1
vote
0answers
15 views

Enzyme Assay - pectinase

During assaying an enzyme at high temperature, the substrate (Pectin) is degraded by the high temperature rather than by enzyme, so, how can I minimize degradation of the substrate by the temperature? ...
2
votes
1answer
49 views

Pectinase Enzyme Assay

I am working on pectinase enzyme assay. I incubated 900 ul of substrate for 10 minutes in the water bath, followed by adding 2ml of DNSA reagent, then 100ul of enzyme extract added finally i read the ...
4
votes
1answer
63 views

DpnI over-digestion

We have a long protocol that we are optimizing that includes DpnI digestion of a PCR product (to remove any of the template DNA if it's methylated, and while we're not certain in the blind tests, ...
2
votes
1answer
37 views

What does “thermodynamic equilibrium” mean for an enzyme-substrate complex?

From Fersht, Enzyme Structure and Mechanism p. 87: The Michaelis-Menten mechanism assumes that the enzyme-substrate complex is in thermodynamic equilibrium with free enzyme and substrate. In ...
1
vote
1answer
40 views

Enzyme kinetics [closed]

I can't understand how to study enzyme kinetics. Say I have a lipase and want to study the kinetics of this lipase using a fluorogenic substrate, how would I do this? From what I understand I would ...
2
votes
1answer
34 views

Activity of glucokinase

From Solomon et al, 2013 ACC Synthetic biology and from this video : Here, there are 2 competing reactions for glucose - one with glk as enzyme and other with gdh as enzyme. In the graph, y axis ...
3
votes
3answers
119 views

Are there any methods to quantify H2O2 (hydrogen peroxide) which don't rely on horseradish peroxidase?

I can't use HRP since my substrates interfere with it and I need a real-time method because I want to measure kinetic parameters of some oxidases.
4
votes
2answers
320 views

Why are enzyme-catalysed reactions slower at lower substrate concentration?

Suppose I'm using 200 nmoles of enzyme and 2 mmoles of substrate. The enzyme should be saturated but if I use 50 mmoles of substrate, the reaction will be faster. Why? I just can't get it! Even at ...