Questions: Bioprocess for biomass utilization for lactic acid production

Question

Yeast cells are engineered with cellulolytic enzymes that hydrolyze cellulose to glucose. The enzymes are extracted from yeast culture, purified, and then added to a reactor containing cellulose fibers. The reaction mixture is stirred, and the reaction is arrested in 8 hours. The reaction mixture is then filtered to remove unreacted cellulose. The glucose is present in the filtrate along with the enzymes.

1.1 How will you purify glucose from the enzymes?

1.2 How will you determine the hydrolysis yield?

1.3 One batch of cellulose was received contaminated with hemicellulose. You do not know if the enzymes can hydrolyze the hemicellulose. Which experiments will you plan to determine whether to use the batch of contaminated cellulose?

Answer 1

1. So, now you have enzyme and glucose in mixture. you can separate enzymes from the glucose by using different analytical chemistry method. You can use ULTRAFILTRATION, where you can use specific molecular cut off weight to retentate the enzyme. You can use chromatographic techniques to separate the enzymes. As enzymes are mostly proteins so you can use SEC or ion exchange chromatography techniques.
2. Use HPLC method to determine the concentration of glucose.
3. Do the enzyme activity assays.