PMID: 11606210Oct 19, 2001Paper

Artificial chaperone mediated refolding of xylanase from an alkalophilic thermophilic Bacillus sp. Implications for in vitro protein renaturation via a folding intermediate

European Journal of Biochemistry
Devyani Nath, M Rao

Abstract

To gain insight into the molecular aspects of unfolding/refolding of enzymes from extremophilic organisms, we have used xylanase from an alkalophilic thermophilic Bacillus as the model system. Kinetics of denaturation/renaturation were monitored using intrinsic fluorescence studies. The protein fluorescence measurements suggested a putative intermediate state present in 0.08 M guanidine hydrochloride with an emission maximum of 345 nm; the far-UV circular dichroism spectra revealed content of secondary structure similar to the native enzyme. Studies with the fluorescent apolar probe 1-anilinonapthalene-8-sulfonate (1,8-ANS) were consistent with the presence of increased hydrophobic surfaces as compared with the native or fully unfolded protein. The refolding of Xyl II, was attempted by a relatively new strategy using an artificial chaperone assisted two-step method. The unfolded xylanase was found to bind to the detergent transiently and the subsequent addition of methyl-beta-cyclodextrin helped to strip the detergent and assist in the folding. Our findings suggested that the detergent stabilized a putative intermediate in the folding pathway seemingly equivalent to the folding state described as molten globule. The reactivatio...Continue Reading

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Citations

Sep 4, 2009·Bioprocess and Biosystems Engineering·Etsushi YamamotoTeruyuki Nagamune
Oct 8, 2005·Journal of Structural and Functional Genomics·Natalia OganesyanRosalind Kim
Jan 17, 2007·Biochemistry. Biokhimii︠a︡·Razieh YazdanparastReza Khodarahmi
Sep 4, 2004·Microbial Cell Factories·Luis Felipe Vallejo, Ursula Rinas
Mar 15, 2011·Biochimica Et Biophysica Acta·Daniel Otzen
Apr 19, 2015·Microbial Cell Factories·Anupam SinghAmulya Kumar Panda
Aug 14, 2012·International Journal of Biological Macromolecules·Yong-Qiang FanYu-Long Wang
Jun 3, 2006·Current Opinion in Biotechnology·Wieslaw Swietnicki
Apr 1, 2017·International Journal of Biological Macromolecules·Nuzhat GullRizwan Hassan Khan
Mar 22, 2014·Protein Science : a Publication of the Protein Society·Zhengrong YangChristie G Brouillette
Sep 12, 2012·Biotechnology Journal·Satoshi YamaguchiTeruyuki Nagamune
May 3, 2005·International Journal of Biological Macromolecules·Razieh YazdanparastReza Khodarahmi
Dec 1, 2005·Analytical Biochemistry·Hiroyuki ChikuKengo Sakaguchi
Aug 1, 2006·International Journal of Biological Macromolecules·Razieh YazdanparastFariba Khodagholi

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