Crystallization and preliminary X-ray analysis of Alicyclobacillus acidocaldarius endoglucanase CelA.
نویسندگان
چکیده
Crystallization of a family 9 beta-1,4-glucanase from the thermoacidophilic bacterium Alicyclobacillus acidocaldarius CelA is reported. Thin plates can be obtained by hanging-drop vapour-diffusion crystallization in high concentrations (60%) of MPD. These crystals are unusual in that they do not bind the dye IZIT in the mother liquor and do not appear to dissolve in water after three weeks or in the storage buffer after 2 d. The crystals diffract weakly and the diffraction pattern is compatible with crystal disorder in one direction. After testing several crystals at the ESRF beamlines ID14-1 and ID14-2, a crystal was found which gave ordered diffraction in all directions. A full data set was collected to 3.0 A resolution, which allowed unambiguous determination of the space group as P2(1)2(1)2 and the unit-cell parameters as a = 85, b = 129.7, c = 48.6 A. Initial promising results from molecular-replacement searches are reported.
منابع مشابه
A Comparative Study of Activity and Stability of the Free and the Immobilized Endoglucanase from Alicyclobacillus Acidocaldarius
AaCel9A [β-1,4-endoglucanase, (E.C:3.2.1.4)], was immobilized onto glutaraldehyde activated chitosan macrosphere by covalent attachment. The properties of the immobilized AaCel9A were investigated by determining the optimum pH and optimum temperature for activity, thermal stability, and kinetic parameters. The immobilization process shifted the enzyme’s optimum temperature from 65 °C for the fr...
متن کاملCrystallization and preliminary X-ray study of native and selenomethionyl beta-1,4-mannanase AaManA from Alicyclobacillus acidocaldariusTc-12-31.
AaManA, a beta-1,4-mannanase from the thermoacidophile Alicyclobacillus acidocaldarius Tc-12-31, was expressed in Escherichia coli and purified in a form suitable for X-ray crystallographic analysis. Crystals were obtained using the hanging-drop vapour-diffusion method at 291 K using ammonium dihydrogen phosphate as a precipitant. Data were collected from native mannanase and from a selenomethi...
متن کاملExpression, purification, crystallization and preliminary X-ray diffraction analysis of Aspergillus terreus endo-β-1,4-glucanase from glycoside hydrolase family 12.
Endoglucanases are important enzymes that are involved in the modification and degradation of cellulose. Filamentous fungi such as Aspergillus terreus are effective biomass degraders in nature owing to their capacity to produce an enzymatic arsenal of glycoside hydrolases, including endoglucanase from glycoside hydrolase family 12 (GH12). The A. terreus GH12 endoglucanase was cloned and overexp...
متن کاملA potential fortuitous binding of inhibitors of an inverting family GH9 β-glycosidase derived from isofagomine.
Using structural insight, the binding mode of isofagomine-derived inhibitors with family GH9 glycosidases is achieved via the study of Alicyclobacillus acidocaldarius (AaCel9A) endoglucanase. In contrast to what was observed in the first report using these compounds with inverting glycosidases from family GH6, these inhibitors do not adopt a distorted conformation in the active site.
متن کاملComplete genome sequence of Alicyclobacillus acidocaldarius strain Tc-4-1.
Alicyclobacillus acidocaldarius strain Tc-4-1 was initially isolated from a hot spring in Tengchong, China. This organism is both thermophilic and acidophilic. It can produce heat- and acid-stable enzymes, such as amylase and esterase, which may be important in industry. Here we report the whole genome sequence of the strain.
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Acta crystallographica. Section D, Biological crystallography
دوره 59 Pt 1 شماره
صفحات -
تاریخ انتشار 2003