Antibacterial and Antifungal Activities of Different Parts of Tagetes patula.: Preparation of Patuletin Derivatives

Figures & data

Scheme 1 Bioassay-guided isolation of antimicrobial principle (5) from involucres of T. patula..

Table 1 Antibacterial activity of different extracts of aerial parts, roots, and seeds of Tagetes patula. plant.

	Zone of inhibition (mm)
	JWP	JWM	JWM-70	JRP	JRM	JRM-70	JSP	JSM	JSM-70
Gram-positive bacteria
Bacillus cereus.	9	—	—	7	9	—	—	—	—
Bacillus subtilis.	8	8	7	10	9	7	9	11	8
Listeria monocytogenes.	ND	—	8	ND	9	ND	—	8	ND
Micrococcus luteus.	7	—	—	7	10	—	—	—	—
Micrococcus lysodeikticus.	7	—	—	7	7	—	—	—	—
Mycobacterium fortuitum. JS	—	—	—	—	—	—	12	—	8
Staphylococcus aureus.	9	—	—	8	7	7	9	8	—
Staphylococcus aureus. AB 188	7	—	—	—	—	—	8	8	7
Staphylococcus saprophyticus.	9	—	8	7	—	7	—	—	—
Streptococcus faecalis.	7	—	—	7	—	—	8	8	8
Streptococcus pneumoniae.	—	ND	ND	7	10	7	—	—	—
Streptococcus pyogenes.	7	ND	ND	7	7	—	—	9	—
Gram-negative bacteria
Escherichia coli.	7	7	7	8	7	8	9	8	8
Klebsiella pneumoniae.	7	10	8	8	—	7	—	—	—
Proteus mirabilis.	7	7	—	8	—	—	—	7	11
Proteus vulgaris.	10	7	8	8	—	7	—	—	—
Pseudomonas aeruginosa. PAO 286	—	7	10	10	—	7	—	—	—
Salmonella paratyphi. A	8	7	—	7	—	—	—	—	—
Salmonella paratyphi. B	7	—	—	7	—	7	—	10	8
Salmonella typhi.	9	7	7	7	7	7	9	8	—
Shigella flexneri.	7	—	—	—	—	—	11	10	20

JW, aerial parts (stems and leaves); JR, roots; JS, Seeds; JWP, petroleum ether (P.E.) extract of JW; JWM, methanol extract of JW; JWM-70, 70% methanol-water extract of JW; JRP, P.E. extract of JR; JRM, Methanol extract of JR; JRM-70, 70% methanol-water extract of JR; JSP, P.E. extract of JS; JSM, methanol extract of JS; JSM-70, 70% methanol-water extract of JS.

–, Inactive; ND, not done.

Note.: All the samples were found to be inactive against Bacillus stearothermophilus. NCTC 10003, Corynebacterium xerosis, Mycobacterium smegmatis., and Staphylococcus epidermidis..

Table 2 Antibacterial activity of different extracts, fractions, pure compounds patuletin (3) and patulitrin (4) from flowers of Tagetes patula., and the derivatives 3a, 3b.

	Zone of inhibition (mm)
	JFP	JFM	JFM-EA	JFM-EA- -13(3)	JFM-EA-14-15c	JFM-Bu (4)	Pat-Bz (3a)	PC (3b)	Benzoic acid (BA)	Cinnamic acid (CA)
Gram-positive bacteria
Bacillus cereus.	7	10	11	13	25	—	—	9	14	12
Bacillus stearothermophilus. NCTC 10003	ND	9	—	—	—	—	ND	ND	—	—
Bacillus subtilis.	8	9	23	12	20	—	—	7	12	12
Bacillus thuringiensis.	—	—	12	8	25	—	—	7	—	8
Corynebacterium hofmanni.	—	—	14	8	23	7	9	10	12	8
Corynebacterium xerosis.	—	9	—	8	7	—	9	10	12	8
Micrococcus luteus.	7	12	22	12	20	9	—	9	11	12
Micrococcus lysodeikticus.	ND	8	—	—	—	ND	ND	ND	—	—
Mycobacterium fortuitum.	—	8	ND	7	ND	ND	ND	ND	—	—
Staphylococcus aureus.	7	—	15	8	23	—	—	7	15	8
Staphylococcus aureus. AB 188	—	10	14	10	21	—	—	7	17	—
Staphylococcus epidermidis.	—	8	19	10	14	—	—	7	15	12
Staphylococcus saprophyticus.	8	10	16	10	25	11	—	7	14	10
Streptococcus faecalis.	—	8	16	8	25	16	—	7	9	8
Streptococcus pneumoniae.	—	—	8	10	22	—	—	7	—	8
Streptococcus pyogenes.	—	20	8	9	24	12	—	7	14	8

