Three novel rhamnogalacturonan I- pectins degrading enzymes from Aspergillus aculeatinus: Biochemical characterization and application potential

Lemaire, Adrien; Garzón, Catalina; Perrin, Aurore; Habrylo, Olivier; Trezel, Pauline; Bassard, Solène; Lefebvre, Valérie; Wuytswinkel, Olivier Van; Antalick, Guillaume; Pau-Roblot, Corinne; Pelloux, Jérôme

doi:10.1016/j.carbpol.2020.116752

Cited by 22 publications

(7 citation statements)

References 52 publications

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“…The coding sequence of TBL38 (Q8VY22) was codon-optimized for Pichia pastoris and synthesized without signal peptide in frame with His-tag in pPCIZ-αB by ProteoGenix (Schiltigheim, France). rTBL38 was produced in P. pastoris following the previously described protocol (Lemaire et al, 2020). Activity assays of rTBL38 was performed with the acetic acid assay kit (K-ACETRM, Megazyme) using the activity buffer at 40°C and with three different acetylated substrates: 100 mM Triacetine, 10 mg.ml −1 xylan (24 % acetylation), and 10 mg.mL − 1 sugar beet pectins (31 % acetylation).…”

Section: Methodsmentioning

confidence: 99%

TBL38 is an atypical homogalacturonan acetylesterase with a peculiar cell wall microdomain localization in Arabidopsis seed mucilage secretory cells

Dauphin,

Ropartz,

Ranocha

et al. 2023

Preprint

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Plant cell walls are made of complex polysaccharidic/proteinaceous network whose biosynthesis and dynamics implicate several cell compartments and impact plant development. The synthesis and remodeling of homogalacturonan pectins is associated with multiple developmental processes ranging from growth to response to biotic/abiotic stress. It encompasses Golgi-localized methylation and acetylation and subsequent demethylation and deacetylation in the cell wall. In the last decade, our comprehension of plant polysaccharides acetylation has increased significantly thanks to the study of the TRICHOME BIREFRINGENCE-LIKE (TBL) protein family. TBLs are mostly described as Golgi-localized acetyltransferases specifically targeting diverse hemicelluloses or pectins. Various tbl mutants showed altered wall mechanical properties and dynamics. Here, we study TBL38 that is co-expressed with PECTIN METHYLESTERASE INHIBITOR6 (PMEI6) and PEROXIDASE 36 (PRX36) during the development of Arabidopsis seed mucilage secretory cells (MSCs). We demonstrate the atypical TBL38 cell wall localization restricted to the PMEI6/PRX36 MSC cell wall microdomain. A tbl38 mutant displays an intriguing homogalacturonan immunological phenotype in this cell wall microdomain and in a MSC surface-enriched abrasion powder. This fraction was further characterized by mass spectrometry oligosaccharide profiling revealing an increased homogalacturonan acetylation phenotype. Finally, a recombinant TBL38 is shown to display pectin acetylesterase activity in vitro. These results indicate that TBL38 is an atypical cell wall-localized TBL that displays a homogalacturonan acetylesterase activity rather than a Golgi-localized acetyltransferase activity as observed in previously studied TBLs. TBL38 function during seed development is discussed.

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Section: Methodsmentioning

confidence: 99%

TBL38 is an atypical homogalacturonan acetylesterase with a peculiar cell wall microdomain localization in Arabidopsis seed mucilage secretory cells

Dauphin,

Ropartz,

Ranocha

et al. 2023

Preprint

Self Cite

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“…Collected mycelium was frozen in liquid nitrogen, lyophilized and ground. Isolation of RNA and cDNA synthesis was realized as previously described in Lemaire et al [36].…”

Section: Fungal Strain and Growthmentioning

confidence: 99%

The structural, dynamical and biochemical characterizations ofVerticillium dahliaepectate lyase, VdPelB, highlight its specificities

Safran

Ung

Bouckaert

et al. 2022

Preprint

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Pectins, complex polysaccharides and major components of the plant primary cell wall, can be degraded by pectate lyases (PLs). PLs cleave glycosidic bonds of homogalacturonans (HG), the main pectic domain, by beta-elimination, releasing unsaturated oligogalacturonides (OGs). To understand the catalytic mechanism and structure/function of these enzymes, we characterized VdPelB from Verticillium dahliae, a plant pathogen. We first solved the crystal structure of VdPelB at 1.2A resolution showing that it is a right-handed parallel beta-helix structure. Molecular dynamics (MD) simulations further highlighted the dynamics of the enzyme in complex with substrates that vary in their degree of methylesterification, identifying amino acids involved in substrate binding and cleavage of non-methylesterified pectins. We then biochemically characterized wild type and mutated forms of VdPelB. VdPelB was most active on non-methylesterified pectins, at pH 8 in presence of Ca2+ ions. VdPelB-G125R mutant was most active at pH 9 and showed higher relative activity compared to native enzyme. The OGs released by VdPelB differed to that of previously characterized PLs, showing its peculiar specificity in relation to its structure. OGs released from Verticillium-partially tolerant and sensitive flax cultivars differed which could facilitate the identification VdPelB-mediated elicitors of defence responses.

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“…Methoxy esters, located in the C6 carboxyl groups of D-galacturonic acid, are substitutions generally found in the HG region and play an important role in the known functional properties and health benefits of these pectic polysaccharides [ 42 ]. RG-I is the second most abundant pectic polysaccharide in the cell wall of plants [ 43 ]. It is composed of a repeating backbone of galacturonic acid and rhamnose disaccharide, usually with neutral side chains [ 44 ].…”

Section: Polysaccharides From Agricultural Wastementioning

confidence: 99%

Strategies to Increase the Biological and Biotechnological Value of Polysaccharides from Agricultural Waste for Application in Healthy Nutrition

Rivas

Casquete

Martı́n

et al. 2021

IJERPH

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Nowadays, there is a growing interest in the extraction and identification of new high added-value compounds from the agro-food industry that will valorize the great amount of by-products generated. Many of these bioactive compounds have shown beneficial effects for humans in terms of disease prevention, but they are also of great interest in the food industry due to their effect of extending the shelf life of foods by their well-known antioxidant and antimicrobial activity. For this reason, an additional research objective is to establish the best conditions for obtaining these compounds from complex by-product structures without altering their activity or even increasing it. This review highlights recent work on the identification and characterization of bioactive compounds from vegetable by-products, their functional activity, new methodologies for the extraction of bioactive compounds from vegetables, possibly increasing their biological activity, and the future of the global functional food and nutraceuticals market.

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Three novel rhamnogalacturonan I- pectins degrading enzymes from Aspergillus aculeatinus: Biochemical characterization and application potential

Cited by 22 publications

References 52 publications

TBL38 is an atypical homogalacturonan acetylesterase with a peculiar cell wall microdomain localization in Arabidopsis seed mucilage secretory cells

TBL38 is an atypical homogalacturonan acetylesterase with a peculiar cell wall microdomain localization in Arabidopsis seed mucilage secretory cells

The structural, dynamical and biochemical characterizations ofVerticillium dahliaepectate lyase, VdPelB, highlight its specificities

Strategies to Increase the Biological and Biotechnological Value of Polysaccharides from Agricultural Waste for Application in Healthy Nutrition

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