US 2015/0044735 A1 · Biosynthetic Systems Producing Fungal Indole Alkaloids

Abstract

The biosynthesis of fungal bicyclo[2.2.2]diazaoctane indole alkaloids with a wide spectrum of biological activities have attracted increasing interest. Their intriguing mode of assembly has long been proposed to feature a non-ribosomal peptide synthetase, a presumed intramolecular Diels-Alderase, a variant number of prenyltransferases, and a series of oxidases responsible for the diverse tailoring modifications of their cyclodipeptide-based structural core. Until recently, the details of these biosynthetic pathways have remained largely unknown due to lack of information on the fungal derived biosynthetic gene clusters. Herein, we report a comparative analysis of four natural product metabolic systems of a select group of bicyclo[2.2.2]diazaoctane indole alkaloids including (+)/(−)-notoamide, paraherquamide and malbrancheamide, in which we propose an enzyme for each step in the biosynthetic pathway based on deep annotation and on-going biochemical studies.

Claims12

Dependent claims

1-6. (canceled)

A host cell transformed with one or more polynucleotides selected from the group consisting of:

a polynucleotide encoding SEQ ID NO:
3 or a protein 95% identical to SEQ ID NO:
3 having MalA activity,
a polynucleotide encoding SEQ ID NO:
5 or a protein 95% identical to SEQ ID NO:
5 having MalB activity,
a polynucleotide encoding SEQ ID NO:
7 or a protein 95% identical to SEQ ID NO:
7 having MalC activity,
a polynucleotide encoding SEQ ID NO:
9 or a protein 95% identical to SEQ ID NO:
9 having MalD activity,
a polynucleotide encoding SEQ ID NO:
11 or a protein 95% identical to SEQ ID NO:
11 having MalE activity,
a polynucleotide encoding SEQ ID NO:
13 or a protein 95% identical to SEQ ID NO:
13 having MalF activity,
a polynucleotide encoding SEQ ID NO:
15 or a protein 95% identical to SEQ ID NO:
15 having MalG activity,
a polynucleotide encoding SEQ ID NO:
18 or a protein 95% identical to SEQ ID NO:
18 having NotA activity,
a polynucleotide encoding SEQ ID NO:
20 or a protein 95% identical to SEQ ID NO:
20 having NotB activity,
a polynucleotide encoding SEQ ID NO:
22 or a protein 95% identical to SEQ ID NO:
22 having NotC activity,
a polynucleotide encoding SEQ ID NO:
24 or a protein 95% identical to SEQ ID NO:
24 having NotD activity,
a polynucleotide encoding SEQ ID NO:
26 or a protein 95% identical to SEQ ID NO:
26 having NotE activity,
a polynucleotide encoding SEQ ID NO:
28 or a protein 95% identical to SEQ ID NO:
28 having NotF activity,
a polynucleotide encoding SEQ ID NO:
30 or a protein 95% identical to SEQ ID NO:
30 having NotG activity,
a polynucleotide encoding SEQ ID NO:
32 or a protein 95% identical to SEQ ID NO:
32 having NotH activity,
a polynucleotide encoding SEQ ID NO:
34 or a protein 95% identical to SEQ ID NO:
34 having NotI activity,
a polynucleotide encoding SEQ ID NO:
36 or a protein 95% identical to SEQ ID NO:
36 having NotJ activity,
a polynucleotide encoding SEQ ID NO:
38 or a protein 95% identical to SEQ ID NO:
38 having NotK activity,
a polynucleotide encoding SEQ ID NO:
40 or a protein 95% identical to SEQ ID NO:
40 having NotL activity,
a polynucleotide encoding SEQ ID NO:
42 or a protein 95% identical to SEQ ID NO:
42 having NotM activity,
a polynucleotide encoding SEQ ID NO:
44 or a protein 95% identical to SEQ ID NO:
44 having NotN activity,
a polynucleotide encoding SEQ ID NO:
46 or a protein 95% identical to SEQ ID NO:
46 having NotO activity,
a polynucleotide encoding SEQ ID NO:
48 or a protein 95% identical to SEQ ID NO:
48 having NotP activity,
a polynucleotide encoding SEQ ID NO:
50 or a protein 95% identical to SEQ ID NO:
50 having NotQ activity,
a polynucleotide encoding SEQ ID NO:
52 or a protein 95% identical to SEQ ID NO:
52 having NotR activity,
a polynucleotide encoding SEQ ID NO:
55 or a protein 95% identical to SEQ ID NO:
55 having phqA activity,
a polynucleotide encoding SEQ ID NO:
57 or a protein 95% identical to SEQ ID NO:
57 having phqB activity,
a polynucleotide encoding SEQ ID NO:
59 or a protein 95% identical to SEQ ID NO:
59 having phqC activity,
a polynucleotide encoding SEQ ID NO:
61 or a protein 95% identical to SEQ ID NO:
61 having phqD activity,
a polynucleotide encoding SEQ ID NO:
63 or a protein 95% identical to SEQ ID NO:
63 having phqE activity,
a polynucleotide encoding SEQ ID NO:
65 or a protein 95% identical to SEQ ID NO:
65 having phqF activity,
a polynucleotide encoding SEQ ID NO:
67 or a protein 95% identical to SEQ ID NO:
67 having phqG activity,
a polynucleotide encoding SEQ ID NO:
69 or a protein 95% identical to SEQ ID NO:
69 having phD2 activity,
a polynucleotide encoding SEQ ID NO:
71 or a protein 95% identical to SEQ ID NO:
71 having phqI activity,
a polynucleotide encoding SEQ ID NO:
73 or a protein 95% identical to SEQ ID NO:
73 having phqJ activity,
a polynucleotide encoding SEQ ID NO:
75 or a protein 95% identical to SEQ ID NO:
75 having phqK activity,
a polynucleotide encoding SEQ ID NO:
77 or a protein 95% identical to SEQ ID NO:
77 having phqL activity,
a polynucleotide encoding SEQ ID NO:
79 or a protein 95% identical to SEQ ID NO:
79 having phqM activity,
a polynucleotide encoding SEQ ID NO:
81 or a protein 95% identical to SEQ ID NO:
81 having phqN activity, and
a polynucleotide encoding SEQ ID NO:
83 or a protein 95% identical to SEQ ID NO:
83 having phqO activity.