	JFP	JFM	JFM-EA	JFMEA-9	JFMEA-13 (3)	JFMEA-14-15c	JFM-Bu (4)	Pat-Bz (3a)	PC (3b)	Benzoic acid (BA)	Cinnamic acid (CA)
Gram-negative bacteria
Enterobacter.	—	—	8	—	7	—	—	—	—	11	8
Escherichia coli.	—	—	—	—	—	—	—	—	9	14	8
Klebsiella pneumoniae.	9	7	—	—	7	—	7	—	—	9	—
Proteus mirabilis.	7	—	—	—	—	—	—	—	9	10	—
Proteus vulgaris.	11	9	—	—	—	—	7	—	8	12	—
Pseudomonas aeruginosa. PAO 286	10	7	—	—	—	—	—	ND	7	—	—
Salmonella paratyphi. A	—	—	7	—	8	—	—	—	12	13	—
Salmonella paratyphi. B	—	—	—	—	7	—	—	—	8	12	—
Salmonella typhi.	—	—	—	—	8	—	—	—	9	10	8
Shigella boydii.	—	—	—	—	—	—	—			11	8
Shigella dysenteriae.	—	—	8	9	—	—	—	—	—	15	8
Shigella flexneri.	—	—	14	—	—	—	—	—	—	10	8

JF, Flowers; JFP, Petroleum ether (P.E.) extract of JF; JFM, Methanol extract of JF; JFM-EA, Ethyl acetate phase of JFM; JFM-EA-9, VLC fraction no. 9 of JFM-EA; JFM-EA-13, Patuletin (3); JFM-EA-14-15c, VLC fraction of JFM-EA; JFM-Bu, Patulitrin (4); Pat-Bz 3, 3′, 4′, 7-tetrabenzoylpatuletin (3a); PC, 3, 3′, 4′, 7-tetracinnamoylpatuletin patuletin (3b).

–, Inactive; ND, not done.

Note.: Listeria monocytogenes. was resistant to the tested samples JFM, JFM-EA, JFM-EA-13, JFM-EA-14-15c, JFM-Bu, and benzoic and cinnamic acids, Mycobacterim smegmatis. was found to be resistant to JFP, JFM, JFM-EA-13, and benzoic and cinnamic acids.

Table 3 Antibacterial activity of involucre, seedlings, and pure compound (5) of Tagetes patula..

	Zone of inhibition (mm)
	JSCP	JSCPMI (5)	JSCEA	JSCEM	JSdEA I	JSdEA II	JSdEA III	JSdM I	JSdM II	JSdM III
Gram-positive bacteria
Bacillus cereus.	8	–	–	–	8	8	8	8	8	8
Bacillus stearothermophillus. NCTC 10003	7	15	–	–	–	–	–	–	–	–
Bacillus subtilis.	7	7	11	8	8	8	–	8	9	8
Corynebacterium xerosis.	–	–	–	–	–	–	–	7	7	8
Micrococcus luteus.	9	9	–	–	10	9	9	9	10	8
Micrococcus lysodeikticus.	–	–	–	–	–	–	8	–	–	–
Mycobacterium fortuitum.	11	–	9	8	–	–	–	–	–	–
Mycobacterium smegmetis.	11	–	9	–	–	–		–	–	–
Staphylococcus aureus.	9	8	8	7	8	9	8	7	8	8
Staphylococcus aureus. AB 188	10	7	8	8	10	–	7	–	7	9
Staphylococcus epidermidis.	10	–	8	–	7	7	–	7	8	–
Staphylococcus saprophyticus.	–	–	–	8	–	7	7	7	7	8
Streptococcus faecalis.	10	9	–	7	9	8	9	8	8	7
Streptococcus pyogenes.	–	–	8	8	8	9	–	7	7	–
Gram-negative bacteria
Escherichia coli.	9	–	–	7	7	7	7	7	7	7
Klebsiella pneumoniae.	–	–	–	–	8	8	7	7	7	7
Proteus mirabilis.	7	7	–	7	8	8	8	8	9	8
Proteus vulgaris.	–	–	–	–	7	7	7	–	7	8
Pseudomonas aeruginosa. PAO 286	–	–	–	–	7	7	7	7	–	–
Salmonella paratyphi. A	8	8	8	–	7	7	10	9	8	7
Salmonella paratyphi. B	8	8	10	9	7	7	8	9	11	9
Salmonella typhi.	8	–	–	9	–	–	–	8	8	9
Shigella boydii.	–	–	–	–	10	–	–	–	–	–
Shigella flexneri.	11	–	8	8	8	8	8	9	9	9

JSC, Dried involucre (without achenes); JSd, Seedlings; JSCP, P.E. extract of JSC; JSCPMI, tetracosanol (5); JSCEA, ethyl acetate (EtOAc) extract of JSC; JSCEM, EtOAc-methanol extract of JSC; JSdEA I, EtOAc extract of JSd (height 11 cm); JSdEA II, EtOAc extract of JSd (height 13 cm); JSdEA III, EtOAc extract of JSd (height 21 cm); JSdM I, methanol extract of JSd (height 11 cm); JSdM II, methanol extract of JSd (height 13 cm); JSdM III, methanol extract of JSd (height 21 cm).

