A novel hydroxamic acid-containing antibiotic produced by a Saharan soil-living Streptomyces strain A. Yekkour, A. Meklat, Christian Bijani, O. Toumatia, R. Errakhi, A. Lebrihi, Florence Mathieu, Abdelghani Zitouni, Nasserdine Sabaou To cite this version: A. Yekkour, A. Meklat, Christian Bijani, O. Toumatia, R. Errakhi, et al.. A novel hydroxamic acid-containing antibiotic produced by a Saharan soil-living Streptomyces strain. Letters in Applied Microbiology, Wiley, 2015, 60 (6), pp.589-596. 10.1111/lam.12412. hal-01938007 HAL Id: hal-01938007 https://hal.archives-ouvertes.fr/hal-01938007 Submitted on 20 May 2019 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. . . . . A novel hydroxamic acid-containing antibiotic produced by Streptomyces a Saharan soil-living strain 1,2 1 3 1 4 4 5 1 A. Yekkour , A. Meklat , C. Bijani , O. Toumatia , R. Errakhi , A. Lebrihi , F. Mathieu , A. Zitouni 1 and N. Sabaou 1 Laboratoire de Biologie des Systemes Microbiens (LBSM), Ecole Normale Superieure de Kouba, Alger, Algeria 2 Centre de Recherche Polyvalent, Institut National de la Recherche Agronomique d’Algerie, Alger, Algeria 3 Laboratoire de Chimie de Coordination (LCC), CNRS, Universite de Toulouse, UPS, INPT, Toulouse, France 4 Universite Moulay Ismail, Meknes, Morocco 5 Universite de Toulouse, Laboratoire de Genie Chimique UMR 5503 (CNRS/INPT/UPS), INP de Toulouse/ENSAT, Castanet-Tolosan Cedex, France Significance and Impact of the Study: This study presents the isolation of a Streptomyces strain, named WAB9, from a Saharan soil in Algeria. This strain was found to produce a new hydroxamic acid-contain- ing molecule with interesting antimicrobial activities towards various multidrug-resistant micro-organ- isms. Although hydroxamic acid-containing molecules are known to exhibit low toxicities in general, only real evaluations of the toxicity levels could decide on the applications for which this new molecule is potentially most appropriate. Thus, this article provides a new framework of research. Keywords Abstract antimicrobial activity, hydroxamic acid, Streptomyces, structure elucidation, During screening for potentially antimicrobial actinobacteria, a highly taxonomy. antagonistic strain, designated WAB9, was isolated from a Saharan soil of Algeria. A polyphasic approach characterized the strain taxonomically as a Correspondence member of the genus Streptomyces. The strain WAB9 exhibited a broad Nasserdine Sabaou, Laboratoire de Biologie spectrum of antimicrobial activity toward various multidrug-resistant micro- des Systemes Microbiens, Ecole Normale organisms. A PCR-based assay of genomic potential for producing bioactive Superieure de Kouba, Alger 16050, Algeria. E-mail: [email protected] metabolites revealed the presence of PKS-II gene. After 6 days of strain fermentation, one bioactive compound was extracted from the remaining aqueous phase and then purified by HPLC. The chemical structure of the compound was determined by spectroscopic (UV–visible, and 1H and 13C doi:10.1111/lam.12412 NMR) and spectrometric analysis. The compound was identified to be 2-amino-N-(2-amino-3-phenylpropanoyl)-N-hydroxy-3-phenylpropanamide, a novel hydroxamic acid-containing molecule. The pure molecule showed appreciable minimum inhibitory concentration values against a selection of drug-resistant bacteria, filamentous fungi and yeasts. Introduction respect to their effects against pathogenic strains and have led to the discovery of a large number of novel active Actinobacteria are a group of ubiquitous Gram-positive compounds of considerable value (Solecka et al. 2012). bacteria with a percentage of guanine–cytosine higher However, the present context of both a decline in the dis- than 55%, and most of them produce mycelia. Because of covery of new antibiotics and an increase in drug-resis- their ability to provide a broad range of bioactive com- tant pathogens makes the screening for new antibiotic- pounds, these micro-organisms have attracted much producing Streptomyces essential. interest. Members of this group, in particular those One of the strategies for enhancing the likelihood of belonging to the genus Streptomyces, are considered as the obtaining particularly interesting isolates and secondary most important producers of bioactive molecules. There- metabolites is to analyse extreme habitats (Santhanam fore, Streptomyces have been thoroughly investigated with et al. 2012), such as arid Saharan soils. Previous surveys on the ecological distribution of Actinomycetes