title: Counteraction of Trehalose on N, N-Dimethylformamide-Induced Candida rugosa Lipase Denaturation: Spectroscopic Insight and Molecular Dynamic Simulation.
abstract: Candida rugosa lipase (CRL) has been widely used as a biocatalyst for non-aqueous synthesis in biotechnological applications, which, however, often suffers significant loss of activity in organic solvent. Experimental results show that trehalose could actively counteract the organic-solvent-induced protein denaturation, while the molecular mechanisms still don't unclear. Herein, CRL was used as a model enzyme to explore the effects of trehalose on the retention of enzymatic activity upon incubation in N,N-dimethylformamide (DMF). Results showed that both catalytic activity and conformation changes of CRL influenced by DMF solvent were inhibited by trehalose in a dose-dependent fashion. The simulations further indicated that the CRL protein unfolded in binary DMF solution, but retained the native state in the ternary DMF/trehalose system. Trehalose as the second osmolyte added into binary DMF solution decreased DMF-CRL hydrogen bonds efficiently, whereas increased the intermolecular hydrogen bondings between DMF and trehalose. Thus, the origin of its denaturing effects of DMF on protein is thought to be due to the preferential exclusion of trehalose as well as the intermolecular hydrogen bondings between trehalose and DMF. These findings suggest that trehalose protect the CRL protein from DMF-induced unfolding via both indirect and direct interactions.
authors.affiliation: College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 210009, PR China.
authors.forename: Xin
authors.initials: X
authors.lastname: Yang
authors.affiliation: College of Food Science and Light Industry, Nanjing Tech University, Nanjing 210009, PR China.
authors.forename: Ling
authors.initials: L
authors.lastname: Jiang
authors.affiliation: College of Food Science and Light Industry, Nanjing Tech University, Nanjing 210009, PR China.
authors.forename: Yigang
authors.initials: Y
authors.lastname: Jia
authors.affiliation: College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 210009, PR China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Hu
authors.affiliation: College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 210009, PR China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Xu
authors.affiliation: College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 210009, PR China.
authors.forename: Xian
authors.initials: X
authors.lastname: Xu
authors.affiliation: College of Pharmaceutical Sciences, Nanjing Tech University, Nanjing 210009, PR China.
authors.forename: He
authors.initials: H
authors.lastname: Huang
mesh_terms: D002175:Candida / Q000737:chemistry / Q000201:enzymology*; D004126:Dimethylformamide / Q000378:metabolism*; D005079:Excipients / Q000378:metabolism*; D007218:Industrial Microbiology; D008049:Lipase / Q000737:chemistry / Q000378:metabolism*; D056004:Molecular Dynamics Simulation; D011487:Protein Conformation / Q000187:drug effects; D011489:Protein Denaturation / Q000187:drug effects*; D014199:Trehalose / Q000378:metabolism*
chemical_list: D005079:Excipients; D004126:Dimethylformamide; D014199:Trehalose; D008049:Lipase