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

2020 ◽  
Vol 8 (12) ◽  
pp. 1888 ◽  
Author(s):  
Kristin E. Low ◽  
Xiaohui Xing ◽  
Paul E. Moote ◽  
G. Douglas Inglis ◽  
Sivasankari Venketachalam ◽  
...  
Keyword(s):  
Cell Wall ◽  
Plant Cell ◽  
Plant Cell Wall ◽  
High Efficiency ◽  
De Novo ◽  
Common Species ◽  
Metabolizable Energy ◽  
Canola Meal ◽  
Enzyme Linked Immunosorbent Assay ◽  
Poultry Diets

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.

scholarly journals The GH10 and GH48 dual-functional catalytic domains from a multimodular glycoside hydrolase synergize in hydrolyzing both cellulose and xylan

2019 ◽  
Vol 12 (1) ◽  
Author(s):  
Yindi Chu ◽  
Zhenzhen Hao ◽  
Kaikai Wang ◽  
Tao Tu ◽  
Huoqing Huang ◽  
...  
Keyword(s):  
Cell Wall ◽  
Plant Cell ◽  
Plant Cell Wall ◽  
High Efficiency ◽  
Early Stage ◽  
Glycoside Hydrolases ◽  
Cell Wall Polysaccharides ◽  
Catalytic Domains ◽  
Xylan Hydrolysis ◽  
Dual Functional

Abstract Background Regarding plant cell wall polysaccharides degradation, multimodular glycoside hydrolases (GHs) with two catalytic domains separated by one or multiple carbohydrate-binding domains are rare in nature. This special mode of domain organization endows the Caldicellulosiruptor bescii CelA (GH9-CBM3c-CBM3b-CBM3b-GH48) remarkably high efficiency in hydrolyzing cellulose. CbXyn10C/Cel48B from the same bacterium is also such an enzyme which has, however, evolved to target both xylan and cellulose. Intriguingly, the GH10 endoxylanase and GH48 cellobiohydrolase domains are both dual functional, raising the question if they can act synergistically in hydrolyzing cellulose and xylan, the two major components of plant cell wall. Results In this study, we discovered that CbXyn10C and CbCel48B, which stood for the N- and C-terminal catalytic domains, respectively, cooperatively released much more cellobiose and cellotriose from cellulose. In addition, they displayed intramolecular synergy but only at the early stage of xylan hydrolysis by generating higher amounts of xylooligosaccharides including xylotriose, xylotetraose, and xylobiose. When complex lignocellulose corn straw was used as the substrate, the synergy was found only for cellulose but not xylan hydrolysis. Conclusion This is the first report to reveal the synergy between a GH10 and a GH48 domain. The synergy discovered in this study is helpful for understanding how C. bescii captures energy from these recalcitrant plant cell wall polysaccharides. The insight also sheds light on designing robust and multi-functional enzymes for plant cell wall polysaccharides degradation.

scholarly journals The Endo-Arabinanase BcAra1 Is a Novel Host-Specific Virulence Factor of the Necrotic Fungal Phytopathogen Botrytis cinerea

2014 ◽  
Vol 27 (8) ◽  
pp. 781-792 ◽  
Author(s):  
Majse Nafisi ◽  
Maria Stranne ◽  
Lisha Zhang ◽  
Jan A. L. van Kan ◽  
Yumiko Sakuragi
Keyword(s):  
Cell Wall ◽  
Botrytis Cinerea ◽  
Plant Cell ◽  
Plant Pathogens ◽  
Plant Cell Wall ◽  
High Efficiency ◽  
Microbial Infection ◽  
Cell Wall Degrading Enzymes ◽  
Novel Host

