title: Effects of trehalose and sorbitol on the activity and structure of Pseudomonas cepacia lipase: spectroscopic insight.
abstract: The stability of enzymes with no reduction in their catalytic activity still remains a critical issue in industrial applications. Naturally occurring osmolytes are commonly used as protein stabilizers. In this study we have investigated the effects of sorbitol and trehalose on the structural stability and activity of Pseudomonas cepacia lipase (PCL), using UV-visible, circular dichroism (CD) and fluorescence spectroscopy. Surface plasmon resonance (SPR) technique was used to trace changes in the refractive index and dielectric constant of the environment. The results revealed that catalytic activity and intrinsic fluorescence intensity of PCL increased in the presence of both osmolytes. Far-UV CD spectra indicated that the protein has undergone some conformational changes upon interacting with these osmolytes. Increasing the concentration of sorbitol led to changes in the refractive index and consequently the dielectric constant of environment; whereas in the case of trehalose, such changes were not significant. Unfavorable interactions of trehalose with protein surface induced higher preferential exclusion from the enzyme-water interface than that of sorbitol. Results of this report could give further insights about the stabilization mechanism of osmolytes.
authors: Department of Biophysics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.
authors: Azadeh
authors: A
authors: Azizi
authors: Bijan
authors: B
authors: Ranjbar
authors: Khosro
authors: K
authors: Khajeh
authors: Tayebeh
authors: T
authors: Ghodselahi
authors: Soraya
authors: S
authors: Hoornam
authors: Hamid
authors: H
authors: Mobasheri
authors: Mohamad Reza
authors: MR
authors: Ganjalikhany
mesh_terms: D016956:Burkholderia cepacia / Q000201:enzymology*; D002942:Circular Dichroism; D006046:Gold / Q000737:chemistry; D006868:Hydrolysis / Q000187:drug effects; D008049:Lipase / Q000737:chemistry* / Q000378:metabolism*; D053768:Metal Nanoparticles / Q000737:chemistry; D013012:Sorbitol / Q000494:pharmacology*; D013050:Spectrometry, Fluorescence; D013329:Structure-Activity Relationship; D020349:Surface Plasmon Resonance; D014199:Trehalose / Q000494:pharmacology*
chemical_list: D013012:Sorbitol; D006046:Gold; D014199:Trehalose; D008049:Lipase