Dataset Information

Identification and characterization of a hyperthermophilic GH9 cellulase from the Arctic Mid-Ocean Ridge vent field.

ABSTRACT: A novel GH9 cellulase (AMOR_GH9A) was discovered by sequence-based mining of a unique metagenomic dataset collected at the Jan Mayen hydrothermal vent field. AMOR_GH9A comprises a signal peptide, a catalytic domain and a CBM3 cellulose-binding module. AMOR_GH9A is an exceptionally stable enzyme with a temperature optimum around 100°C and an apparent melting temperature of 105°C. The novel cellulase retains 64% of its activity after 4 hours of incubation at 95°C. The closest characterized homolog of AMOR_GH9A is TfCel9A, a processive endocellulase from the model thermophilic bacterium Thermobifida fusca (64.2% sequence identity). Direct comparison of AMOR_GH9A and TfCel9A revealed that AMOR_GH9A possesses higher activity on soluble and amorphous substrates (phosphoric acid swollen cellulose, konjac glucomannan) and has an ability to hydrolyse xylan that is lacking in TfCel9A.

SUBMITTER: Stepnov AA

PROVIDER: S-EPMC6731012 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

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Identification and characterization of a hyperthermophilic GH9 cellulase from the Arctic Mid-Ocean Ridge vent field.

Stepnov Anton A AA Fredriksen Lasse L Steen Ida H IH Stokke Runar R Eijsink Vincent G H VGH

PloS one 20190906 9

A novel GH9 cellulase (AMOR_GH9A) was discovered by sequence-based mining of a unique metagenomic dataset collected at the Jan Mayen hydrothermal vent field. AMOR_GH9A comprises a signal peptide, a catalytic domain and a CBM3 cellulose-binding module. AMOR_GH9A is an exceptionally stable enzyme with a temperature optimum around 100°C and an apparent melting temperature of 105°C. The novel cellulase retains 64% of its activity after 4 hours of incubation at 95°C. The closest characterized homolog ...[more]

PMID: 31491027

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Project description:Textures and compositions of minerals can be used to infer the physiochemical conditions present within magmatic systems. Given that plagioclase is an abundant phase in many magmatic systems, understanding the link between texture and process is vital. Here, we present a database of textural and compositional data for?>?1800 plagioclase crystals in mid-ocean ridge basalt from the Gakkel Ridge (Arctic Ocean) to investigate the physiochemical conditions and processes that govern the formation of plagioclase textures and compositions. The Gakkel basalts have high modal crystal contents (up to 50%). The crystal cargo is complex, with both individual plagioclase and glomerocrysts showing large variations in crystal habit, zoning and resorption. The most common types of zoning are reverse and patchy; we attribute patchy zoning to infilling following either skeletal growth or resorption. Resorption is abundant, with multiple resorption events commonly present in a single crystal, and results from both magmatic recharge and decompression. Periods of strong undercooling, distinct to quench crystallisation, are indicated by matured skeletal crystals and thin normally zoned melt inclusion-rich bands following resorption. Individual samples often contain diverse textural and compositional plagioclase groups. Furthermore, most plagioclase is not in equilibrium with its host melt. Finally, the porous open structures of some glomerocrysts suggest that they represent pieces of entrained disaggregated mush. We interpret this to indicate that the crystal cargo is not generally phenocrystic in origin. Instead, plagioclase crystals that formed in different parts of a mush-dominated plumbing system were entrained into ascending melts. The textures of individual crystals are a function of their respective histories of (under)cooling, magma mixing and decompression. The morphologies of melt inclusion trapped in the plagioclase crystals are associated with specific host crystal textures, suggesting a link between plagioclase crystallisation processes and melt inclusion entrapment. The database of plagioclase presented herein may serve as a template for the interpretation of plagioclase textures in magmatic systems elsewhere.

Project description:A two-domain GH10 xylanase-encoding gene (amor_gh10a) was discovered from a metagenomic data set, generated after in situ incubation of a lignocellulosic substrate in hot sediments on the sea floor of the Arctic Mid-Ocean Ridge (AMOR). AMOR_GH10A comprises a signal peptide, a carbohydrate-binding module belonging to a previously uncharacterized family, and a catalytic glycosyl hydrolase (GH10) domain. The enzyme shares the highest sequence identity (42%) with a hypothetical protein from a Verrucomicrobia bacterium, and its GH10 domain shares low identity (24 to 28%) with functionally characterized xylanases. Purified AMOR_GH10A showed thermophilic and halophilic properties and was active toward various xylans. Uniquely, the enzyme showed high activity toward amorphous cellulose, glucomannan, and xyloglucan and was more active toward cellopentaose than toward xylopentaose. Binding assays showed that the N-terminal domain of this broad-specificity GH10 binds strongly to amorphous cellulose, as well as to microcrystalline cellulose, birchwood glucuronoxylan, barley β-glucan, and konjac glucomannan, confirming its classification as a novel CBM (CBM85).IMPORTANCE Hot springs at the sea bottom harbor unique biodiversity and are a promising source of enzymes with interesting properties. We describe the functional characterization of a thermophilic and halophilic multidomain xylanase originating from the Arctic Mid-Ocean Ridge vent system, belonging to the well-studied family 10 of glycosyl hydrolases (GH10). This xylanase, AMOR_GH10A, has a surprisingly wide substrate range and is more active toward cellopentaose than toward xylopentaose. This substrate promiscuity is unique for the GH10 family and could prove useful in industrial applications. Emphasizing the versatility of AMOR_GH10A, its N-terminal domain binds to both xylans and glycans, while not showing significant sequence similarities to any known carbohydrate-binding module (CBM) in the CAZy database. Thus, this N-terminal domain lays the foundation for the new CBM85 family.

Dataset Information

Identification and characterization of a hyperthermophilic GH9 cellulase from the Arctic Mid-Ocean Ridge vent field.

Publications

Identification and characterization of a hyperthermophilic GH9 cellulase from the Arctic Mid-Ocean Ridge vent field.

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