My research is to assess nitrogen use efficiency in different wheat genotypes by determining the activity of ammonium assimilation enzymes which are mentioned above. But after a lot of struggle I wont find the complete protocols for these enzymes neither for root assay or shoot assay.
Kindly help me in this regard
waiting for positive response
The Brenda database for enzymes is an excellent resource. Look up each enzyme and you will see extensive data, including assays and substrates, and links to the publications that will describe the methods you are looking for.
https://www.brenda-enzymes.org
Dear Dr. Fatima Khan ,
There are protocols for the assays. I will share with you from the book on Practical Enzymology by Prof. Dr. Hans Bisswanger.
Regards,
Egbuna
Glutamine Synthetase EC 6.3.1.2
SN: l-glutamate:ammonia ligase (ADP-forming)
L-glutamate + ATP + NH3 → L-glutamine + ADP + Pi
Enzyme from Escherichia coli: dodecamer (12×51 800), Km (mM): 3.3 (l-glutamate),
0.2 (ATP), pH optimum 8.0. The assay is coupled with pyruvate kinase (PK) and
lactate dehydrogenase (LDH)
ADP + phosphoenolpyruvate
PK pyruvate + ATP
pyruvate + NADH + H+ LDH L-lactate + NAD+
For nitrate reductase, I recommend Analytical Techniques in Biochemistry
and Molecular Biology by Dr. Rajan Katoch.
The previous two assays are also described.
Regards,
Egbuna
Thankyou Mr Craig A Gelfand and Mr Chukwuebuka Egbuna for this brief guideline
- Badges
- Science topic
- Similar topics
- Microbiology
- Assays