Streptomyces have been reported as a remarkable source for bioactive secondary metabolites with complex structural and functional diversity. In this study, 35 isolates of genus Streptomyces were purified from rhizospheric and marine soils collected from previously unexplored habitats and screened for antimicrobial activities. One of these isolates, G1, when tested in vitro, was found highly active against wide range of microbes including Gram-positive, Gram-negative bacteria, and different fungal pathogens. It was identified as mesophilic, alkaliphilic, and moderately halotolerant as it showed optimum growth at temperature 30 °C, pH 8.0 in casein-starch-peptone-yeast extract-malt extract medium supplemented with 5% NaCl. Sequence analysis of the 16S rRNA gene indicated 100% identity of this isolate to Streptomyces fimbriatus. Moreover, maximum antimicrobial activity was achieved in starch nitrate medium supplemented with 1% glycerol as carbon and 0.03% soy meal as nitrogen source. The antimicrobial compounds produced by this isolate were extracted in methanol. Bioassay-guided fractionation through thin layer chromatography of methanolic extract resulted in the separation of a most active fraction with an Rf value of 0.46. This active fraction was characterized by FTIR and LCMS analysis and found similar to streptothricin D like antibiotic with m/z 758.42.

Department of Pharmaceutical Biology and Biotechnology Institute of Pharmaceutical Sciences University of Freiburg Freiburg im Breisgau Germany

Industrial Biotechnology Division National Institute for Biotechnology and Genetic Engineering PO Box 577 Jhang Road Faisalabad Pakistan

See more in PubMed

Ahmad MS, El-Gendy AO, Ahmed RR, Hassan HM, El-Kabbany HM, Merdash AG (2017) Exploring the antimicrobial and antitumor potentials of Streptomyces sp. AGM12–1 isolated from Egyptian soil. Front Microbiology 8:438.  https://doi.org/10.3389/fmicb.2017.00438

Anukool U, Gaze WH, Wellington EM (2004) In situ monitoring of streptothricin production by Streptomyces rochei F20 in soil and rhizosphere. App Environ Microbiol 70(9):5222–5228. https://doi.org/10.1128/AEM.70.9.5222-5228.2004 DOI

Arul Jose P, Satheeja Santhi V, Jebakumar SR (2011) Phylogenetic‐affiliation, antimicrobial potential and PKS gene sequence analysis of moderately halophilic Streptomyces sp. inhabiting an Indian saltpan. J Basic Microbiol 51 (4):348–356.  https://doi.org/10.1002/jobm.201000253

Bérdy J (2012) Thoughts and facts about antibiotics: where we are now and where we are heading. J Antibiot 65(8):385–395. https://doi.org/10.1038/ja.2012.27 DOI

Betancur LA, Naranjo-Gaybor SJ, Vinchira-Villarraga DM, Moreno-Sarmiento NC, Maldonado LA, Suarez-Moreno ZR, Acosta-González A, Padilla-Gonzalez GF, Puyana M, Castellanos L (2017) Marine Actinobacteria as a source of compounds for phytopathogen control: an integrative metabolic-profiling/bioactivity and taxonomical approach. PLoS One 12(2):e0170148. https://doi.org/10.1371/journal.pone.0170148 PubMed DOI PMC

Bhave S, Shanbhag P, Sonawane S, Parab R, Mahajan G (2013) Isolation and characterization of halotolerant Streptomyces radiopugnans from Antarctica soil. Lett App Microbiol 56(5):348–355. https://doi.org/10.1111/lam.12054 DOI

Castro JF, Razmilic V, Gomez-Escribano JP, Andrews B, Asenjo JA, Bibb MJ (2015) Identification and heterologous expression of the chaxamycin biosynthesis gene cluster from Streptomyces leeuwenhoekii. App Environ Microbiol 81 (17):5820–5831.

Challis GL, Hopwood DA (2003) Synergy and contingency as driving forces for the evolution of multiple secondary metabolite production by Streptomyces species. Proceedings of the Nat Acad of Sci 100(suppl 2):14555–14561. https://doi.org/10.1128/AEM.01039-15 DOI

Cumsille A, Undabarrena A, González V, Claverías F, Rojas C, Cámara B (2017) Biodiversity of actinobacteria from the South Pacific and the assessment of Streptomyces chemical diversity with metabolic profiling. Mar Drugs 15(9):286. https://doi.org/10.3390/md15090286 DOI PMC

Donadio S, Monciardini P, Sosio M (2007) Polyketide synthases and nonribosomal peptide synthetases: the emerging view from bacterial genomics. Nat Prod Rep 24(5):1073–1109. https://doi.org/10.1039/b514050c PubMed DOI

Fernández-Moreno MA, Vallin C, Malpartida F (1997) Streptothricin biosynthesis is catalyzed by enzymes related to nonribosomal peptide bond formation. J Bacteriol 179(22):6929–6936. https://doi.org/10.1128/JB.179.22.6929-6936.1997 PubMed DOI PMC

Forar LR, Ali E, Mahmoud S, Bengraa CH, Hacene H (2007) Screening, isolation and characterization of a novel antimicrobial producing actinomycete, strain RAF10. Biotechnol 6 (4): 489–496. https://scialert.net/abstract/?doi=biotech.2007.489.496

