What is the role of glutamine residues outside the catalytic site in a Beta-glycosidase?

I'm working with a beta-glycosidase from a hyperthermophilic archaeon. Eventhough it is highly thermostable, its kcat values are very low. So we tried to mutate some catalytic site and non...

03 April 2015 145 7 View

How do I perform calculation of pseudo first order kinetic parameters from experimental data?

31 December 2014 1,850 4 View

How to calculate activation energy (Ea) from a non-linear Arrhenius plot?

I have to calculate the activation energy (Ea) for a thermostable enzyme (wild-type) and its mutants based on its enzyme activity (ln of activity vs 1/T). The wild-type is stable at near boiling...

01 February 2014 3,385 7 View

What are the tools available to predict disulfide connectivity in proteins?

I am looking for some tools to predict the cysteine bonding state and disulfide connectivity of a protein from its sequence.

03 April 2013 2,566 2 View

Can N-Terminal His-tag change the properties of the expressed recombinant protein?

We expressed a B-Glycosidase enzyme in Ecoli BL21 cells with a N-Terminal His-tag. The same enzyme has been expressed in BL21 previously but without His-tag. The enzyme with His-tag showed a...

02 March 2013 1,636 18 View

Is enzyme kinetics determined from discontinuous enzyme assay reliable?

To study the kinetics of an enzyme, continuous enzyme assay was the most preferred method. But in some cases, the discontinuous assay has been used. So how reliable are the kinetic data obtained...

09 October 2012 9,898 6 View

What is the action of Sodium Carbonate as stop solution in a discontinuous enzyme assay?

Sodium Carbonate is used as a stop solution in enzyme assays. How does it act on the enzyme substrate complex to stop the reaction?

07 August 2012 6,508 9 View

Procedure for Transglycosylation reaction of a hyperthermophilic Beta-Glycosidase

Can anyone suggest a method to study the transglycosylation reaction of Beta-Glycosidase from a hyperthermophilic organism. I prefer to use TLC to analyze the transglycosylated products.

07 August 2012 3,333 1 View

Molar extinction coefficient of 2-nitrophenol and 4-nitrophenol

I am doing Beta-Glucosidase assay from a hot spring archaea. The assay conditions include Sodium citrate buffer, pH 5.0, temperature at 70C and the substrates are 2-Nitrophenylglycoside and...

06 July 2012 2,738 5 View

Application of DSC for enzyme thermostability

I am characterizing an enzyme from hyperthermophilic organism. For measuring the thermostability of the enzyme, what are the methods available? Is using Differential Scanning Calorimetry (DSC)...

05 June 2012 7,665 11 View

How to calculate enzyme's energy of activation without rate constant (+excel)?

I want to calculate an enzyme's energy of activation, but I don't have the enzyme's molecular weight to calculate the rate constant or the pre-exponential factor A. This is part of an assignement...

25 December 2020 8,738 3 View

What is apparent Km Value of an Enzyme, Is it different from usual Km value and Why it is important?

It is very much confusing when some papers use this term "apparent Km of an enzyme even if they use single substrate ? Not at all clear ! Please help me understanding this. Thanks in advance.

27 September 2020 3,522 7 View

What is the difference between PCR and the initial steps (before SDS gel) of DNA sequencing? Which method is more effective?

I apologize if my question is not clear. I have a pure chemistry background and I am very new in a Protein Chemistry and Enzymology science.

25 September 2020 7,968 5 View

How to isolate/quantify a fungal enzyme for comparison of control vs. treatment?

I want to compare the amount of an enzyme's expression (laccase for example) or presence in the tissue as a result of the experimental treatment. Is there a protein similar to an antibody that...

31 December 2019 1,359 2 View

How strong is Flag tag binding to Anti-Flag antibody?

I am trying to plan an pulldown assay to find some prey protein for my bait protein. I am interested in using the flag tag because it is small and doesn't disturb structure and doesn't require...

11 December 2019 2,406 3 View

How to calculate the dosage of enzymes in wheat flour?

I have recently started working on improvement of wheat flour for baking. I have come across a research which involved the usage of xylanase, cellulase, beta-glucanase and carbohydrases for...

10 November 2019 3,608 1 View

Can I restrict the amount of one species when using KinTek for kinetic fitting?

I am using KinTek software to do a kinetic fitting. I have determined kinetics of the formations of several species in the time course of 5min and established a model for the enzyme (just as an...

09 October 2019 3,835 4 View

I am searching the pGex or pET plazid of HDAC6, if he is possibly for somebody and would give for me. Does somebody have this plasmid?

I am searching the pGex or pET plazid of HDAC6, if he is possibly for somebody and would give for me, would be needed for my work. I would like to test the interaction between my labor's new...

05 June 2019 4,133 1 View

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08 January 2019 3,512 5 View

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31 December 2018 8,083 5 View