Identification of novel enzymes to enhance the ruminal digestion of barley straw

Badhan, Ajay; Ribeiro, Gabriel de Oliveira; Jones, Darryl R.; Wang, Yuxi; Abbott, D. Wade; Falco, Marcos Di; Tsang, Adrian; McAllister, Tim A.

doi:10.1016/j.biortech.2018.03.086

Cited by 16 publications

(12 citation statements)

References 42 publications

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“…Proteins were separated by SDS-PAGE until the 250-kDa marker of the BLUelf prestained protein ladder (FroggaBio, North York, Toronto, Canada) entered the separating gel. After the gel was stained with Coomassie brilliant blue, about 1 cm of the gel lane was cut out and polypeptides were in-gel digested with trypsin as described previously [63]. Peptide mixtures were resuspended in 45 μL 5% (v/v) aqueous acetonitrile with 0.1% (v/v) formic acid, containing 4 fmol/μL predigested bovine serum albumin.…”

Section: Secretome Extraction Protein Preparation and Analysismentioning

confidence: 99%

Evidence for ligninolytic activity of the ascomycete fungus Podospora anserina

Erven

Kleijn

Patyshakuliyeva

et al. 2020

Biotechnol Biofuels

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Background: The ascomycete fungus Podospora anserina has been appreciated for its targeted carbohydrate-active enzymatic arsenal. As a late colonizer of herbivorous dung, the fungus acts specifically on the more recalcitrant fraction of lignocellulose and this lignin-rich biotope might have resulted in the evolution of ligninolytic activities. However, the lignin-degrading abilities of the fungus have not been demonstrated by chemical analyses at the molecular level and are, thus far, solely based on genome and secretome predictions. To evaluate whether P. anserina might provide a novel source of lignin-active enzymes to tap into for potential biotechnological applications, we comprehensively mapped wheat straw lignin during fungal growth and characterized the fungal secretome.Results: Quantitative 13 C lignin internal standard py-GC-MS analysis showed substantial lignin removal during the 7 days of fungal growth (24% w/w), though carbohydrates were preferably targeted (58% w/w removal). Structural characterization of residual lignin by using py-GC-MS and HSQC NMR analyses demonstrated that C α -oxidized substructures significantly increased through fungal action, while intact β-O-4′ aryl ether linkages, p-coumarate and ferulate moieties decreased, albeit to lesser extents than observed for the action of basidiomycetes. Proteomic analysis indicated that the presence of lignin induced considerable changes in the secretome of P. anserina. This was particularly reflected in a strong reduction of cellulases and galactomannanases, while H 2 O 2 -producing enzymes clearly increased. The latter enzymes, together with laccases, were likely involved in the observed ligninolysis. Conclusions:For the first time, we provide unambiguous evidence for the ligninolytic activity of the ascomycete fungus P. anserina and expand the view on its enzymatic repertoire beyond carbohydrate degradation. Our results can be of significance for the development of biological lignin conversion technologies by contributing to the quest for novel lignin-active enzymes and organisms.

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Section: Secretome Extraction Protein Preparation and Analysismentioning

confidence: 99%

Evidence for ligninolytic activity of the ascomycete fungus Podospora anserina

Erven

Kleijn

Patyshakuliyeva

et al. 2020

Biotechnol Biofuels

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“…The use of the fibrous fraction of feed by ruminants is related to the ability of rumen microorganisms to synthesize and secrete enzymes that hydrolyze the cell walls of plants. However, in diets having a high fiber and dry matter content, the use of feed is inefficient [1]. One of the solutions to try to solve part of this problem is the use of exogenous fibrolytic enzymes in the feed of ruminants, especially in ensiled materials.…”