117 more elements

A protein in the prenylated indole alkaloid pathway, wherein the protein is

(a) An MalA protein having the amino acid sequence set out in SEQ ID NO:
3 or a protein 95% identical to SEQ ID NO:
3 having MalA activity;
(b) a MalB protein having the amino acid sequence set out in SEQ ID NO:
5 or a protein 95% identical to SEQ ID NO:
5 having EtuA2 activity;
(c) a MalC protein having the amino acid sequence set out in SEQ ID NO:
7 or a protein 95% identical to SEQ ID NO:
7 having MalC activity;
(d) a MalD protein having the amino acid sequence set out in SEQ ID NO:
9 or a protein 95% identical to SEQ ID NO:
9 having MalD activity;
(e) a MalE protein having the amino acid sequence set out in SEQ ID NO:
11 or a protein 95% identical to SEQ ID NO:
11 having MalE activity;
(f) a MalF protein having the amino acid sequence set out in SEQ ID NO:
13 or a protein 95% identical to SEQ ID NO:
13 having MalF activity;
(g) a MalG protein having the amino acid sequence set out in SEQ ID NO:
15 or a protein 95% identical to SEQ ID NO:
15 having MalG activity;
(h) a NoA protein having the amino acid sequence set out in SEQ ID NO:
18 or a protein 95% identical to SEQ ID NO:
18 having NotA activity;
(i) a NotB protein having the amino acid sequence set out in SEQ ID NO:
20 or a protein 95% identical to SEQ ID NO:
20 having NotB activity;
(j) a NotC protein having the amino acid sequence set out in SEQ ID NO:
22 or a protein 95% identical to SEQ ID NO:
22 having NotC activity;
(k) a NotD protein having the amino acid sequence set out in SEQ ID NO:
24 or a protein 95% identical to SEQ ID NO:
24 having NotD activity;
(l) a NotE protein having the amino acid sequence set out in SEQ ID NO:
26 or a protein 95% identical to SEQ ID NO:
26 having NotE activity;
(m) a NotF protein having the amino acid sequence set out in SEQ ID NO:
28 or a protein 95% identical to SEQ ID NO:
28 having NotF activity;
(n) a NotG protein having the amino acid sequence set out in SEQ ID NO:
30 or a protein 95% identical to SEQ ID NO:
30 having NotG activity;
(o) a NotH protein having the amino acid sequence set out in SEQ ID NO:
32 or a protein 95% identical to SEQ ID NO:
32 having NotH activity;
(p) a NotI protein having the amino acid sequence set out in SEQ ID NO:
34 or a protein 95% identical to SEQ ID NO:
34 having NotI activity;
(q) a NotJ protein having the amino acid sequence set out in SEQ ID NO:
36 or a protein 95% identical to SEQ ID NO:
36 having NotJ activity;
(r) a NotK protein having the amino acid sequence set out in SEQ ID NO:
38 or a protein 95% identical to SEQ ID NO:
38 having NotK activity;
(s) a NotL protein having the amino acid sequence set out in SEQ ID NO:
40 or a protein 95% identical to SEQ ID NO:
40 having NotL activity;
(t) a NotM protein having the amino acid sequence set out in SEQ ID NO:
42 or a protein 95% identical to SEQ ID NO:
