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Identification of novel tylosin analogues generated by a wblA disruption mutant of Streptomyces ansochromogenes.

Lu C, Liao G, Zhang J, Tan H - Microb. Cell Fact. (2015)

Bottom Line: One wblA homologue was found in Streptomyces ansochromogenes 7100 by using the Basic Local Alignment Search Tool.Two novel tylosin analogues (compound 1 and 2) were generated by ΔwblA.Bioassays showed that compound 1 and 2 displayed much higher activity than tylosin against Streptococcus pneumoniae, implying that these two compounds might be used to widen the application of tylosin.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China. Lucheng522@aliyun.com.

ABSTRACT

Background: Streptomyces, as the main source of antibiotics, has been intensively exploited for discovering new drug candidates to combat the evolving pathogens. Disruption of wblA, an actinobacteria-specific gene controlling major developmental transition, can cause the alteration of phenotype and morphology in many species of Streptomyces. One wblA homologue was found in Streptomyces ansochromogenes 7100 by using the Basic Local Alignment Search Tool. It is interesting to identify whether novel secondary metabolites could be produced by the wblA disruption mutant as evidenced in other Streptomyces.

Results: The wblA disruption mutant of S. ansochromogenes 7100 (ΔwblA) was constructed by homologous recombination. ΔwblA failed to produce spores and nikkomycin, the major product of S. ansochromogenes 7100 (wild-type strain) during fermentation. Antibacterial activity against Staphylococcus aureus and Bacillus cereus was observed with fermentation broth of ΔwblA but not with that of the wild-type strain. To identify the antibacterial compounds, the two compounds (compound 1 and compound 2) produced by ΔwblA were characterized as 16-membered macrolides by mass spectrometry and nuclear magnetic resonance spectroscopy. The chemical structure of these compounds shows similarity with tylosin, and the bioassays indicated that the two compounds inhibited the growth of a number of gram-positive bacteria. It is intriguing that they displayed much higher activity than tylosin against Streptococcus pneumoniae.

Conclusions: Two novel tylosin analogues (compound 1 and 2) were generated by ΔwblA. Bioassays showed that compound 1 and 2 displayed much higher activity than tylosin against Streptococcus pneumoniae, implying that these two compounds might be used to widen the application of tylosin.

No MeSH data available.


Related in: MedlinePlus

Identification of compound 1 and 2 produced by ΔwblA. HPLC chromatograms (a) and the UV absorption spectra of compound 1 and 2 (b). (I): purified compound 1, (II): purified compound 2, (III): fermentation broth from ΔwblA
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Fig4: Identification of compound 1 and 2 produced by ΔwblA. HPLC chromatograms (a) and the UV absorption spectra of compound 1 and 2 (b). (I): purified compound 1, (II): purified compound 2, (III): fermentation broth from ΔwblA

Mentions: Based on the fact that nikkomycin production was abolished in ΔwblA, it is noteworthy to identify whether new products could be produced by ΔwblA. The culture filtrates from the different time-course experiments were subjected to bioassays against representative gram-positive bacteria and gram-negative bacteria (Additional file 1: Table S1). The culture filtrate collected from ΔwblA after incubation for 96 h showed clear inhibition zones against both Staphylococcus aureus and Bacillus cereus, whereas no inhibition zone was found in the culture filtrate from WT (Fig. 3a, b). Chloroform extracts from these cultures were further analyzed by HPLC (Fig. 3c), and distinct peaks appeared at 17 min (compound 1) and 18 min (compound 2) in the extract of ΔwblA (Fig. 4a). Both compounds gave rise to distinctive absorption at wavelength 286 nm on the ultra-violet (UV) spectra (Fig. 4b), indicating that they might be new products generated by ΔwblA since these two compounds were not found in WT under the same conditions.Fig. 3


Identification of novel tylosin analogues generated by a wblA disruption mutant of Streptomyces ansochromogenes.

Lu C, Liao G, Zhang J, Tan H - Microb. Cell Fact. (2015)

Identification of compound 1 and 2 produced by ΔwblA. HPLC chromatograms (a) and the UV absorption spectra of compound 1 and 2 (b). (I): purified compound 1, (II): purified compound 2, (III): fermentation broth from ΔwblA
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4630966&req=5

Fig4: Identification of compound 1 and 2 produced by ΔwblA. HPLC chromatograms (a) and the UV absorption spectra of compound 1 and 2 (b). (I): purified compound 1, (II): purified compound 2, (III): fermentation broth from ΔwblA
Mentions: Based on the fact that nikkomycin production was abolished in ΔwblA, it is noteworthy to identify whether new products could be produced by ΔwblA. The culture filtrates from the different time-course experiments were subjected to bioassays against representative gram-positive bacteria and gram-negative bacteria (Additional file 1: Table S1). The culture filtrate collected from ΔwblA after incubation for 96 h showed clear inhibition zones against both Staphylococcus aureus and Bacillus cereus, whereas no inhibition zone was found in the culture filtrate from WT (Fig. 3a, b). Chloroform extracts from these cultures were further analyzed by HPLC (Fig. 3c), and distinct peaks appeared at 17 min (compound 1) and 18 min (compound 2) in the extract of ΔwblA (Fig. 4a). Both compounds gave rise to distinctive absorption at wavelength 286 nm on the ultra-violet (UV) spectra (Fig. 4b), indicating that they might be new products generated by ΔwblA since these two compounds were not found in WT under the same conditions.Fig. 3

Bottom Line: One wblA homologue was found in Streptomyces ansochromogenes 7100 by using the Basic Local Alignment Search Tool.Two novel tylosin analogues (compound 1 and 2) were generated by ΔwblA.Bioassays showed that compound 1 and 2 displayed much higher activity than tylosin against Streptococcus pneumoniae, implying that these two compounds might be used to widen the application of tylosin.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China. Lucheng522@aliyun.com.

ABSTRACT

Background: Streptomyces, as the main source of antibiotics, has been intensively exploited for discovering new drug candidates to combat the evolving pathogens. Disruption of wblA, an actinobacteria-specific gene controlling major developmental transition, can cause the alteration of phenotype and morphology in many species of Streptomyces. One wblA homologue was found in Streptomyces ansochromogenes 7100 by using the Basic Local Alignment Search Tool. It is interesting to identify whether novel secondary metabolites could be produced by the wblA disruption mutant as evidenced in other Streptomyces.

Results: The wblA disruption mutant of S. ansochromogenes 7100 (ΔwblA) was constructed by homologous recombination. ΔwblA failed to produce spores and nikkomycin, the major product of S. ansochromogenes 7100 (wild-type strain) during fermentation. Antibacterial activity against Staphylococcus aureus and Bacillus cereus was observed with fermentation broth of ΔwblA but not with that of the wild-type strain. To identify the antibacterial compounds, the two compounds (compound 1 and compound 2) produced by ΔwblA were characterized as 16-membered macrolides by mass spectrometry and nuclear magnetic resonance spectroscopy. The chemical structure of these compounds shows similarity with tylosin, and the bioassays indicated that the two compounds inhibited the growth of a number of gram-positive bacteria. It is intriguing that they displayed much higher activity than tylosin against Streptococcus pneumoniae.

Conclusions: Two novel tylosin analogues (compound 1 and 2) were generated by ΔwblA. Bioassays showed that compound 1 and 2 displayed much higher activity than tylosin against Streptococcus pneumoniae, implying that these two compounds might be used to widen the application of tylosin.

No MeSH data available.


Related in: MedlinePlus