–, inactive.

Note.: All the samples were inactive against Bacillus thuringiensis, Listeria monocytogenes., Corynebacterium hofmanni, Streptococcus pneumoniae, Enterobacter., and Shigella dysenteriae..

Table 4 The MIC (μ g/disk) of fractions and pure compounds.

	Zone of inhibition (mm)
	JFM-EA	JFMEA-13 Patuletin (3)	JFMEA-14a-15c	3,3′,4′,7-Tetrabenzoylpatuletin (3a)	3,3′,4′,7-Tetracinnamoylpatuletin (3b)
Gram-positive bacteria
Bacillus cereus.	–	> 100	> 100	–	> 100
Bacillus subtilis.	> 100	> 100	> 100	–	> 100
Bacillus thuringiensis.	–	> 100	> 100	–	> 100
Corynebacterium hofmanni.	–	12.5	> 100	100	50
Corynebacterium xerosis.	–	12.5	–	100	50
Micrococcus luteus.	> 100	12.5	> 100	–	> 100
Staphylococcus aureus.	> 100	12.5	> 100	–	> 100
Staphylococcus aureus. AB 188	–	> 100	> 100	–	> 100
Staphylococcus epidermidis.	–	50	–	–	> 100
Staphylococcus faecalis.	> 100	12.5	> 100	–	> 100
Staphylococcus pneumoniae.	–	> 100	> 100	–	> 100
Staphylococcus pyogenes.	–	12.5	> 100	–	> 100
Staphylococcus saprophyticus.	> 100	12.5	> 100	–	> 100
Gram-negative bacteria
Escherichia coli.	–	–	–	–	> 100
Enterobacter.	–	100	–	–	> 100
Pseudomonas aeruginosa.	–	–	–	–	> 100
Proteus mirabilis.	–	–	–	–	> 100
Proteus vulgaris.	–	–	–	–	> 100
Salmonella paratyphi. A	–	100	–	–	–
Salmonella paratyphi. B	–	100	–	–	> 100
Salmonella typhi.	–	100	–	–	> 100

JFM-EA, ethyl acetate phase of JFM; JFM-EA-13, patuletin (3); JFM-EA-14-15c, VLC fraction of JFM-EA.

–, inactive.

Table 5 Antifungal activity of different extracts, fractions, and pure compound from Tagetes patula. plant.

	Zone of inhibition (mm)
Fungi	JFM-EA	JFM-EA-13 (3)	JFMEA- 14-15c	Cinnamic acid	JWP	JRP	JSdEAI	JSdEAII	JSdMI	JSdMII	JSdM III
Aspergillus flavus.	10	–	10	–	–	–	10	–	–	–	–
Aspergillus niger.	9	–	9	–	–	–	–	–	–	–	–
Candida albicans.	8	7	9	10	10	10	16	9	9	12	16
Fusarium. sp.	10	–	13	–	–	–	–	–	–	–	–
Helementhosporium.	8	–	8	–	–	–	–	–	–	–	–
Penicillium. sp.	10	–	8	8	–	–	–	–	–	–	–
Rhizopus. sp.	10	–	17	–	–	–	–	–	–	–	–
Saccharomyces cerevisiae.	10	7	11	13	–	–	–	–	–	–	–
Trichophyton mentagrophytes.	24	–	10	–	–	–	–	–	–	–	–
Trichophyton rubrum.	–	–	14	–	–	–	–	–	–	–	–
Trichophyton tonsurans.	–	–	19	–	–	–	–	–	–	–	–

JFM, methanol extract of flower; JFM-EA, ethyl acetate (EtOAc) phase of JFM; JFM-EA-13, patuletin (3); JFM-EA-14-15c, VLC fraction of JFM-EA; JWP, petroleum ether (P.E.) extract of aerial parts; JRP, P.E. extract of roots; JSdEA I, EtOAc extract of seedlings (height 11 cm); JSdEA II, EtOAc extract of seedlings (height 13 cm); JSdM I, methanol extract of seedlings (height 11 cm); JSdM II, methanol extract of seedlings (height 13 cm); JSdM III, methanol extract of seedlings (height 21 cm).

–, inactive.

Note.: Epidermophyton floccossum, Microsporum canis., and Microsporum gypseum. were resistant to the above tested samples.

Scheme 2 Bioassay-guided isolation of antimicrobial principle (3) and (4) from fresh, air-dried uncrushed, orange-red flowers of Tagetes patula. (750 g).

Figure 1 α-Terthienyl.

Figure 2 Methyl protocatechuate.

Figure 3 Patuletin (3) and its tetrabenzoyl (3a) and tetracinnamoyl (3b) derivatives.

Figure 4 Patulitrin.

Figure 5 Tetracosanol.