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Biofuel Lab

Biofuel Lab. Factors that affect cellobiase. Cellobiase activity:. Cellobiose glucose + glucose (substrate) cellobiase (enzyme) (products).

camilla-farren
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Biofuel Lab

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  1. Biofuel Lab Factors that affect cellobiase

  2. Cellobiase activity: Cellobiose glucose + glucose (substrate) cellobiase (enzyme) (products) Unfortunately, we can’t easily detect either the substrate or the products of this reaction.

  3. Cellobiase also works on an artificial substrate: p-Nitrophenyl glucopyranoside Glucose + p-Nitrophenol (substrate) cellobiase (enzyme) (products) • A strong base will denature the enzyme and stop the reaction. • p-Nitrophenol turns yellow in the presence of a base. • More yellow means more p-Nitrophenol. • More p-Nitrophenol means more of the substrate was broken apart.

  4. Part 1: Rate of reaction • Set up 7 cuvettes of stop solution (strong base) – 2 of these are controls • Add enzyme solution to substrate solution and start timing • At intervals of 1, 2, 4, 6 and 8 minutes, transfer 500 uL of the enzyme/substrate mix to a stop solution cuvette • Record absorbance data on each cuvette

  5. Likely hypothesis: As time increases, products of the reaction will increase. Control 1 minute 2 min 4 min 6 min 8 min Control no enzyme no enzyme 0 minutes 8 min Level of yellow can be quantified with a spectrophotometer – more absorbance means more yellow, which means more product

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