Gordon RE, Barnett DA, Handerhan JE, Pang CH-N (1974) Nocardia coeliaca, Nocardia autotrophica, and the nocardin strain. Int J System Evol Microbiol 24(1):54–63. https://doi.org/10.1099/00207713-24-1-54 DOI

Grammel N, Pankevych K, Demydchuk J, Lambrecht K, Saluz HP, Keller U, Krügel H (2002) A β-lysine adenylating enzyme and a β-lysine binding protein involved in poly β-lysine chain assembly in nourseothricin synthesis in Streptomyces noursei. Europ J Biochem 269(1):347–357. https://doi.org/10.1046/j.0014-2956.2001.02657.x PubMed DOI

Horinouchi S, Furuya K, Nishiyama M, Suzuki H, Beppu T (1987) Nucleotide sequence of the streptothricin acetyltransferase gene from Streptomyces lavendulae and its expression in heterologous hosts. J Bacteriol 169(5):1929–1937. https://doi.org/10.1128/jb.169.5.1929-1937.1987 PubMed DOI PMC

Huck TA, Porter N, Bushell ME (1991) Positive selection of antibiotic-producing soil isolates. Microbiol 137(10):2321–2329. https://doi.org/10.1099/00221287-137-10-2321 DOI

Ji Z, Wei S, Zhang J, Wu W, Wang M (2008) Identification of streptothricin class antibiotics in the early stage of antibiotics screening by electrospray ionization mass spectrometry. J Antibiot 61(11):660–667. https://doi.org/10.1038/ja.2008.93 DOI

Jonsbu E, Mcintyre M, Nielsen J (2002) The influence of carbon sources and morphology on nystatin production by Streptomyces noursei. J Biotechnol 95(2):133–144. https://doi.org/10.1016/s0168-1656(02)00003-2 PubMed DOI

Khaliq S, Ghauri MA, Akhtar K (2013) Isolation, identification and optimization of fermentation parameters for improved production of antimicrobial compounds from indigenous Streptomyces isolates. Afr J Microbiol Res 7(18):1874–1887. https://doi.org/10.5897/AJMR2012.2462 DOI

Kim BT, Lee JY, Lee YY, Kim OY, Chu JH, Goo YM (1994) N-Methylstreptothricin DA new streptothricin-group antibiotic from a Streptomyces spp. J Antibiot 47(11):1333–1336. https://doi.org/10.7164/antibiotics.47.1333 DOI

Kumar S, Stecher G, Tamura K (2016) MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol 33 (7):1870–1874.  https://doi.org/10.1093/molbev/msw054

Küster E, Williams S (1964) Selection of media for isolation of Streptomycetes. Nat 202(4935):928–929. https://doi.org/10.1038/202928a0 DOI

Ladurner A, Zehl M, Grienke U, Hofstadler C, Faur N, Pereira FC, Berry D, Dirsch VM, Rollinger JM (2017) Allspice and clove as source of triterpene acids activating the G protein-coupled bile acid receptor TGR5. Front Pharmacol 8:468. https://doi.org/10.3389/fphar.2017.00468 PubMed DOI PMC

Lechevalier MP, Lechevalier H (1970) Chemical composition as a criterion in the classification of aerobic actinomycetes. Int J System Evol Microbiol 20(4):435–443. https://doi.org/10.1099/00207713-20-4-435 DOI

Li J, Guo Z, Huang W, Meng X, Ai G, Tang G, Chen Y (2013) Mining of a streptothricin gene cluster from Streptomyces sp. TP-A0356 genome via heterologous expression. Sci Chi Life Sci 56 (7):619–627.  https://doi.org/10.1007/s11427-013-4504-2

Liu D, Coloe S, Baird R, Pedersen J (2000) Rapid mini-preparation of fungal DNA for PCR. J Clinic Microbiol 38(1):471–471 DOI

Manivasagan P, Venkatesan J, Sivakumar K, Kim S-K (2014) Pharmaceutically active secondary metabolites of marine actinobacteria. Microbiol Res 169(4):262–278. https://doi.org/10.1016/j.micres.2013.07.014 PubMed DOI

Maruyama C, Toyoda J, Kato Y, Izumikawa M, Takagi M, Shin-ya K, Katano H, Utagawa T, Hamano Y (2012) A stand-alone adenylation domain forms amide bonds in streptothricin biosynthesis. Nat Chem Biol 8(9):791–797. https://doi.org/10.1038/nchembio.1040 PubMed DOI

Nah H-J, Pyeon H-R, Kang S-H, Choi S-S, Kim E-S (2017) Cloning and heterologous expression of a large-sized natural product biosynthetic gene cluster in Streptomyces species. Front Microbiol 8:394. https://doi.org/10.3389/fmicb.2017.00394 PubMed DOI PMC

Newman DJ, Cragg GM (2012) Natural products as sources of new drugs over the 30 years from 1981 to 2010. J Nat Prod 75(3):311–335. https://doi.org/10.1021/np200906s PubMed DOI PMC