The plant cell wall is one of the first physical interfaces encountered by plant pathogens and consists of polysaccharides, of which arabinan is an important constituent. During infection, the necrotrophic plant pathogen Botrytis cinerea secretes a cocktail of plant cell-wall-degrading enzymes, including endo-arabinanase activity, which carries out the breakdown of arabinan. The roles of arabinan and endo-arabinanases during microbial infection were thus far elusive. In this study, the gene Bcara1 encoding for a novel α-1,5-L-endo-arabinanase was identified and the heterologously expressed BcAra1 protein was shown to hydrolyze linear arabinan with high efficiency whereas little or no activity was observed against the other oligo- and polysaccharides tested. The Bcara1 knockout mutants displayed reduced arabinanase activity in vitro and severe retardation in secondary lesion formation during infection of Arabidopsis leaves. These results indicate that BcAra1 is a novel endo-arabinanase and plays an important role during the infection of Arabidopsis. Interestingly, the level of Bcara1 transcript was considerably lower during the infection of Nicotiana benthamiana compared with Arabidopsis and, consequently, the ΔBcara1 mutants showed the wild-type level of virulence on N. benthamiana leaves. These results support the conclusion that the expression of Bcara1 is host dependent and is a key determinant of the disease outcome.

Plant cell wall reinforcement in the disease-resistance response: molecular composition and regulation

1995 ◽  
Vol 73 (S1) ◽  
pp. 511-517 ◽  
Author(s):  
Ulrich Matern ◽  
Bernhard Grimmig ◽  
Richard Edward Kneusel
Keyword(s):  
Cell Wall ◽  
Disease Resistance ◽  
Plant Cell ◽  
Plant Cell Wall ◽  
De Novo ◽  
Alternative Pathway ◽  
Petroselinum Crispum ◽  
Phenylpropanoid Metabolism ◽  
Resistance Response ◽  
Cell Wall Reinforcement

The disease-resistance response of plant cells is composed of a multitude of biochemical events, and the activation of one of these, the phenylpropanoid metabolism, is pivotal for the survival of cells under stress conditions. The basic features of this facet of the disease-resistance response are beginning to be unraveled in model plant cell culture systems. These studies revealed a novel, alternative pathway for the synthesis of cell wall bound hydroxycinnamoyl esters and lignin. The investigations have, therefore, set the stage for a detailed analysis of the induction process that includes fast, posttranslational activation mechanisms as well as de novo enzyme synthesis. The biosynthesis of phenolic compounds destined for the cell wall is considered to reach far beyond the mere physical strengthening of the cells and includes additional functions, e.g., the release of antimycotic hydroxybenzaldehydes, which are vital for stress compensation. Key words: elicitor-induced phenylpropanoids, cell wall reinforcement, hydroxycinnamoyl esters, lignin, caffeoyl-CoA-specific 3-O-methyltransferase, disease resistance response, parsley (Petroselinum crispum) cell cultures.

Ultrasound Assisted Solid-Liquid Extraction Devices - Harnessing Active Ingredients from Plant materials – Overview and Analysis

2020 ◽  
Vol 13 ◽  
Author(s):  
Venkatasubramanian Sivakumar
Keyword(s):  
Mass Transfer ◽  
Cell Wall ◽  
Plant Cell ◽  
Plant Cell Wall ◽  
Liquid Extraction ◽  
Active Ingredients ◽  
Plant Materials ◽  
Ultrasound Assisted ◽  
Solid Liquid Extraction ◽  
Solid Liquid

Background: In the growing environmental concern use of natural products, efficient processes and devices are necessary. Solid-Liquid extraction of active Ingredients from Plant materials is one of the important unit operations in Chemical Engineering and need to be enhanced. Objectives: Since, these active ingredients are firmly bound to the plant cell wall membrane, which pose mass-transfer resistance and need to get detached through the use of suitable process intensification tools such as ultrasound and suitable devices. Therefore, detailed analysis and review is essential on development made in this area through Publications and Patents. Hence, the present paper illustrates the development of ultrasound assisted device for solid-liquid extraction are presented in this paper. Methods: Advantages such as % Yield, Reduction in extraction time, use of ambient conditions, better process control, avoidance or minimizing multi stage extraction could be achieved due to the use of ultrasound in extraction as compared to conventional processes. Conclusions: Use of ultrasound to provide significant improvements in the extraction of Vegetable tannins, Natural dyes for application in Leather processing has been demonstrated and reported earlier. These enhancement could be possible through various effects of ultrasound such as better flow of solvents through micro-jet formation, mass transfer enhancement due to rupture of plant cell wall membranes through acoustic cavitation, better leaching due to micro-mixing and acoustic streaming effects. This approach would minimize material wastage; thereby, leading to eco-conservation of plant materials, which is very much essential for better environment. Hence, various methods and design for application of ultrasound assisted solid-liquid extractor device are necessary.

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