PMID: 11427942Jun 28, 2001

Novel kinetic analysis of enzymatic dipeptide synthesis: effect of pH and substrates on thermolysin catalysis

Biotechnology and Bioengineering
Y MurakamiA Hirata


The point of maximum activity is specific to a particular substrate-enzyme system but may vary with different substrates and the same enzyme. The specificity of enzymes has, however, been generally reported only at their "optimal" pH. In this article, we introduce the Michaelis-Menten equation taking pH into account, and apply it to the pH-activity profile of the thermolysin-catalyzed dipeptide synthesis. It has been reported to date that the pH-activity profile of thermolysin follows a bell-shaped curve with a maximal activity at or near pH 7.0. The profiles obtained in this study, however, indicated that the optimal pH varied from 5.8 (for F-AspPheOMe) to 7.3 (for Z-ArgPheOMe), and the order of thermolysin activity was greatly dependent on the pH of reaction media. We have succeeded in evaluating the substrates-induced change of the dissociation states of the active site of thermolysin using the hydrophobicity of substrates. We have obtained apparent kinetic parameters which are independent of the pH of reaction media. The apparent specificity of thermolysin which were independent of pH of the reaction media was in order L-Leu > L-Asp > L-Arg > L-Ala > L-Gly > L-Val and Z > Boc = F at P1 and P2 positions, respectively.


Feb 12, 1970·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·A Berger, I Schechter
Mar 27, 1968·Biochemical and Biophysical Research Communications·K Morihara, T Oka
Mar 11, 1968·Archives of Biochemistry and Biophysics·K MoriharaT Oka
Jul 26, 1968·Biochemical and Biophysical Research Communications·J Feder
Aug 1, 1970·European Journal of Biochemistry·K Morihara, H Tsuzuki
Sep 1, 1971·Archives of Biochemistry and Biophysics·K Morihara, H Tsuzuki
Dec 1, 1971·Archives of Biochemistry and Biophysics·C E Stauffer
Mar 13, 1981·Biochimica Et Biophysica Acta·K MartinekI V Berezin
Jun 7, 1982·FEBS Letters·M PankA Aaviksaar
Oct 5, 1982·Journal of Molecular Biology·M A Holmes, B W Matthews
Sep 1, 1980·Journal of Biochemistry·T Oka, K Morihara
Sep 26, 1995·Biochemistry·P P WangikarJ S Dordick
Jan 4, 1997·Biochimica Et Biophysica Acta·T LignéV Larreta-Garde
Jul 1, 1997·Journal of Biochemistry·S KawaguchiS Kuramitsu
May 1, 1994·Biotechnology and Bioengineering·T NagayasuK Nakanishi
May 1, 1994·Biotechnology and Bioengineering·T NagayasuK Nakanishi
Jun 20, 1995·Biotechnology and Bioengineering·M MiyanagaK Nakanishi


Related Concepts

Carboxylic Acids
Hydrogen-Ion Concentration
Substrate Specificity

Trending Feeds


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.

Synthetic Genetic Array Analysis

Synthetic genetic arrays allow the systematic examination of genetic interactions. Here is the latest research focusing on synthetic genetic arrays and their analyses.

Congenital Hyperinsulinism

Congenital hyperinsulinism is caused by genetic mutations resulting in excess insulin secretion from beta cells of the pancreas. Here is the latest research.

Neural Activity: Imaging

Imaging of neural activity in vivo has developed rapidly recently with the advancement of fluorescence microscopy, including new applications using miniaturized microscopes (miniscopes). This feed follows the progress in this growing field.

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.

Epigenetic Memory

Epigenetic memory refers to the heritable genetic changes that are not explained by the DNA sequence. Find the latest research on epigenetic memory here.

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.

Femoral Neoplasms

Femoral Neoplasms are bone tumors that arise in the femur. Discover the latest research on femoral neoplasms 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.