A novel multifunctional GH9 enzyme from Paenibacillus curdlanolyticus B-6 exhibiting endo/exo functions of cellulase, mannanase and xylanase activities
| Autor: | Vanarat Phakeenuya, Khanok Ratanakhanokchai, Akihiko Kosugi, Chakrit Tachaapaikoon |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Glycoside Hydrolases
Oligosaccharides Cellulase Cellobiose Applied Microbiology and Biotechnology Substrate Specificity 03 medical and health sciences chemistry.chemical_compound Hydrolysis Bacterial Proteins Polysaccharides Catalytic Domain Mannosidases Escherichia coli Glycoside hydrolase Enzyme kinetics Cellulose 030304 developmental biology 0303 health sciences biology 030306 microbiology food and beverages General Medicine Xylan Multifunctional Enzymes Recombinant Proteins Molecular Docking Simulation Kinetics Xylosidases chemistry Biochemistry Mutation biology.protein Xylanase Paenibacillus Biotechnology |
| Zdroj: | Applied microbiology and biotechnology. 104(5) |
| ISSN: | 1432-0614 |
| Popis: | PcMulGH9, a novel glycoside hydrolase family 9 (GH9) from Paenibacillus curdlanolyticus B-6, was successfully expressed in Escherichia coli. It is composed of a catalytic domain of GH9, two domains of carbohydrate-binding module family 3 (CBM3) and two domains of fibronectin type 3 (Fn3). The PcMulGH9 enzyme showed broad activity towards the β-1,4 glycosidic linkages of cellulose, mannan and xylan, including cellulose and xylan contained in lignocellulosic biomass, which is rarely found in GH9. The enzyme hydrolysed substrates with bifunctional endo-/exotypes cellulase, mannanase and xylanase activities, but predominantly exhibited exo-activities. This enzyme released cellobiose as a major product from cellohexaose, while mannotriose and xylotriose were major hydrolysis products from mannohexaose and xylohexaose, respectively. Moreover, PcMulGH9 could hydrolyse untreated corn hull and rice straw into xylo- and cello-oligosaccharides. Enzyme kinetics, site-directed mutagenesis and molecular docking revealed that Met394, located at the binding subsite + 2, was involved in broad substrate specificity of PcMulGH9 enzyme. This study offers new knowledge of the multifunctional cellulase/mannanase/xylanase in GH9. The PcMulGH9 enzyme showed a novel function of GH9, which increases its potential for saccharification of lignocellulosic biomass into value-added products, especially oligosaccharides. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full text from SpringerLink