Pitt JJ (2009) Principles and applications of liquid chromatography-mass spectrometry in clinical biochemistry. Clinic Biochem Rev 30(1):19

Pridham T, Gottlieb D (1948) The utilization of carbon compounds by some Actinomycetales as an aid for species determination. J Bacteriol 56(1):107. https://doi.org/10.1128/JB.56.1.107-114.1948 PubMed DOI PMC

Rajeswari P, Jose PA, Amiya R, Jebakumar SRD (2015) Characterization of saltern based Streptomyces sp. and statistical media optimization for its improved antibacterial activity. Front Microbiol 5:753. https://doi.org/10.3389/fmicb.2014.00753

Reddy T, Mahmood S, Paris L, Reddy YHK, Wellington E, Idris MM (2011) Streptomyces hyderabadensis sp. nov., an actinomycete isolated from soil. Int J System Evolut Microbiol 61 (1):76–80. https://doi.org/10.1099/ijs.0.020446-0

Saadoun I, Muhana A (2008) Optimal production conditions, extraction, partial purification and characterization of inhibitory compound (s) produced by Streptomyces Ds-104 isolate against multi-drug resistant Candida albicans. Curr Trend Biotechnol Pharm 2(2):402–432

Sahana A, Das S, Saha R, Gupta M, Laskar S, Das D (2011) Identification and interaction of amino acids with leucine-anthracene reagent by TLC and spectrophotometry: experimental and theoretical studies. J Chromato Sci 49(8):652–655. https://doi.org/10.1093/chrsci/49.8.652 DOI

Ser HL, Tan WS, Ab Mutalib NS, Yin WF, Chan KG, Goh BH, Lee LH (2016) Draft genome sequence of mangrove-derived Streptomyces sp. MUSC 125 with antioxidant potential. Front Microbiol 7:1470. https://doi.org/10.3389/fmicb.2016.01470

Sharma P, Kalita MC, Thakur D (2016) Broad spectrum antimicrobial activity of forest-derived soil actinomycete, Nocardia sp. PB-52. Front Microbiol 7:347.  https://doi.org/10.3389/fmicb.2016.00347

Shetty PR, Buddana SK, Tatipamula VB, Naga YVV, Ahmad J (2014) Production of polypeptide antibiotic from Streptomyces parvulus and its antibacterial activity. Braz J Microbiol 45(1):303–312. https://doi.org/10.1590/S1517-83822014005000022 PubMed DOI PMC

Shirling ET, Gottlieb D (1966) Methods for characterization of Streptomyces species. Int J System Bacteriol 16(3):313–340. https://doi.org/10.1099/00207713-16-3-313 DOI

Shrivastava P, Kumar R, Yandigeri M (2017) In vitro biocontrol activity of halotolerant Streptomyces aureofaciens K20: a potent antagonist against Macrophomina phaseolina (Tassi) Goid. Saudi J Biol Sci 24(1):192–199. https://doi.org/10.1016/j.sjbs.2015.12.004 PubMed DOI PMC

Singh LS, Sharma H, Talukdar NC (2014) Production of potent antimicrobial agent by actinomycete, Streptomyces sannanensis strain SU118 isolated from phoomdi in Loktak Lake of Manipur. India BMC Microbiol 14(1):278. https://doi.org/10.1186/s12866-014-0278-3 PubMed DOI

Vijayakumar R, Selvam KP, Muthukumar C, Thajuddin N, Panneerselvam A, Saravanamuthu R (2012) Antimicrobial potentiality of a halophilic strain of Streptomyces sp. VPTSA18 isolated from the saltpan environment of Vedaranyam, India. Annal Microbiol 62 (3):1039–1047.  https://doi.org/10.1007/s13213-011-0345-z

Wang X, Huang L, Kang Z, Buchenauer H, Gao X (2010) Optimization of the fermentation process of actinomycete strain Hhs. 015T. J Biomed Biotechnol 2010

Wang Y, Fang X, An F, Wang G, Zhang X (2011) Improvement of antibiotic activity of Xenorhabdus bovienii by medium optimization using response surface methodology. Microb Cell Fact 10(1):98. https://doi.org/10.1155/2010/141876 PubMed DOI PMC

Williams S, Goodfellow M, Wellington E, Vickers J, Alderson G, Sneath P, Sackin M, Mortimer A (1983) A probability matrix for identification of some Streptomycetes. Microbiol 129(6):1815–1830. https://doi.org/10.1099/00221287-129-6-1815 DOI

Xiang W, Liu C, Wang X, Du J, Xi L, Huang Y (2011) Actinoalloteichus nanshanensis sp. nov., isolated from the rhizosphere of a fig tree (Ficus religiosa). Int J System Evol Microbiol 61 (5):1165–1169. https://doi.org/10.1099/ijs.0.023283-0

Yagüe P, López-García MT, Rioseras B, Sánchez J, Manteca A (2013) Pre-sporulation stages of Streptomyces differentiation: state-of-the-art and future perspectives. FEMS Microbiol Lett 342(2):79–88. https://doi.org/10.1111/1574-6968.12128 PubMed DOI