Patent ID: 8008078
Filing Date: 2011-08-30
Classification: A61P,C12N,C12P

Abstract:
1. An isolated Streptomyces avermitilis cell which produce a cyclohexyl B2:cyclohexyl B1 avermectin ratio of about 0.35:1 or less, comprising a nucleotide sequence that is otherwise the same as the S.avermitilis aveC allele, the S. avermititis AveC gene product-encoding sequence of plasmid pSE 186 (ATCC 209604) or the nucleotide sequence of the aveC ORF of S.avermitilis of SEQ ID NO:1, or a degenerate variant thereof, which nucleotide encodes a functionally equivalent aveC gene product and which nucleotide sequence further comprises mutations encoding one or more amino acid substitutions in the amino acid sequence of SEQ. ID. NO:2 wherein said amino acid substitution is selected from the group consisting of: (a) D48E, A61T, A89T, S138T, A139T, G179S, A198G, P289L; (b) G40S, D48E, L136P, G179S, E238D; (c) D48E, L136P, R163Q, G179S; (d) D48E, L136P, R163Q, G179S, E238D; (e) D48E, L136P, R163Q, G179S, A200G, E238D; (f) D48E, L136P, G179S, E238D; (g) D48E, A61T, L136P, G179S, E238D; (h) D48E, A61T, L136P, G179S; (j) D48E, A61T, L136P, G179S, A198G, P202S, E238D, P289L; (k) D48E, A61T, L136P, S138T, A139F, G179S, E238D, P289L; (l) D48E, L136P, G179S, A198G, E238D, P289L; (m) D48E, A61T, S138T, A139F, G179S, A198G, P289L; (n) D48E, L84P, G111V, S138T, A139T, G179S, A198G, P289L; (o) Y28C, D48E, A61T, A89T, S138T, A139T, G179S, E238D; (p) D48E, A61T, A107T, S108G, L136P, G179S, S192A, E238D, P289L; (q) D48E, L136P, G179S, R250W; (r) D48E, A89T, S138T, A139T, R163Q, G179S; (s) D48E, L136P, G179S, A198G, P289L; (t) D48E, F78L, A89T, L136P, G179S; (u) D48E, A89T, S138T, A139T, G179S, E238D, F278L; (v) D48E, A89T, L136P, R163Q, G179S; (w) D48E, A61T, A89T, G111V, S138T, A139F, G179S, E238D, P289L; (x) D25G, D48E, A89T, L136P, S138T, A139T, V141A, I159T, R163Q, G 179S; (y) D48E, A89T, S90G, L136P, R163Q, G179S, E238D; (z) D48E, A61T, A89T, G111V, S138T, A139T, G179S, E238D, P289L; (ab) D48E, L136P, R163Q, G179S, S231L; (ac) D48E, L136P, S138T, A139F, G179S, V196A, E238D; (ad) D48E, A61T, A89T, F99S, S138T, A139T, G179S, E238D; (ae) G35S, D48E, A89T, S138T, A139T, G179S, P289L; (af) D48E, A61T, A89T, S138T, A139T, G179S, V196A, E238D; (ag) D48E, A89T, G111V, S138T, A139T, G179S, A198G, E238D; (ah) S41G, D48E, A89T, L136P, G179S; (ai) D48E, A89T, L136P, R163Q, G179S, P252S; (aj) D48E, A89T, L136P, G179S, F2345; (ak) D48E, A89T, L136P, R163Q, G179S, E238D; (al) Q36R, D48E, A89T, L136P, G179S, E238D; (am) D48E, A89T, L136P, R163Q, G179S; (an) D48E, A89T, S138T, G179S; (ao) D48E, A89T, L136P, G179S, E238D; (ap) D48E, A89T, L136P, K154E, G179S, E238D; (aq) D48E, A89T, S138T, A139T, K154R, G179S, V196A, P289L; (ar) D48E, A89T, S138T, A139F, G179S, V196A, E238D; (as) D48E, A61T, A89T, L136P, G179S, V196A, A198G, P289L; (at) D48E, A61T, S138T, A139F, G179S, V196A, E238D, P289L; (au) D48E, A89T, L136P, G179S; (av) D48E, A89T, V120A, L136P, G179S; (aw) D48E, A61T, A89T, S138T