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Combinatorial Glycomic Analyses to Direct CAZyme Discovery for the Tailored Degradation of Canola Meal Non-Starch Dietary Polysaccharides

Lookup NU author(s): Dr Catherine Tétard-Jones, Professor William WillatsORCiD

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This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).

Abstract

Canola meal (CM), the protein-rich by-product of canola oil extraction, has shown promise as an alternative feedstuff and protein supplement in poultry diets, yet its use has been limited due to the abundance of plant cell wall fibre, specifically non-starch polysaccharides (NSP) and lignin. The addition of exogenous enzymes to promote the digestion of CM NSP in chickens has potential to increase the metabolizable energy of CM. We isolated chicken cecal bacteria from a continuous-flow mini-bioreactor system and selected for those with the ability to metabolize CM NSP. Of 100 isolates identified, Bacteroides spp. and Enterococcus spp. were the most common species with these capabilities. To identify enzymes specifically for the digestion of CM NSP, we used a combination of glycomics techniques, including enzyme-linked immunosorbent assay characterization of the plant cell wall fractions, glycosidic linkage analysis (methylation-GC-MS analysis) of CM NSP and their fractions, bacterial growth profiles using minimal media supplemented with CM NSP, and the sequencing and de novo annotation of bacterial genomes of high-efficiency CM NSP utilizing bacteria. The SACCHARIS pipeline was used to select plant cell wall active enzymes for recombinant production and characterization. This approach represents a multidisciplinary innovation platform to bioprospect endogenous CAZymes from the intestinal microbiota of herbivorous and omnivorous animals which is adaptable to a variety of applications and dietary polysaccharides.

Publication metadata

Author(s): Low KE, Xing X, Moote PE, Inglis G, Venketachalam S, Hahn MG, Marissa LK, Tétard-Jones CY, Jones DR, Willats WGTW, Slominski BA, Abbott DW

Publication type: Article

Publication status: Published

Journal: Microorganisms

Year: 2020

Volume: 8

Issue: 12

Online publication date: 29/11/2020

Acceptance date: 24/11/2020

Date deposited: 17/12/2020

ISSN (electronic): 2076-2607

Publisher: MDPI AG

URL: https://doi.org/10.3390/microorganisms8121888

DOI: 10.3390/microorganisms8121888

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