Engineering the pH-optimum of a triglyceride lipase: from predictions based on electrostatic computations to experimental results

Journal of Biotechnology
Maria Teresa Neves-PetersenSteffen B Petersen

Abstract

The optimisation of enzymes for particular purposes or conditions remains an important target in virtually all protein engineering endeavours. Here, we present a successful strategy for altering the pH-optimum of the triglyceride lipase cutinase from Fusarium solani pisi. The computed electrostatic pH-dependent potentials in the active site environment are correlated with the experimentally observed enzymatic activities. At pH-optimum a distinct negative potential is present in all the lipases and esterases that we studied so far. This has prompted us to propose the "The Electrostatic Catapult Model" as a model for product release after cleavage of the ester bond. The origin of the negative potential is associated with the titration status of specific residues in the vicinity of the active site cleft. In the case of cutinase, the role of Glu44 was systematically investigated by mutations into Ala and Lys. Also, the neighbouring Thr45 was mutated into Proline, with the aim of shifting the spatial location of Glu44. All the charge mutants displayed altered titration behaviour of active site electrostatic potentials. Typically, the substitution of the residue Glu44 pushes the onset of the active site negative potential towards mor...Continue Reading

References

Dec 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·F SangerA R Coulson
Apr 1, 1992·Proteins·A L MorrisJ M Thornton
Jan 1, 1992·Analytical Biochemistry·W P Deng, J A Nickoloff
Jul 1, 1990·Protein Engineering·D Mantafounis, J Pitts
Jan 1, 1989·Proteins·P CarterJ A Wells
Apr 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·C T ChungR H Miller
Dec 26, 1989·Biochemistry·K SomanR Fletterick
Oct 13, 1988·Nature·S LinseS Forsén
Jan 1, 1988·Proteins·M K Gilson, B H Honig
Jan 1, 1986·Progress in Biophysics and Molecular Biology·N K Rogers
Mar 1, 1987·Proceedings of the National Academy of Sciences of the United States of America·J A WellsD A Estell
Jan 1, 1985·CRC Critical Reviews in Biochemistry·J B MatthewS J Shire
Aug 5, 1985·Journal of Molecular Biology·M K GilsonB Honig
May 23, 1972·Biochemistry·C Tanford, R Roxby
Aug 1, 1984·Quarterly Reviews of Biophysics·A Warshel, S T Russell
Aug 7, 1983·Journal of Theoretical Biology·J Angyán, G Náray-Szabó
Jan 7, 1982·Nature·J M Thornton
Sep 28, 1982·Biochemistry·J B Matthew, F M Richards
Apr 1, 1995·Current Opinion in Structural Biology·M K Gilson
May 26, 1995·Science·B Honig, A Nicholls
Aug 31, 1994·Journal of Biotechnology·H W AnthonsenS B Petersen
Jan 11, 1994·Biochemistry·C MartinezC Cambillau

❮ Previous
Next ❯

Citations

May 17, 2007·Biotechnology Letters·Jacob Dam SørensenMarilyn G Wiebe
May 17, 2011·Physical Biology·Zhe ZhangEmil Alexov
Feb 13, 2010·Protein Engineering, Design & Selection : PEDS·Damien Yann ColinBrigitte Kerfelec
Feb 16, 2002·Protein Science : a Publication of the Protein Society·Maria Teresa Neves-PetersenSteffen Bjørn Petersen
Jul 15, 2011·Clinical and Experimental Gastroenterology·Aaron FiekerMartine Armand
May 2, 2014·BioMed Research International·Cristiane D AnobomDenise M Freire
Dec 14, 2004·Chemistry and Physics of Lipids·Torben SnabeSteffen Bjørn Petersen
Dec 14, 2007·Journal of Biotechnology·Ji Hye YangYoung Je Yoo
Oct 21, 2004·Critical Reviews in Biotechnology·Sanjeev K AhujaAntonio R Moreira
Sep 2, 2018·International Journal of Biological Macromolecules·Osmar Soares da SilvaTatiana Souza Porto

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.

Related Papers

Journal of Biotechnology
M T Neves PetersenSteffen B Petersen
Journal of Biotechnology
Steffen B PetersenE Hough
Journal of Biomedicine & Biotechnology
Steffen B PetersenMaria Teresa Neves Petersen
© 2021 Meta ULC. All rights reserved