42 having NotM activity;
(u) a NotN protein having the amino acid sequence set out in SEQ ID NO:
44 or a protein 95% identical to SEQ ID NO:
44 having NotN activity;
(v) a NotO protein having the amino acid sequence set out in SEQ ID NO:
46 or a protein 95% identical to SEQ ID NO:
46 having EtuT activity;
(w) a NotP protein having the amino acid sequence set out in SEQ ID NO:
48 or a protein 95% identical to SEQ ID NO:
48 having NotP activity;
(x) a NotQ protein having the amino acid sequence set out in SEQ ID NO:
50 or a protein 95% identical to SEQ ID NO:
50 having NotQ activity;
(y) a NotR protein having the amino acid sequence set out in SEQ ID NO:
52 or a protein 95% identical to SEQ ID NO:
52 having NotR activity;
(z) a phqA protein having the amino acid sequence set out in SEQ ID NO:
55 or a protein 95% identical to SEQ ID NO:
24 having phqA activity;
(aa) a phqB protein having the amino acid sequence set out in SEQ ID NO:
57 or a protein 95% identical to SEQ ID NO:
57 having phqB activity;
(ab) a phqC protein having the amino acid sequence set out in SEQ ID NO:
59 or a protein 95% identical to SEQ ID NO:
59 having phqC activity;
(ac) a phqD protein having the amino acid sequence set out in SEQ ID NO:
61 or a protein 95% identical to SEQ ID NO:
61 having phqD activity;
(ad) a phqE protein having the amino acid sequence set out in SEQ ID NO:
63 or a protein 95% identical to SEQ ID NO:
63 having phqE activity;
(ae) a phqF protein having the amino acid sequence set out in SEQ ID NO:
65 or a protein 95% identical to SEQ ID NO:
65 having phqF activity;
(af) a phqG protein having the amino acid sequence set out in SEQ ID NO:
67 or a protein 95% identical to SEQ ID NO:
67 having phqH activity;
(ag) a phqH protein having the amino acid sequence set out in SEQ ID NO:
69 or a protein 95% identical to SEQ ID NO:
69 having phqH activity;
(ah) a phqI protein having the amino acid sequence set out in SEQ ID NO:
71 or a protein 95% identical to SEQ ID NO:
71 having phqI activity;
(ai) a phqJ protein having the amino acid sequence set out in SEQ ID NO:
73 or a protein 95% identical to SEQ ID NO:
73 having phqJ activity;
(aj) a phqK protein having the amino acid sequence set out in SEQ ID NO:
75 or a protein 95% identical to SEQ ID NO:
75 having phqK activity;
(ak) a phqL protein having the amino acid sequence set out in SEQ ID NO:
77 or a protein 95% identical to SEQ ID NO:
77 having phqL activity;
(al) a phqM protein having the amino acid sequence set out in SEQ ID NO:
79 or a protein 95% identical to SEQ ID NO:
79 having phqM activity;
(am) a phqN protein having the amino acid sequence set out in SEQ ID NO:
81 or a protein 95% identical to SEQ ID NO:
81 having phqN activity;
and
(an) a phqO protein having the amino acid sequence set out in SEQ ID NO:
83 or a protein 95% identical to SEQ ID NO:
83 having phqO activity.

