Comparison of the kinetic specificity of subtilisin and thiolsubtilisin toward n-alkyl p-nitrophenyl esters

Biochemistry
M PhilippM L Bender

Abstract

The p-nitrophenyl esters of straight-chain fatty acids were used as substrates of the enzyme subtilisin Novo (EC 3.4.4.16) and its chemically produced artificial enzyme thiolsubtilisin. Subtilisin and thiolsubtilisin pH--activity profiles were determined, and kinetic effects of the active site O-S substitution were observed. Among the substrates tested, both enzymes show highest specificity with p-nitrophenyl butyrate. It was also found that subtilisin is more sensitive to changes in substrate chain length than is thiolsubtilisin. Second-order acylation rate constants (k2/Ks) are remarkably similar for both enzymes. However, thiolsubtilisin deacylation rate constants and Km values are lower than analogous subtilisin constants. While thiolsubtilisin deacylation rate constants give a pH profile identical with that of subtilisin, the pH profile of thiolsubtilisin acylation rate constants shows an active site pK value lowered from the subtilisin pK of 7.15 and exhibits an inflection point at pH 8.45, which is absent in subtilisin.

References

Jan 1, 1983·Molecular and Cellular Biochemistry·M Philipp, M L Bender
Jan 1, 1980·Advances in Enzyme Regulation·C C FanF M Huennekens
Apr 26, 2005·Biochimica Et Biophysica Acta·Aleksandra Djurdjevic-PahlJ Paul G Malthouse
Jan 11, 2001·Biochimica Et Biophysica Acta·P N Bryan
Jul 8, 2005·Applied and Environmental Microbiology·Carolin GöddeGarabed Antranikian
Jun 1, 1981·The Journal of Clinical Investigation·S G Gordon, B A Cross
Apr 4, 2007·Proceedings of the National Academy of Sciences of the United States of America·Ngolela Esther BabadyGrazia Isaya
Jun 14, 2002·FEBS Letters·Dar Fu Tai, Wen Chen Liaw
Oct 12, 1981·FEBS Letters·M Philipp, S Maripuri
Nov 15, 1992·Archives of Biochemistry and Biophysics·N Mukerjee, R Pietruszko
Nov 28, 1989·Biochemistry·J N HigakiC S Craik

Citations

Mar 7, 1973·Journal of the American Chemical Society·M L Bender, M Phillip
Nov 15, 1973·European Journal of Biochemistry·L Polgar, P Halász
Jun 15, 1974·FEBS Letters·M Philipp, M L Bender
Feb 9, 1972·Journal of the American Chemical Society·E C Lucas, M Caplow
Nov 1, 1966·Proceedings of the National Academy of Sciences of the United States of America·K E Neet, D E Koshland
Apr 1, 1970·Proceedings of the National Academy of Sciences of the United States of America·F C WedlerM L Bender
Dec 1, 1969·Proceedings of the National Academy of Sciences of the United States of America·L Polgar, M L Bender
Aug 1, 1969·Biochemistry·P M Enriquez, J T Gerig
Sep 1, 1967·Biochemistry·T H Fife, J B Milstien
Jan 1, 1965·Annual Review of Biochemistry·M L Bender, J KEZDY

Related Concepts

Enzymes, antithrombotic
Subtilisins
Subtilisin
Substrate Specificity
Enzymes for Treatment of Wounds and Ulcers
Hydrolase
Subtilisin BPN'
Chemicals
Subtilisin 72
PH Profile Measurement

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.

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.

Synapse Loss as Therapeutic Target in MS

As we age, the number of synapses present in the human brain starts to decline, but in neurodegenerative diseases this occurs at an accelerated rate. In MS, it has been shown that there is a reduction in synaptic density, which presents a potential target for treatment. Here is the latest research on synapse loss as a therapeutic target in MS.

Artificial Intelligence in Cardiac Imaging

Artificial intelligence (ai) techniques are increasingly applied to cardiovascular (cv) medicine in cardiac imaging analysis. 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.

Social Learning

Social learning involves learning new behaviors through observation, imitation and modeling. Follow this feed to stay up to date on the latest research.

Cell Atlas of the Human Eye

Constructing a cell atlas of the human eye will require transcriptomic and histologic analysis over the lifespan. This understanding will aid in the study of development and disease. Find the latest research pertaining to the Cell Atlas of the Human Eye here.

Single Cell Chromatin Profiling

Techniques like ATAC-seq and CUT&Tag have the potential to allow single cell profiling of chromatin accessibility, histones, and TFs. This will provide novel insight into cellular heterogeneity and cell states. Discover the latest research on single cell chromatin profiling here.

Genetic Screens in iPSC-derived Brain Cells

Genetic screening is a critical tool that can be employed to define and understand gene function and interaction. This feed focuses on genetic screens conducted using induced pluripotent stem cell (iPSC)-derived brain cells.