, A139F, G179S, V196A, A198G, E238D; (ax) D48E, A61T, A89T, G111V, S138T, A139F, G179S, V196A, E238D; (ay) D48E, A61T, A89T, S138T, A139T, G179S, V196A, E238D, P289L; (az) D48E, A61T, A89T, L136P, S138T, A139F, G179S, A198G, E238D; (ba) D48E, A89T, S138T, A139F, G179S, A198G, V220A; (bb) D48E, A61T, A89T, S138T, A139T, G179S, V196A, E238D, R239H, P289L; (bc) D48E, A61T, A89T, L136P, G179S, P289L; (bd) D48E, A89T, S138T, A139T, G179S, V196A, E238D, P289L; (be) D48E, A61T, A89T, S138T, A139F, G179S, V196A, E238D; (da) S41G, D48E, A61T, R71L, A89T, L136M, S138T, A139T, T149S, G179S, V196A, E238D, F278L, P289L; (db) S41G, D48E, A61T, R71L, A89T, L136M, S138T, A139T, T149S, F176C, G179S, V196A, E238D, P289L; (de) D48E, R71L, A89T, L136P, T149S, F176C, G179S, E238D, I280V; (dd) D48E, A61T, R71L, W110L, T149S, G179S, V196A, L206M, E238D, V271A, I280V; (de) D48E, A61T, R71L, A89T, L136M, S138T, A139T, T149S, G179S, V196A, E238D, H279Q, P289L; (df) D48E, A61T, R71L, A89T, L136M, S138T, A139T, T149S, G179S, V196A, E238D, G287E, P289Q; (dg) D48E, A61T, R71L, A89T, L136M, S138T, A139T, T149S, G179S, V196A, E238D, P289L; (dh) D48E, A61T, R71L, A89T, A139T, T149S, F176C, G179S, V196A, E238D, V285G, P289L; (di) Q38R, D48E, A61T, R71L, L87V, A89T, L136M, S138T, A139T, T149S, G179S, V196A, E238D, P289L; (dj) D48E, A61T, L87V, A89T, W110L, S138T, A139T, T149S, G179S, V196A, E238D, P289L; (dk) D48E, A61T, R71L, A89T, L136M, S138T, A139T, T149S, G179S, V196A, E238D, P289L; (dl) D48E, A89T, L136P, K154E, G179S, S231L, E238D; (ea) D48E, A61T, R71L, A89T, L136P, T149S, F176C, G179S, V196A, E238D, I280V; (eb) D48E, R71L, A89T, L136P, T149S, F176C, G179S, E238D, I280V; (ec) Q36P, D48E, A61T, R71L, A89T, L136P, T149S, F176C, G179S, V196A, E238D, I280V; (ed) D48E, A61T, R71L, A89T, A139T, T149S, F176C, G179S, V196A, E238D, I280V; (ee) V2M, D48E, A61T, R71L, A89T, L136P, T149S, F176C, G179S, V196A, E238D, I280V; (ef) D48E, A61T, R71L, A89T, L136P, T149S, F176C, G179S, V196A, E238D, I280V, A302T; (eg) D48E, R71L, A89T, L136P, T149S, F176C, G179S, E238D, P289L; (eh) D48E, R71L, A89T, L136P, T149S, F176C, G179S, E238D, A302T; (ei) D48E, R71L, A89T, L136P, T149S, F176C, G179S, V196A, E238D, I280V; (ej) D48E, A61T, R71L, A89T, L136P, T149S, F176C, G179S, V196A, E238D; (ek) V2M, D48E, R71L, A89T, L136P, T149S, F176C, G179S, V196A, E238D, I280V; (el) D48E, A61T, R71L, A89T, L136P, T149S, R162H, F176C, G179S, V196A, E238D, I280V; (em) D48E, R71L, A89T, V120A, L136P, T149S, K154E, G179S, S231L, E238D; (en) D48E, R71L, A89T, V120A, L136P, T149S, F176C, G179S, S231L, E238D, I280V; (eo) D48E, A61T, R71L, L87V, A89T, S90N, A139T, T149S, F176C, G179S, V196A, E238D, V285G, P289L; (ep) D48E, A61T, R71L, L87V, A89T, A139T, T149S, F176C, G179S, V196A, E238D, V285G, P289L; (eq) D48E, R71L, A89T, L136P, K154E, G179S, S231L, E238D; (er) D48E, R71L, A89T, V120A, L136P, K154E, F176C, G179S, S231L, E238D; and (es) D48E, R71L, A89T, V120A, L136P, T149S, K154E, F176C, G179S, S231L, E238D.