Dataset Information

Activity-based protein profiling of xylan-degrading glycoside hydrolases in Aspergillus secretomes

ABSTRACT: Plant polysaccharides represent a virtually unlimited feedstock for the generation of biofuels and other commodities. However, the extraordinary recalcitrance of plant polysaccharides towards breakdown necessitates a continued search for enzymes that degrade these materials efficiently under defined conditions. Activity-based protein profiling (ABPP) provides a route for the functional discovery of such enzymes in complex mixtures and under industrially relevant conditions. Here, we show the detection and identification of b-xylosidases and endo-xylanases in the secretome of Aspergillus niger, by the use of chemical probes inspired by the b-glucosidase inhibitor cyclophellitol. Furthermore, we demonstrate the use of these activity-based probes (ABPs) to assess enzyme-substrate specificities, thermal stabilities, and other biotechnologically relevant parameters. Our experiments highlight the utility of ABPs as promising tools for the discovery of relevant enzymes useful for biomass breakdown.

INSTRUMENT(S): maXis

ORGANISM(S): Aspergillus Niger (ncbitaxon:5061)

SUBMITTER: Gideon J. Davies

PROVIDER: MSV000083714 | MassIVE | Wed Apr 24 04:09:00 BST 2019

SECONDARY ACCESSION(S): PXD013618

REPOSITORIES: MassIVE

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Publications

Dynamic and Functional Profiling of Xylan-Degrading Enzymes in <i>Aspergillus</i> Secretomes Using Activity-Based Probes.

Schröder Sybrin P SP de Boer Casper C McGregor Nicholas G S NGS Rowland Rhianna J RJ Moroz Olga O Blagova Elena E Reijngoud Jos J Arentshorst Mark M Osborn David D Morant Marc D MD Abbate Eric E Stringer Mary A MA Krogh Kristian B R M KBRM Raich Lluís L Rovira Carme C Berrin Jean-Guy JG van Wezel Gilles P GP Ram Arthur F J AFJ Florea Bogdan I BI van der Marel Gijsbert A GA Codée Jeroen D C JDC Wilson Keith S KS Wu Liang L Davies Gideon J GJ Overkleeft Herman S HS

ACS central science 20190524 6

Plant polysaccharides represent a virtually unlimited feedstock for the generation of biofuels and other commodities. However, the extraordinary recalcitrance of plant polysaccharides toward breakdown necessitates a continued search for enzymes that degrade these materials efficiently under defined conditions. Activity-based protein profiling provides a route for the functional discovery of such enzymes in complex mixtures and under industrially relevant conditions. Here, we show the detection a ...[more]

PMID: 31263766

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Project description:Background: Dendrobium officinale, an endangered Chinese herb, has extensive therapeutic effects and contains bioactive ingredients including a large number of polysaccharides and alkaloids, and minimal flavonoids. Firstly, this study attempts to obtain the protocorm-like bodies of this plant through tissue culture to produce the main secondary metabolites whose distribution in each organelle and protocorm like bodies is analyzed. Then, analysis of the correlation between comparative transcriptome sequence and the metabolite content in different organs enables the discovery of putative genes encoding enzymes involved in the biosynthesis of polysaccharides and alkaloids, and flavonoids. Results: The optimum condition for protocorm-like bodies (PLBs) induction and propagation of D. officinale is established. For protocorm induction, we use the seed as the explant, and the optimum medium formula for PLBs propagation is 1/2 MS + α-NAA 0.5 mg·L-1 +6-BA 1.0 mg·L-1 + 2, 4-D 1.5-2.0 mg·L-1 + potato juice 100 g·L-1. The distribution of polysaccharides, alkaloids and flavonoids in D. officinale organs was clarified. Stems, PLBs and leaves have the highest content of polysaccharides, alkaloids and flavonoids, respectively. PLBs replace organs to produce alkaloids in D. officinale, and naringenin was only produced in stem. Hot water extraction (HWE) method was found outperforming the ultrasound-assisted extraction (UAE) method for polysaccharides from D. officinale. A comparative transcriptome analysis of the protocorm-like bodies and leaves of D. officinale showed genes encoding enzymes involved in polysaccharides, alkaloids and flavonoids biosynthetic pathway were differentially expressed. Putative genes encoding enzymes involved in polysaccharides, alkaloids and flavonoids synthetic pathway were identified. Notably, genes encoding enzymes of strictosidine beta-glucosidase, geissoschizine synthase and vinorine synthase in alkaloids biosynthesis of D. officinale are first reported. Conclusions: Our works, especially the identification of candidate genes encoding enzymes involved in metabolites biosynthesis will help to explore and protect the endangered genetic resources and will also facilitate further analysis of the molecular mechanism of secondary metabolites’ biosynthesis in D. officinale.

Dataset Information

Activity-based protein profiling of xylan-degrading glycoside hydrolases in Aspergillus secretomes

Publications

Dynamic and Functional Profiling of Xylan-Degrading Enzymes in <i>Aspergillus</i> Secretomes Using Activity-Based Probes.

Similar Datasets

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