Secondary structure, conformational stability and glycosylation of a recombinant Candida rugosa lipase studied by Fourier-transform infrared spectroscopy

The Biochemical Journal
Antonino NatalelloSilvia Maria Doglia

Abstract

The secondary structure of lipase 1 from Candida rugosa, a model system for large monomeric enzymes, has been studied by FTIR (Fourier-transform infrared) spectroscopy in water and 2H2O. The secondary structure content, determined by the analysis of the amide I band absorption through second derivative and curve fitting procedures, is in agreement with that estimated by X-ray data and predicts, in addition, the existence of two classes of alpha-helices. We have also investigated the enzyme stability and aggregation at high temperature by following the protein unfolding. The thermal stability determined by FTIR is in excellent agreement with the temperature dependence of the lipase activity. Furthermore, new insights on the glycosylation of the recombinant protein produced in Pichia pastoris and on its heterogeneity related to different fermentation batches were obtained by the analysis of the IR absorption in the 1200-900 cm(-1) carbohydrate region. A drastic reduction of the intensity of this band was found after enzymic deglycosylation of the protein. To confirm that the FTIR absorption in the 1200-900 cm(-1) region depends on the carbohydrate content and glycoform distribution, we performed an MS analysis of the protein suga...Continue Reading

References

Apr 1, 1992·Protein Engineering·D L OllisJ Schrag
Nov 21, 1995·Proceedings of the National Academy of Sciences of the United States of America·K Griebenow, A M Klibanov
Apr 27, 1995·Biochimica Et Biophysica Acta·J M HaddenD C Lee
Jan 1, 1994·Protein Science : a Publication of the Protein Society·P GrochulskiM Cygler
Jan 1, 1993·Progress in Biophysics and Molecular Biology·J L ArrondoF M Goñi
May 30, 1998·Protein Science : a Publication of the Protein Society·I EchabeJ L Arrondo
Jun 23, 1998·Quarterly Reviews of Biophysics·L K Tamm, S A Tatulian
Jul 9, 1998·Protein Science : a Publication of the Protein Society·S BroccaR D Schmid
Oct 3, 1999·Methods in Enzymology·S SeshadriA L Fink
Dec 22, 1999·Progress in Biophysics and Molecular Biology·J L Arrondo, F M Goñi
Jun 13, 2000·Protein Science : a Publication of the Protein Society·S BroccaM Lotti
Aug 10, 2000·European Journal of Biochemistry·C FiniP Pucci
Jul 3, 1998·Angewandte Chemie·Rolf D Schmid, Robert Verger

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Citations

Feb 2, 2013·Applied Microbiology and Biotechnology·Carlo SantambrogioMarina Lotti
Nov 7, 2012·Journal of Molecular Microbiology and Biotechnology·Jianhua ZhengQi Jin
Jan 9, 2009·Bioscience, Biotechnology, and Biochemistry·Takeshi FuruhashiGerhard Steiner
Jan 15, 2015·Biosensors·Valeria CalifanoLuciano R M Vicari
Jun 14, 2005·FEBS Letters·Diletta AmiSilvia Maria Doglia
Jan 23, 2016·International Journal of Biological Macromolecules·Mario RaspantiLaura Cipolla
May 12, 2012·Biochimica Et Biophysica Acta·C GruianV Simon
Apr 7, 2010·International Journal of Biological Macromolecules·Wael M ElshemeyAnwar A Elsayed
Nov 17, 2009·Bioresource Technology·Aikaterini A TziallaHaralambos Stamatis
Jul 30, 2008·Protein Expression and Purification·Gaetano InvernizziMarina Lotti
Mar 21, 2008·Biotechnology and Bioengineering·Nuria Gonzalez-MontalbanSilvia Maria Doglia
Nov 17, 2007·Journal of Peptide Science : an Official Publication of the European Peptide Society·K B Joshi, Sandeep Verma
Jan 26, 2006·Biochimica Et Biophysica Acta·Diletta AmiSilvia Maria Doglia
Nov 18, 2005·Biotechnology Advances·Pablo Domínguez de MaríaAndrés R Alcántara
Apr 4, 2015·Enzyme and Microbial Technology·Nur Royhaila MohamadRoswanira Abdul Wahab
Jan 30, 2008·Protein Expression and Purification·Antonino NatalelloArio de Marco
Jul 5, 2012·International Journal of Pharmaceutics·Nikola MilašinovićMelina Kalagasidis Krušić
Apr 1, 2017·Die Naturwissenschaften·Paulina BolibokArtur P Terzyk
Dec 4, 2014·Colloids and Surfaces. B, Biointerfaces·Adriano L SouzaOsvaldo N Oliveira
Nov 16, 2005·Proteins·Véronique Receveur-BréchotSonia Longhi
Oct 28, 2019·Preparative Biochemistry & Biotechnology·Sema SalgınUğur Salgın
Aug 6, 2020·Angewandte Chemie·Lei WangSamuel Sánchez
Dec 6, 2017·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Valeria CalifanoLuciano Vicari

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