Section: Introductionmentioning

confidence: 99%

Reduction in lignin content and increase in the antioxidant capacity of corn and sugarcane silages treated with an enzymatic complex produced by white rot fungus

et al. 2020

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The objective was to evaluate the effect of the addition of 0, 10, 20, and 30 mg.kg-1 of natural matter of a lignocellulosic enzymatic complex produced by the white rot fungus on the chemical composition, cumulative gas production in vitro, and antioxidant compounds of corn and sugarcane silages. After being chopped and treated with the enzymatic complex, the plants were packed in vacuum-sealed bags. After 60 days, the mini silos were opened and the samples were dried in a forced ventilation oven at 55˚C for analysis of the proposed parameters. The experiment was conducted in a completely randomized design with four replicates per treatment. In the corn silage, there was a linear reduction in the lignin concentration. In the sugarcane silage showed a reduction of 12% in the lignin concentration, a linear reduction in the hemicellulose content, and a decrease of 8% in the cellulose concentration compared to the control treatment. The lignin monomers had linear increases in the syringyl:guaiacil ratio. This reflected on significant increases in the concentration of the non-fibrous carbohydrates and the A + B1 fraction of the carbohydrates, and a reduction in the C fraction. The in vitro gas production increased, the time of colonization and initiation of in vitro fermentation linearly decreased in both silages. The phenolic compounds and the antioxidant capacity increased linearly with the addition of the enzymes in both silages. The addition of the lignocellulolytic enzymes to the silages caused changes in the cell wall, resulting in improvements in the in vitro fermentative parameters, besides the additional effect on the antioxidant capacity. There was an effect of the addition of the enzymes on the evaluated fodder, and the best concentration was, on average, 20 mg kg-1 MN for corn silage and 10 mg kg-1 NM for sugarcane silage.

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“…The most efficient enzymatic extraction was performed with Viscozyme L. The Viscozyme L preparation includes a diversity of enzymes and the higher pectin yields compared to Pectinase treatments suggest that multiple activities are required for an efficient pectin extraction from SBP (Figure A) …”

Section: Methodsmentioning

confidence: 99%

“…In pectin extracted with Viscozyme L, the hydrolysis starts during the extraction due to the side activity of this enzymatic cocktail. The esterase activity of the Viscozyme L cocktail may therefore make the pectin more susceptible to glycosyl hydrolases and lyases after de‐esterification . Although the chemical hydrolysis of microwave PEM showed the highest GalA yield, enzymatic hydrolysis produces large amounts of Ara that can be a valuable side‐product while yielding >80% of GalA.…”

Section: Methodsmentioning

confidence: 99%

Sustainable Galactarate‐Based Polymers: Multi‐Enzymatic Production of Pectin‐Derived Polyesters

Vastano

Pellis

Machado

et al. 2019

Macromol. Rapid Commun.

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Large amounts of agricultural wastes are rich in pectins that, in many cases, disrupt the processing of food residues due to gelation. Despite pectins being a promising sustainable feedstock for bio‐based chemical production, the current pathways to produce platform molecules from this polysaccharide are hazardous and entail the use of strong acids. The present work describes a sequence of biocatalyzed reactions that involves 1) the extraction of pectin from sugar beet pulp and enzymatic recovery of galacturonic acid (GalA), followed by 2) the enzymatic oxidation of the GalA aldehyde and the recovery of galactaric acid (GA), and 3) the biocatalyzed polycondensation of GA to obtain fully bio‐based polyesters carrying lateral hydroxy functionalities. The acid‐free pectin extraction is optimized using enzymes and microwave technology. The conditions for enzymatic oxidation of GalA allow the separation of the GA produced by a simple centrifugation step that leads to the enzyme‐catalyzed polycondensation reactions.

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Identification of novel enzymes to enhance the ruminal digestion of barley straw

Cited by 16 publications

References 42 publications

Evidence for ligninolytic activity of the ascomycete fungus Podospora anserina

Evidence for ligninolytic activity of the ascomycete fungus Podospora anserina

Reduction in lignin content and increase in the antioxidant capacity of corn and sugarcane silages treated with an enzymatic complex produced by white rot fungus

Sustainable Galactarate‐Based Polymers: Multi‐Enzymatic Production of Pectin‐Derived Polyesters

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