Podcast
Questions and Answers
How does the hydrophobic environment near the haem group of the HRP basic isoenzyme affect its interaction with surfactants?
How does the hydrophobic environment near the haem group of the HRP basic isoenzyme affect its interaction with surfactants?
- It inhibits complex formation with the surfactant
- It has no effect on complex formation with the surfactant
- It favors complex formation with the surfactant, especially at higher protein concentrations (correct)
- It requires the addition of citrate to facilitate the forward transfer process
What is the primary function of the surfactant shell layer in reverse micelles?
What is the primary function of the surfactant shell layer in reverse micelles?
- To increase the citrate concentration and facilitate forward transfer
- To chelate Ca2+ ions and improve protein purification
- To solubilize water and macromolecules within the hydrophilic core
- To protect enzymic proteins from denaturation by the organic phase (correct)
What is a potential biotechnological application of reverse micelles?
What is a potential biotechnological application of reverse micelles?
- Solubilization of hydrophilic proteins, nucleic acids, and other entities (correct)
- Denaturation of enzymic proteins to reduce their catalytic activity
- Chelation of Ca2+ ions to improve protein purification
- Increasing citrate concentration to facilitate forward transfer
What is the effect of back extraction on the activity of proteins solubilized in reverse micelles?
What is the effect of back extraction on the activity of proteins solubilized in reverse micelles?
What is the effect of the organic phase on the catalytic activity of enzymic proteins in reverse micelles?
What is the effect of the organic phase on the catalytic activity of enzymic proteins in reverse micelles?
What is the purpose of using reverse micelles in protein purification?
What is the purpose of using reverse micelles in protein purification?
What was the primary purpose of the first stage of the forward transfer process?
What was the primary purpose of the first stage of the forward transfer process?
What effect did adding citrate to the crude extract have on the purification process?
What effect did adding citrate to the crude extract have on the purification process?
Why was solubilization of contaminant proteins favored over HRP at pH 2.8?
Why was solubilization of contaminant proteins favored over HRP at pH 2.8?
What was the purpose of adding citrate to the crude extract?
What was the purpose of adding citrate to the crude extract?
What conditions were used for the second forward transfer to solubilize HRP?
What conditions were used for the second forward transfer to solubilize HRP?
What was the effect of increasing ionic strength on HRP solubilization at pH 2.8?
What was the effect of increasing ionic strength on HRP solubilization at pH 2.8?
What surfactant was used to produce the reverse-micellar phases in the purification of horseradish peroxidase?
What surfactant was used to produce the reverse-micellar phases in the purification of horseradish peroxidase?
In the purification process, at what specific pH range did HRP solubilization occur?
In the purification process, at what specific pH range did HRP solubilization occur?
What was the final specific activity of the purified horseradish peroxidase (HRP)?
What was the final specific activity of the purified horseradish peroxidase (HRP)?
Which electrophoresis technique showed two overlapping bands corresponding to HRP at 43.8 kDa?
Which electrophoresis technique showed two overlapping bands corresponding to HRP at 43.8 kDa?
Which liquid chromatography technique revealed that most of the specific activity of the purified HRP was due to the basic isoenzyme with pI 8.5?
Which liquid chromatography technique revealed that most of the specific activity of the purified HRP was due to the basic isoenzyme with pI 8.5?
What caused high HRP losses during direct reverse-micellar extraction from the crude extract?
What caused high HRP losses during direct reverse-micellar extraction from the crude extract?
