Biosynthetic gene cluster identification and biological activity of lucilactaene from Fusarium sp. RK97-94

Figures & data

Figure 1. Analysis of the lucilactaene biosynthetic gene cluster.

(a) Chemical structure of lucilactaene (1), NG-391 (2), fusarin C, and fusarin A. (b) The Δluc5 strain lost the ability to produce lucilactaene and NG-391. The Δluc1 strain lost the ability to produce lucilactaene (peak 1) and NG-391 (peak 2) and accumulated peak 3. The wild-type, Δluc5, and Δluc1 strains were cultured in FDY for three days and analyzed by UPLC/MS. Lucilactaene, NG-391, and peak 3 were detected by measuring absorbance at 365 nm. (c) Comparison of lucilactaene, fusarin C [7], and NG-391 (partial) [8] biosynthetic gene clusters. Sequencing data from the 40-kb region is available from DDBJ/EMBL/GenBank under Accession No. LC515193. d) Structural determination of the mixture of 3 and 4.

Table 1. ¹H and ¹³C NMR chemical shift comparison of lucilactaene and demethylated analogs in CDCl_3.

	Lucilactaene (1)			Demethyllucilactaene (3)			(8Z)-demethyllucilactaene (4)
Position	C (δC)	H (δH)	Multi (J in Hz)	C (δC)	H (δH)	Multi (J in Hz)	C (δC)	H (δH)	Multi (J in Hz)
1	15.9	1.75	dd (1.4, 7)	16.2	1.77	d (7)	16.2	1.77	d (7)
2	140.6	7.01	qd (0.9, 7)	142.6*^1	7.08	q (7)	142.5*^1	7.08	q (7)
3	130.3			129.9			129.9
4	128.1	6.24	s	127.7*^2	6.22	s	127.6*^2	6.22	s
5	138.0			138.7*^3			138.4*^3
6	142.3	6.64	d (15.1)	142.4*^4	6.63	d (15)	142.3*^4	6.63	d (15)
7	128.3	6.47	dd (11, 15.1)	128.5	6.46	dd (11, 15)	123.7	6.94	dd (11, 15)
8	143.7	6.85	dd (11, 14.7)	143.7	6.83	dd (11, 14)	139.6	6.55	t (10)
9	128.0	6.68	dd (11.5, 14.7)	128.1	6.65	dd (12, 15)	124.0	6.38	t (11)
10	145.6	7.48	d (11.5)	145.7	7.46	d (11)	139.7	7.96	d (12)
11	134.2			134.1			134.7
12	197.0			197.1*^5			196.9*^5
13	56.6	4.36	s	57.4*^6	4.36	s	56.7*^6	4.36	s
14	85.7	4.25	d (0.9)	85.7	4.24	s	85.7	4.28	s
15	94.4			94.8			94.8
17	170.5			171.2*^7			171.1*^7
18	37.5	2.28, 2.43	Multi	37.4*^8	2.27, 2.41	Multi	37.5*^8	2.27, 2.41	Multi
19	68.5	4.03, 4.13	Multi	68.5*^9	4.01, 4.11	Multi	68.6*^9	4.01, 4.11	Multi
20	167.5			170.4			170.4
21	52.0	3.75	s	-			-
22	14.3	1.72	d (1.4)	14.6*^10	1.79*^11	s	14.3*^10	1.72*^11	s
23	11.6	1.96	s	11.5	1.94	s	11.3	1.95	s

*1-*11: The assignments may be interchanged.

Table 2. Growth inhibitory activity of lucilactaene, NG-391, and the mixture of 3 and 4 (IC₅₀ values, μg/mL).

	Lucilactaene (1)	NG-391 (2)	3 + 4	Positive controls
Cancer cells
HeLa	10	5.9	>30	0.0036^*1
HL60	7.0	6.6	>30	0.00085^*1
Bacteria
Staphylococcus aureus	>30	>30	>30	0.58^*2
Escherichia coli	>30	>30	>30	0.55^*2
Fungi
Aspergillus fumigatus	>30	>30	>30	<0.0092^*3
Pyricularia oryzae	>30	>30	>30	0.011^*3
Candida albicans	>30	>30	>30	<0.0092^*3
Malaria parasite
Plasmodium falciparum	0.063	0.75	17	0.0085^*4

*1: taxol; *2: chloramphenicol; *3: amphotericin B; *4: artemisinin.

Niehaus EM, Kleigrewe K, Wiemann P, et al. Genetic manipulation of the Fusarium fujikuroi fusarin gene cluster yields insight into the complex regulation and fusarin biosynthetic pathway. Chem Biol. 2013;20:1055–1066.

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Data availability statement

The data described in this article are openly available in the Open Science Framework at DOI:10.17605/OSF.IO/TPA6U.