Compositions and methods for using genetically modified enzymes

A method of producing at least one prenylated product, comprising, a) contacting a first recombinant polypeptide with a substrate and a first prenyl donor, thereby producing a first prenylated product;

• and b) contacting the first prenylated product and a second prenyl donor with a second recombinant polypeptide, thereby producing a second prenylated product,
• wherein the first recombinant polypeptide and/or the second recombinant polypeptide produces a ratio of an amount of the at least one prenylated product to an amount of total prenylated products that is higher than a prenyl transferase comprising the amino acid sequence of SEQ ID NO:
• 1 under the same condition,
• wherein the first recombinant polypeptide and/or the second recombinant polypeptide comprises an amino acid sequence sharing at least 80% sequence identity with the amino acid sequence of SEQ ID NO:
• 1, and
• wherein the first recombinant polypeptide and/or the second recombinant polypeptide comprises:
• (a) two or more amino acid substitutions relative to SEQ ID NO:
• 1 selected from the group consisting of:
• A17T, C25V, Q38G, V49A, V49L, V49S, A53C, A53D, A53E, A53F, A53G, A53H, A53I, A53K, A53L, A53M, A53N, A53P, A53Q, A53R, A53S, A53T, A53V, A53W, A53Y, M106E, A108G, E112D, E112G, K118N, K118Q, K119A, K119D, Y121W, F123A, F123H, F123W, Q161A, Q161C, Q161D, Q161E, Q161F, Q161G, Q161H, Q161I, Q161K, Q161L, Q161M, Q161N, Q161P, Q161R, Q161S, Q161T, Q161V, Q161W, Q161Y, M162A, M162F, D166E, N173D, L174V, S177E, S177W, S177Y, G205L, G205M, C209G, F213M, S214A, S214C, S214D, S214E, S214F, S214G, S214H, S214I, S214K, S214L, S214M, S214N, S214P, S214Q, S214R, S214T, S214V, S214W, S214Y, Y216A, L219F, D227E, R228E, R228Q, C230N, C230S, A232S, V271E, L274V, Y283L, G286E, Y288A, Y288C, Y288D, Y288E, Y288F, Y288G, Y288H, Y288I, Y288K, Y288L, Y288M, Y288N, Y288P, Y288Q, Y288R, Y288S, Y288T, Y288V, Y288W, V294A, V294F, V294N, Q295A, Q295C, Q295D, Q295E, Q295F, Q295G, Q295H, Q295I, Q295K, Q295L, Q295M, Q295N, Q295P, Q295R, Q295S, Q295T, Q295V, Q295W, Q295Y, L298A, L298Q, and L298W;
• or
• (b) two or more sets of amino acid substitutions relative to SEQ ID NO:
• 1 selected from the group consisting of:
• A53T and S214R;
• S177W and Q295A;
• S214R and Q295F;
• Q161S and S214R;
• S177W and S214R;
• Q161S and Q295L;
• Q161S and Q295F;
• V49A and S214R;
• A53T and Q295F;
• Q161S and S177W;
• Q161S, V294A and Q295W;
• A53T, Q161S and Q295W;
• A53T and S177W;
• A53T, Q161S, V294A and Q295W;
• A53T, V294A and Q295A;
• V49A and Q295L;
• A53T, Q161S, V294N and Q295W;
• A53T and Q295A;
• Q161S, V294A and Q295A;
• A53T and Q295W;
• A53T, V294A and Q295W;
• A53T, Q161S and Q295A;
• A53T, Q161S, V294A and Q295A;
• and A53T, Q161S, V294N and Q295A.