title: N-glycosylation affects the proper folding, enzymatic characteristics and production of a fungal ß-glucosidase.
abstract: Heterologous expression of ß-glucosidase is one of the approaches to enhance the efficiency of fungal cellulase preparations. It has been reported that N-glycosylation affects the structure framework, function and stability of proteins. In this study, a ß-glucosidase from Aspergillus terreus (GenBank: XP_001216552, BglS) was heterologously expressed in Pichia pastoris and Trichoderma reesei. The four asparagine residues were all linked with high-mannose-type oligosaccharides in P. pastoris, whereas only N224 carried high-mannosetype glycan in T. reesei (the other three sites carried one N-acetylglucosamine). The long N-glycan chains on PpBglS weakened its substrate affinity, activity and thermostability. The moderate post-translational and post-secretory glycan modification in T. reesei makes it a suitable expression system for BglS. The N224 glycan played a critical role in BglS folding. The elucidation of the correlation between the different N-glycosylation patterns of BglS and their corresponding enzymatic characteristics is an important step towards improving the activity, thermostability and even production of heterologous ß-glucosidase by glycan engineering.
authors.forename: Wei
authors.initials: W
authors.lastname: Wei
authors.forename: Ling
authors.initials: L
authors.lastname: Chen
authors.forename: Gen
authors.initials: G
authors.lastname: Zou
authors.forename: Qianfu
authors.initials: Q
authors.lastname: Wang
authors.forename: Xing
authors.initials: X
authors.lastname: Yan
authors.forename: Jun
authors.initials: J
authors.lastname: Zhang
authors.forename: Chengshu
authors.initials: C
authors.lastname: Wang
authors.forename: Zhihua
authors.initials: Z
authors.lastname: Zhou
mesh_terms: D001230:Aspergillus / Q000201:enzymology* / Q000235:genetics; D006031:Glycosylation; D010843:Pichia / Q000235:genetics* / Q000378:metabolism*; D017510:Protein Folding; D055550:Protein Stability; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014242:Trichoderma / Q000235:genetics* / Q000378:metabolism*; D001617:beta-Glucosidase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins; D001617:beta-Glucosidase