118 more elements

A polynucleotide encoding a protein in the prenylated indole alkaloid pathway, wherein the polynucleotide is

(a) a polynucleotide set out in SEQ ID NO:
2 or a polynucleotide 96% homologous thereto
(b) a polynucleotide set out in SEQ ID NO:
4 or a polynucleotide 96% homologous thereto;
(c) a polynucleotide set out in SEQ ID NO:
6 or a polynucleotide 96% homologous thereto;
(d) a polynucleotide set out in SEQ ID NO:
8 or a polynucleotide 96% homologous thereto;
(e) a polynucleotide set out in SEQ ID NO:
10 or a polynucleotide 96% homologous thereto;
(f) a polynucleotide set out in SEQ ID NO:
12 or a polynucleotide 96% homologous thereto;
(g) a polynucleotide set out in SEQ ID NO:
14 or a polynucleotide 96% homologous thereto;
(h) a polynucleotide set out in SEQ ID NO:
17 or a polynucleotide 96% homologous thereto;
(i) a polynucleotide set out in SEQ ID NO:
19 or a polynucleotide 96% homologous thereto;
(j) a polynucleotide set out in SEQ ID NO:
21 or a polynucleotide 96% homologous thereto;
(k) a polynucleotide set out in SEQ ID NO:
23 or a polynucleotide 96% homologous thereto;
(l) a polynucleotide set out in SEQ ID NO:
25 or a polynucleotide 96% homologous thereto;
(m) a polynucleotide set out in SEQ ID NO:
27 or a polynucleotide 96% homologous thereto;
(n) a polynucleotide set out in SEQ ID NO:
29 or a polynucleotide 96% homologous thereto;
(o) a polynucleotide set out in SEQ ID NO:
31 or a polynucleotide 96% homologous thereto;
(p) a polynucleotide set out in SEQ ID NO:33 or a polynucleotide 96% homologous thereto;
(q) a polynucleotide set out in SEQ ID NO:35 or a polynucleotide 96% homologous thereto;
(r) a polynucleotide set out in SEQ ID NO:37 or a polynucleotide 96% homologous thereto;
(s) a polynucleotide set out in SEQ ID NO:
39 or a polynucleotide 96% homologous thereto;
(t) a polynucleotide set out in SEQ ID NO:
41 or a polynucleotide 96% homologous thereto;
(u) a polynucleotide set out in SEQ ID NO:
43 or a polynucleotide 96% homologous thereto;
(v) a polynucleotide set out in SEQ ID NO:
45 or a polynucleotide 96% homologous thereto;
(w) a polynucleotide set out in SEQ ID NO:
47 or a polynucleotide 96% homologous thereto;
(x) a polynucleotide set out in SEQ ID NO:
49 or a polynucleotide 96% homologous thereto;
(y) a polynucleotide set out in SEQ ID NO:
51 or a polynucleotide 96% homologous thereto;
(z) a polynucleotide set out in SEQ ID NO:
54 or a polynucleotide 96% homologous thereto;
(aa) a polynucleotide set out in SEQ ID NO:
56 or a polynucleotide 96% homologous thereto;
(ab) a polynucleotide set out in SEQ ID NO:
58 or a polynucleotide 96% homologous thereto;
(ac) a polynucleotide set out in SEQ ID NO:
60 or a polynucleotide 96% homologous thereto;
(ad) a polynucleotide set out in SEQ ID NO:
62 or a polynucleotide 96% homologous thereto;
(ae) a polynucleotide set out in SEQ ID NO:
64 or a polynucleotide 96% homologous thereto;
(af) a polynucleotide set out in SEQ ID NO:
66 or a polynucleotide 96% homologous thereto;
(ag) a polynucleotide set out in SEQ ID NO:68 or a polynucleotide 96% homologous thereto;
(ah) a polynucleotide set out in SEQ ID NO:
70 or a polynucleotide 96% homologous thereto;
(ai) a polynucleotide set out in SEQ ID NO:
72 or a polynucleotide 96% homologous thereto;
(aj) a polynucleotide set out in SEQ ID NO:
74 or a polynucleotide 96% homologous thereto;
(ak) a polynucleotide set out in SEQ ID NO:
76 or a polynucleotide 96% homologous thereto;
(al) a polynucleotide set out in SEQ ID NO:
78 or a polynucleotide 96% homologous thereto;
(am) a polynucleotide set out in SEQ ID NO:
80 or a polynucleotide 96% homologous thereto;
and
(an) a polynucleotide set out in SEQ ID NO:
82 or a polynucleotide 96% homologous thereto.

74 more elements

Dependencies are inferred from the claim text.

Biosynthetic Systems Producing Fungal Indole Alkaloids

Life of this patent

Biosynthetic Systems Producing Fungal Indole Alkaloids

Life of this patent

Abstract

Claims12