- Title
- Delineating functional properties of a cello-oligosaccharide and Bglucan specific cellobiohydrolase (GH5_38): Its synergism with Cel6A and Cel7A for B-(1,3)-(1,4)-glucan degradation
- Creator
- Mafa, Mpho S, Malgas, Samkelo, Rashamuse, Konanani, Pletschke, Brett I
- Subject
- To be catalogued
- Date
- 2020
- Type
- text
- Type
- article
- Identifier
- http://hdl.handle.net/10962/429425
- Identifier
- vital:72609
- Identifier
- xlink:href="https://doi.org/10.1016/j.carres.2020.108081"
- Description
- Cellulase cocktails formulated to degrade crystalline cellulose generally contain cellobiohydrolases (CBHs), referred to as CBHI (Cel7A) and CBHII (Cel6A), as the major constituents. The combined hydrolytic activities of CBHI and CBHII improve the release of fermentable sugars (β-1,4-cellobiose as the main product) from crystalline cellulose. In this study, a novel cellobiohydrolase (Exg-D) sourced from a metagenome of hindgut bacterial symbionts of a termite was heterologouly expressed, purified, and functionally characterised. Exg-D specific activity was higher on insoluble barley β-glucan (38.94 U/mg protein), soluble wheat flour β-glucan (12.71 U/mg protein) and oat β-glucan (8.89 U/mg protein) compared to cellulosic substrates; Avicel and CMC. We further explored Exg-D activity on the unpretreated or NaOH-pretreated (mercerised) Avicel and compared its activity to commercially available CBHI and CBHII on these celluloses. CBHI displayed the highest activity of 4.74 U/mg protein on mercerised cellulose followed by CBHII (2.14 U/mg protein), while Exg-D activity on untreated and mercerised cellulose was 1.66 and 1.67 U/mg protein, respectively. The high activity of CBHI was supported by binding assays, which revealed that CBHI has a higher binding capacity towards crystalline cellulose compared to Exg-D and CBHII. Only CBHI and CBHII showed synergism during the hydrolysis of mercerised Avicel, showing a degree of synergy (DS) of about 1.299 and yielded about 1.43 μmol/ml of reducing sugars higher than control. In contrast, Exg-D and CBHII displayed synergism during β-glucan degradation, displaying a DS of about 1.22. Thus, we propose that Exg-D should only be used synergistically with other CBHs to degrade mixed linked-β-(1,3)-(1,4)-glucan.
- Format
- computer, online resource, application/pdf, 1 online resource (8 pages), pdf
- Publisher
- Elsevier
- Language
- English
- Relation
- Carbohydrate Research, Mafa, M.S., Malgas, S., Rashamuse, K. and Pletschke, B.I. (2020) Delineating functional properties of a cello-oligosaccharide and Bglucan specific cellobiohydrolase (GH5_38): Its synergism with Cel6A and Cel7A for B-(1,3)-(1,4)-glucan degradation. Carbohydrate Research. 495 (2020). p.1-8, Carbohydrate Research volume 495 number 2020 p. 1 2020 1873-426X
- Rights
- Publisher
- Rights
- Use of this resource is governed by the terms and conditions of the Elsevier Terms and Conditions Statement (https://www.elsevier.com/legal/elsevier-website-terms-and-conditions)
- Rights
- Closed Access
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Collections
Pletschke, Brett (Prof)
RU Department of Biochemistry, Microbiology and Bioinformatics
SDG 03. Good Health and Well-Being
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|---|---|---|---|---|---|---|---|
| View Details Download | SOURCE1 | Delineating functional properties of a cello-oligosaccharide and Bglucan specific cellobiohydrolase (GH5_38).pdf | 1 MB | Adobe Acrobat PDF | View Details Download |