title: Increases thermal stability and cellulose-binding capacity of Cryptococcus sp. S-2 lipase by fusion of cellulose binding domain derived from Trichoderma reesei.
abstract: To improve the thermal stability and cellulose-binding capacity of Cryptococcus sp. S-2 lipase (CSLP), the cellulose-binding domain originates from Trichoderma reesei cellobiohydrolase I was engineered into C-terminal region of the CSLP (CSLP-CBD). The CSLP and CSLP-CBD were successfully expressed in the Pichia pastoris using the strong methanol inducible alcohol oxidase 1 (AOX1) promoter and the secretion signal sequence from Saccharomyces cerevisiae (α factor). The recombinant CSLP and CSLP-CBD were secreted into culture medium and estimated by SDS-PAGE to be 22 and 27 kDa, respectively. The fusion enzyme was stable at 80 °C and retained more than 80% of its activity after 120-min incubation at this temperature. Our results also found that the fusion of fungal exoglucanase cellulose-binding domain to CSLP is responsible for cellulose-binding capacity. This attribute should make it an attractive applicant for enzyme immobilization.
authors.affiliation: Department of Biological Science, Faculty of Science, Ubon-Ratchathani University, Warinchumrab, Ubon-Ratchathani 34190, Thailand. jantaporn_25@yahoo.com
authors.forename: Jantaporn
authors.initials: J
authors.lastname: Thongekkaew
authors.forename: Hiroko
authors.initials: H
authors.lastname: Ikeda
authors.forename: Haruyuki
authors.initials: H
authors.lastname: Iefuji
mesh_terms: D002482:Cellulose / Q000737:chemistry*; D043366:Cellulose 1,4-beta-Cellobiosidase / Q000737:chemistry* / Q000235:genetics; D003454:Cryptococcus / Q000201:enzymology*; D004795:Enzyme Stability; D005656:Fungal Proteins / Q000737:chemistry* / Q000235:genetics; D006358:Hot Temperature; D008049:Lipase / Q000737:chemistry* / Q000235:genetics; D015202:Protein Engineering; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000235:genetics; D014242:Trichoderma / Q000201:enzymology
chemical_list: D005656:Fungal Proteins; D011993:Recombinant Fusion Proteins; D002482:Cellulose; D008049:Lipase; D043366:Cellulose 1,4-beta-Cellobiosidase