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Characterization of fusing CBM6 within Penicillium oxalicum endo-1,4-beta-xylanase GH11 and its effectiveness on insoluble degradation |
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| รหัสดีโอไอ | |
| Title | Characterization of fusing CBM6 within Penicillium oxalicum endo-1,4-beta-xylanase GH11 and its effectiveness on insoluble degradation |
| Creator | Achara Chaiongkarn |
| Contributor | Jirapa Dathong, Nomchit Kaewthai Andrei, Patthanant Natpinit, Premsuda Saman, Chutima Kuancha |
| Publisher | The Thai Society for Biotechnology |
| Publication Year | 2562 |
| Keyword | CBM6, Penicillium oxalicum endo 1,4 beta xylanase GH11, Pichia pastoris |
| Abstract | Xylanase is rapidly gaining attention as suitable for applications in a wide variety of industrial processes such as food additives, animal feed, pulp bleaching and ethanol or xylitol production. Endo-1,4-beta-xylanase GH11 in Pichia pastoris expression, xyn11- CBM1 and xyn11- CBM1- CBM6 were constructed and characterized to investigate the influence of CBM6 on Penicillium oxalicum. Multiple copy genes of recombinant Pichia strains were selected on YPD agar + 2,000 ? g/ mL of ZeocinTM and xylan- Congo red clearance plate assay. Highest activities of Xyn11-CBM1 and Xyn11-CBM1CBM6 were 39.07? 0.79 and 47.48? 0.33 units/mg protein, respectively at optimal conditions (citrate buffer pH 5.0, at 45?C). Xyn11-CBM1 and Xyn11CBM1-CBM6 showed an acidic-stable property at pH 4.0-6.0 and 4.0-6.5, respectively ( more than 100% of activity remaining) when kept at room temperature (RT) for 240 minutes. Xyn11-CBM1-CBM6 exhibited more than 85% and 75% of activity remaining in phosphate buffer pH 6.0 at 45?C for 60 and 120 minutes, respectively and significantly higher than Xyn11-CBM1. The activity of both enzymes was strongly inhibited by 1 mM of Hg2+and showed over-estimated sizes (about 22.96 and 57.36 kDa) when separated by SDS- PAGE and zymogram analysis. Degradation of complex materials indicated that the highest reducing sugar release was produced by Xyn11 CBM1-CBM6 activity. TLC analysis revealed that xylobiose, xylotriose and xylotetraose were produced by both enzymes when they hydrolyzed corncob, corn leaves and stalks, and rice straw. Results indicated that Xyn11-CBM1CBM6 was a suitable candidate for application in various industries. |
| Language | EN |
| URL Website | http://tsb2019.com/ |
| Website title | The 31st Annual Meeting of the Thai Society for Biotechnology and International Conference (TSB 2019) |
