Algae, Tree, Herbs, Bush, Shrub, Grasses, Vines, Fern, Moss, Spermatophyta, Bryophyta, Fern Ally, Flower, Photosynthesis, Eukaryote, Prokaryote, carbohydrate, vitamins, amino acids, botany, lipids, proteins, cell, cell wall, biotechnology, metabolities, enzymes, agriculture, horticulture, agronomy, bryology, plaleobotany, phytochemistry, enthnobotany, anatomy, ecology, plant breeding, ecology, genetics, chlorophyll, chloroplast, gymnosperms, sporophytes, spores, seed, pollination, pollen, agriculture, horticulture, taxanomy, fungi, molecular biology, biochemistry, bioinfomatics, microbiology, fertilizers, insecticides, pesticides, herbicides, plant growth regulators, medicinal plants, herbal medicines, chemistry, cytogenetics, bryology, ethnobotany, plant pathology, methodolgy, research institutes, scientific journals, companies, farmer, scientists, plant nutrition
Select Language:
 
 
 
 
Main Menu
Please click the main subject to get the list of sub-categories
 
Services offered
 
 
 
 
  Section: Plant Lab Protocols
 
 
Please share with your friends:  
 
 

Methodology for Enzymes

 
     
 
Phosphoenol Pyruvate Carboxylase
(Orthophosphate: Oxaloacetate Carboxylase Phosphorylative EC 4.1.1.31)
 
This enzyme is involved in the fixation of carbon-dioxide in C4 plants. It has a high affinity to CO2 molecule. The reaction catalyzed by this enzyme is given below.
Phosphoenol Pyruvate + CO2
PEP Case
Oxaloacetic acid
 
 
Sorghum, maize, sugarcane etc. are some of C4 plants.
 
 



Principle
The oxaloacetic acid formed by the action of PEP case is coupled with MDH and followed by NADH oxidation. Since the sources of PEP case contain enough MDH there is no need to add MDH exogenously.
 
 



Materials
Tris HCl Buffer 0.1M (pH 7.8)
Magnesium Chloride (10mM) 203.5mg/50mL water.
Sodium Bicarbonate (10mM) 84mg/100mL water
Phosphoenolpyruvate (5mM) 23.28mg/100mL water
NADH (0.4mM) 5.32mg/10mL water
Enzyme extract - see under MDH
 
 
Procedure
1.  Mix various recipes as shown below:
Sr. No.
Reagents
Test
Blank
 
 
mL
mL
1
Tris-HCl buffer pH 7.8
0.8
1.3
2
Magnesium chloride
0.5
0.5
3
Sodium bicarbonate
0.5
0.5
4
Phosphoenol pyruvate
0.5
0.5
5
Enzyme extract
0.2
0.2
6
NADH
0.5
0
2.  Proceed steps 2 to 4 as in MDH
.
 
Calculation
Calculate as for MDH.
 
 


References
1.  Ting I P and Osmond, C B (1973) Plant Physiol 51 439.
2.  Sadasivam, S and Gowri, G (1981) Photosynthetica 15 453.

 
     
 
 
     




     
 
Copyrights 2012 © Biocyclopedia.com | Disclaimer