title: Ectoine-mediated protection of enzyme from the effect of pH and temperature stress: a study using Bacillus halodurans xylanase as a model.
abstract: Compatible solutes are small, soluble organic compounds that have the ability to stabilise proteins against various stress conditions. In this study, the protective effect of ectoines against pH stress is examined using a recombinant xylanase from Bacillus halodurans as a model. Ectoines improved the enzyme stability at low (4.5 and 5.0) and high pH (11 and 12); stabilisation effect of hydroxyectoine was superior to that of ectoine and trehalose. In the presence of hydroxyectoine, residual activity (after 10 h heating at 50 °C) increased from about 45 to 86 % at pH 5 and from 33 to 89 % at pH 12. When the xylanase was incubated at 65 °C for 5 h with 50 mM hydroxyectoine at pH 10, about 40 % of the original activity was retained while no residual activity was detected in the absence of additives or in the presence of ectoine or trehalose. The xylanase activity was slightly stimulated in the presence of 25 mM ectoines and then gradually decreased with increase in ectoines concentration. The thermal unfolding of the enzyme in the presence of the compatible solutes showed a modest increase in denaturation temperature but a larger increase in calorimetric enthalpy.
authors.affiliation: Department of Biotechnology, Center for Chemistry and Chemical Engineering, Lund University, P.O. Box 124, 22100 Lund, Sweden.
authors.forename: Doan
authors.initials: D
authors.lastname: Van-Thuoc
authors.forename: Suhaila O
authors.initials: SO
authors.lastname: Hashim
authors.forename: Rajni
authors.initials: R
authors.lastname: Hatti-Kaul
authors.forename: Gashaw
authors.initials: G
authors.lastname: Mamo
mesh_terms: D000599:Amino Acids, Diamino / Q000737:chemistry*; D001407:Bacillus / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D043364:Endo-1,4-beta Xylanases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D013696:Temperature
chemical_list: D000599:Amino Acids, Diamino; D001426:Bacterial Proteins; C045628:ectoine; D043364:Endo-1,4-beta Xylanases