in Alge- Table 1 Antagonistic properties of the strain WAB9 against various multidrug-resistant micro-organisms rian Saharan soils have already demonstrated their appre- ciable biodiversity (Sabaou et al. 1998) and permitted Target micro-organism new species and new antibiotics to be found (Boubetra (antibiotic resistance pattern*) Activity† (mm) et al. 2013; Meklat et al. 2013). Bacillus subtilis ATCC 6633 (NEO) 39 Hydroxamic acids (bearing the functional group N- Staphylococcus aureus S1 28 hydroxy amide) are one such interesting class of molecule (CAR, GEN, K, NEO, OLE, SPI, VAN) produced by many Streptomyces species (Dimkpa et al. Enterobacter cloacae E13 (AMC, ATM, CEP, 29 2008; Kodani et al. 2013). Their low toxicities in general, CTX, FEP, FOX, GEN, PIP, TCC, TIC, TOB) their weak acid properties and their ability for complex- Escherichia coli E52 (ATM, CAZ, CEP, CTX, 35 ing transition metal ions have resulted in these molecules FEP, GEN, PIP, TIC, TOB) Klebsiella pneumoniae K44 (AMX, AN, CF, 29 receiving considerable attention for broad therapeutic and CRO, CTX, CXM, GEN, K, MZ, SSS, TIC) industrial applications (Vanjari and Pande 2003; Rho Pseudomonas aeruginosa IPA1 (AMX, CAR, 29 et al. 2006; Jahangirian et al. 2011). Among their numer- GEN, NEO, SPI, SSS, VAN) ous properties, hydroxamic acids and derivatives have Salmonella enterica E32 (ATM, CAZ, CEP, 16 been largely reported as effective antibacterial and anti- CTX, FEP, GEM, PIP, TIC, TOB) fungal agents (Li et al. 2003; Pepeljnjak et al. 2005). These Fusarium culmorum (CHX, ITR, NYS, TIZ) 39 activities are connected with the ability of hydroxamic Fusarium moniliforme (AMB, CHX, NYS, TIZ) 29 Fusarium sporotrichoides 20 acids to potently and selectively inhibit a range of (AMB, CHX, ITR, NYS, TIZ, TER) enzymes that are vital for the survival of bacteria, such as Fusarium graminearum (CHX, NYS, TIZ) 25 various metalloproteinases, urease, cyclooxygenases and Fusarium oxysporum f. sp. lini (ITR,TIZ) 25 peroxidase (Muri et al. 2004). F. oxysporum f. sp. albedinis 26 Recently, exploration of microbial antibiotic potential (AMB, CHX, ITR, NYS, TIZ, TER) has focused on the screening of polyketide synthase (PKS) Fusarium proliferatum (CHX, ITR, NYS, TIZ) 22 and nonribosomal peptide synthetase (NRPS) genes. This Fusarium equiseti (CHX, TIZ) 30 Aspergillus carbonarius (CHX, NYS) 33 exploration approach seems applicable for hydroxamic Aspergillus niger (AMB, CHX, NYS, TIZ) 25 acid-containing molecules, since these molecules are Aspergillus flavus (TIZ) 26 already known to be synthesized through NRPS-depen- Aspergillus parasiticus (AMB, CHX, ITR, NYS, TIZ) 20 dent or independent pathways (Barry and Challis 2009; Penicillium glabrum (CHX) 38 Oves-Costales et al. 2009). Penicillium expansum 37 Here, we describe the isolation of a highly antagonistic Umbelopsis ramanniana (CHX, ITR, TIZ, TER) 36 Streptomyces strain, designated WAB9, from a Saharan soil Candida albicans IPA 200 (NYS, ITR, TIZ, CHX, TER) 15 Saccharomyces cerevisiae ATCC 4226 34 sample and its identification by conventional and molecu- lar methods, together with production and structure eluci- AMC, amoxicillin + clavulanic acid; AMX, amoxicillin; AN, amikacin; dation of the corresponding bioactive compound, which ATM, aztreonam; C, chloramphenicol; CAR, carbenicillin; CAZ, ceftazi- was found to be a new hydroxamic acid-containing antibi- dime; CFP, cefpirome; CRO, ceftriaxone; CTX, cefotaxime; CXM, ce- otic. The minimum inhibitory concentrations (MICs) of furoxime; FEP, cefepime; FOX, cefoxitin; GEN, gentamicin; K, kanamycin; MZ, mezlocillin; NEO, neomycin; OLE, oleandomycin PIP, this antibiotic were also evaluated on a set of various mul- piperacillin; SPI; spiramycin; SSS, sulfamide; TCC, ticarcillin + clavulan- tidrug-resistant bacteria, filamentous fungi and yeasts. ic acid; TIC, ticarcillin; TOB, tobramycin; VAN, vancomycin; AMB, amphotericin B; CHX, cycloheximide; ITR, itraconazole; NYS, nystatin; TIZ, thioconazole; TER, terbinafine. Results and discussion *The antibiotic resistance patterns were evaluated in house as The strain WAB9 was isolated by plating a 2 : 10 serial described by Touati et al. (2006). dilution of the soil sample of Bechar on chitin-vitamin †Activity estimated by measuring the length of inhibition between agar medium supplemented with selective antibiotics. Actinomycetes and target micro-organisms. This strain was found in only a single small colony, which shows it rarity. the targeted bacteria, filamentous fungi and