Biofuels contain alcohols that are produced by the fermentation of sugars derived from crop waste. This waste contains cellulose and other organic compounds in cell walls. Scientists investigated the production of sugars from crop waste for biofuel production. The scientists discovered that a strain of the fungus Penicillium citrinum, isolated from soil, was a good source of three different extracellular enzymes, M, N and O. These enzymes break down polysaccharides in cell walls. The scientists cultured P. citrinum in a liquid medium containing cell wall material. Samples of the liquid were taken, and the three enzymes were separated from the medium. Each enzyme was placed in a reaction mixture with an appropriate substrate. The activity of each enzyme was determined to give a measurement of the quantity of enzyme produced by P. citrinum. The results are shown in Table 4.1. [Table 4.1] Optimum conditions for each enzyme were used to obtain the results in Table 4.1. The conditions were different for each enzyme. The scientists carried out further research so that a solution containing the three enzymes (enzyme mixture) could be used for the most efficient production of sugars from crop waste. Suggest what the scientists needed to find out in their research.

1. (a)Biofuels contain alcohols that are produced by the fermentation of sugars derived from crop waste. This waste contains cellulose and other organic compounds in cell walls. Scientists investigated the production of sugars from crop waste for biofuel production. The scientists discovered that a strain of the fungus Penicillium citrinum, isolated from soil, was a good source of three different extracellular enzymes, M, N and O. These enzymes break down polysaccharides in cell walls. The scientists cultured P. citrinum in a liquid medium containing cell wall material. Samples of the liquid were taken, and the three enzymes were separated from the medium. Each enzyme was placed in a reaction mixture with an appropriate substrate. The activity of each enzyme was determined to give a measurement of the quantity of enzyme produced by P. citrinum. The results are shown in Table 4.1. [Table 4.1] Optimum conditions for each enzyme were used to obtain the results in Table 4.1. The conditions were different for each enzyme. The scientists carried out further research so that a solution containing the three enzymes (enzyme mixture) could be used for the most efficient production of sugars from crop waste. Suggest what the scientists needed to find out in their research.[4]
2. (b)Students investigated the composition of the cell wall of leaf cells of thale cress, Arabidopsis thaliana. The students began by isolating the cell wall components from the rest of the cell material. The students used enzymes extracted from a fungal pathogen of A. thaliana to hydrolyse the cell wall components to smaller molecules. The students prepared a reaction mixture containing the cell wall components and the enzymes. After 24 hours, they separated and identified the smaller molecules found in the reaction mixture. Four types of molecule were identified: • short chains of β-glucose • β-glucose • peptides • amino acids. Explain the presence of these molecules in the reaction mixture after 24 hours of hydrolysis.[4]