(s) / Keyword(s):
- Iocasia fronsfrigidae IfCelS12A halophile MESP
- Format(s):
- Medium: X Size: 1.7MB Other: pdf
- Size(s):
- 1.7MB
- Sponsoring Org:
- National Science Foundation
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