Saccharification of Cellulose Using Acid and Cellulase Enzymes to Produce Cellulosic Ethanol, a Sustainable Fuel

Overview

Objectives/Goals The purpose of my investigation is to develop a method to convert cellulose to glucose utilizing chemical and enzymatic processes based on literature and experimental means. I will work through this process by iterating in successive experiments. Secondly, I want to confirm that the saccharide produced is glucose and fermentable by yeast. Methods/Materials Exp #1: Add H2O to the 1st jar. Add 6 mL conc. H2SO4 and H2O to the 2nd. Add 52 ml conc. H2SO4 to the 3rd. Hold temp. of 80-100 C for >12 hours. Record glucose levels. Adjust pH to 4.8 and temp to 50C . Add 0.7 g of cellulase to first two jars. Hold temp for 48 hours then record glucose. Adjust temp. to 25C and add yeast. Hold for 48 hours. Record Glucose levels. Exp #2: Add H2O to the 1st set of 3 jars. Add 5 mL conc. H2SO4 and H2O to the 2nd set . Add 14 ml conc. H2SO4 to a 3rd set . Repeat for hydrolysis in Exp #1 but use 1.4g of cellulase in all jars. Exp #3: Use paper and 5mL acid steps from Exp#2 for 2 Jars. Place jars in pressure cooker at 15 PSI for 45 minutes. Place in microwave. Record glucose levels. Adjust pH to 4.8 and temp to 50C . Add 2.8 g and 5.6g of cellulase to each jar. Hold temp for 24 hours then record glucose. Adjust temp. to 25C and add yeast. Hold for 48 hours. Record Glucose levels. Exp. #4: Use 5mL acid steps from Exp. #2 for 3 jars of paper and 3 of newsprint. Take pH and add acid until pH is below 2. Repeat steps for hydrolysis in Exp. #3 but use 7g of cellulase in all jars. Materials: Copy and newsprint paper; Sulfuric acid; Cellulase enzymes; Potassium hydroxide; Sodium bicarbonate Results The highest concentration of acid from experiment #1 produced a high amount of total sugars, 5863mg. Due to side reactions, 35% of the sugar produced was not glucose. Using a moderate amount of acid in experiments 2,3 and 4 produced 100% glucose. The amount of glucose produced is correlated to the amount Cellulase used in a relationship of about 1.1:1. Conclusions/Discussion Enzymes are critical for the efficient hydrolysis of cellulose to produce glucose and thus the sustainable fuel, ethanol. The enzyme easily unzipped the cellulose after I spent weeks trying to break the strong alpha bonds in cellulose with heat, pressure, acid and microwaves. Enzymes are amazing. More work needs to be done in this field to produce enough ethanol from cellulose to help our fuel needs.

Awards (1)

• Honorable Mention

Competition history

• CSEF 2010 Biochemistry/ Molecular Biology · Entry J0410

Resources

• CSEF project summary ↗

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