Antibacterial activity of phytopathogenic <i>Streptomyces</i> strains against bacteria associated to clinical diseases

Tomaseto, Alex Augusto; Alpiste, Marcel Costa; Nassar, Alessandra Figueiredo de Castro; Destéfano, Suzete Aparecida Lanza

doi:10.1590/1808-1657000142020

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Scientific Article, Plant Pathology • Arq. Inst. Biol. 87 • 2020 • https://doi.org/10.1590/1808-1657000142020 copy

Antibacterial activity of phytopathogenic Streptomyces strains against bacteria associated to clinical diseases^* * This paper is part of the dissertation thesis of the first author.

Atividade antimicrobiana de Streptomyces fitopatogênicas contra bactérias associadas a doenças de importância clínica

Authorship SCIMAGO INSTITUTIONS RANKINGS

ABSTRACT:

The genus Streptomyces is associated with the ability to produce and excrete a variety of bioactive compounds, such as antibiotic, antifungal and antiviral. Biological active polyketide and peptide compounds with applications in medicine, agriculture and biochemical research are synthesized by PKS-I and NRPS genes. The evaluation of the presence of these genes associated with the biosynthesis of secondary metabolites in different phytopathogenic Streptomyces strains were performed using degenerated primers. The positive signal was observed in 58/63 Streptomyces strains for NRPS gene, 43/63 for PKS-I, and for PKS-II all the 63 strains showed positive signal of amplification. These strains also were tested with double layer agar-well technique against bacterial with clinical importance, and it was possible to observe the Streptomyces spp. strains were able to inhibit the growth of 14, 20, 13 and 3 isolates Gram-positive and Gram-negative bacteria, Staphylococcus aureus (ATCC 25923), Bacillus cereus (ATCC 14579), Pseudomonas aeruginosa (ATCC 27853) and Escherichia coli (ATCC 11775) respectively. The Streptomyces sp. strains IBSBF 2019 and IBSBF 2397 showed antibacterial activity against all four bacteria-target tested.

KEYWORDS:
biosynthetic pathway; antimicrobial activity; Streptomyces spp

Gene	Primers	Sequence (5′- 3′)	Fragment (bp)	primers (μM)	dNTPs (mM)	MgCl₂ (mM)	Taq DNA pol	Amplification program
PKS-I	K1	TSAAGTCSAACATCGGBCA	1100	0.4	0.2	1.5	2.0	95°C/5 min; 35X (95°C/30 seg, 58°C/30 seg, 72°C/4 min); 72°C/10 min
PKS-I	M6R	CGCAGGTT SCSGTACCAGTA	1100	0.4	0.2	1.5	2.0
PKS-II	IIPF6	TSGC STGCTTCGAYGCSATC	600-700	0.4	0.2	1.5	2.0	95°C/5 min; 35X (95°C/30 seg, 58°C/30 seg, 72°C/4 min); 72°C/10 min
PKS-II	IIPR6	TGGAANCCGCCGAABCCGCT	600-700	0.4	0.2	1.5	2.0
NRPS	A3	GCSTACSYSATSTACACSTCSGG	700	0.4	0.2	1.5	2.0	95°C/5 min; 35X (95°C/30 seg, 58°C/30 seg, 72°C/4 min); 72°C/10 min
NRPS	A7R	SASGTCVCCSGTSCGGT AS	700	0.4	0.2	1.5	2.0

Genus/Species		Genes			Clinical importance bacteria
	IBSBF	NRPS	PKS-I	PKS-II	S. aureus (ATCC 25923)	B. cereus (ATCC 6051)	P. aeruginosa (ATCC 133388)	E. coli (ATCC 11775)
S. scabiei	1884	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	1886	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.
	1936	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2005	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2006	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.
	2124	^– (-) negative amplification signal.	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2147	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2203	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2204	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2228	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2243	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2248	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2250	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.
	2257	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2282	^– (-) negative amplification signal.	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	–
	2292	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	–	^– (-) negative amplification signal.
	2298	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2315	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.
	2316	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2317	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2359	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	–	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2388	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2403	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2475	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2500	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2501	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2502	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2523	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.
S. europaeiscabiei	1943	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	1944	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2162	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2472	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2473	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2474	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2498	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2499	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2508	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2510	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.
S. caviscabies	2021	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	–	–	–
	2236	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2260	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2368	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2392	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.	–
S. sampsonii	2360	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.
Streptomyces sp.	2019	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;
	2397	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;
	2402	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2430	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.
	2431	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2439	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2449	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.
	2486	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.	–
	2528	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.	–
	2247	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.
	2352	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;
	2390	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2395	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2509	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2520	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.
	2503	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2507	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.
	2229	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.
	2343	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	⁺ (+) positive amplification signal for determinate gene;	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.	^– (-) negative amplification signal.

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Antibacterial activity of phytopathogenic Streptomyces strains against bacteria associated to clinical diseases* * This paper is part of the dissertation thesis of the first author.

Atividade antimicrobiana de Streptomyces fitopatogênicas contra bactérias associadas a doenças de importância clínica

ABSTRACT:

Antibacterial activity of phytopathogenic Streptomyces strains against bacteria associated to clinical diseases^* * This paper is part of the dissertation thesis of the first author.