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26 industrialenzimes

  1. 1. World J Microbiol Biotechnol (2009) 25:2103–2111DOI 10.1007/s11274-009-0113-4 ORIGINAL PAPERScreening of marine actinomycetes isolated from the Bayof Bengal, India for antimicrobial activity and industrial enzymesSubramani Ramesh Æ Narayanasamy MathivananReceived: 27 March 2009 / Accepted: 2 July 2009 / Published online: 16 July 2009Ó Springer Science+Business Media B.V. 2009Abstract A total of 288 marine samples were collected marine microorganisms are on the rise, because unique andfrom different locations of the Bay of Bengal starting from biologically active metabolites have been reported fromPulicat lake to Kanyakumari, and 208 isolates of marine marine organisms (Jensen and William 1994; Imada 2004;actinomycetes were isolated using starch casein agar Zhang et al. 2005). Actinomycetes are present in variousmedium. The growth pattern, mycelial coloration, pro- ecological habitats such as soil, fresh water, back water,duction of exopolysaccharides and diffusible pigment and lake, compost, sewage and marine environment (Goodfel-abundance of Streptomyces spp. were documented. Among low and Williams 1983). They are considered highlymarine actinomycetes, Streptomyces spp. were present in valuable as they produce various antibiotics and otherlarge proportion (88%). Among 208 marine actinomycetes, therapeutically useful compounds with diverse biological111 isolates exhibited antimicrobial activity against human activities. The vast majority of these metabolites (70%)pathogens, and 151 showed antifungal activity against two have been isolated from actinomycetes with the remainingplant pathogens. Among 208 isolates, 183, 157, 116, 72 20% from fungi, 7% from Bacillus and 1–2% from Pseu-and 68 isolates produced lipase, caseinase, gelatinase, domonas. Hence, it is known that the actinomycetes arecellulase and amylase, respectively. The results of diver- perhaps the most important group of organisms studiedsity, antimicrobial activity and enzymes production have extensively for the discovery of drugs and other bioactiveincreased the scope of finding industrially important mar- metabolites programme (Lange and Lopez 1996; Prabav-ine actinomycetes from the Bay of Bengal and these athy et al. 2006).organisms could be vital sources for the discovery of Marine environment contains a wide range of distinctindustrially useful molecules/enzymes. microorganisms that are not present in the terrestrial environment. Though some reports are available on anti-Keywords Bay of Bengal Á Actinomycetes Á Diversity Á biotic and enzyme production by marine actinomycetes,Antimicrobial activity Á Extracellular enzymes the marine environment is still a potential source for new actinomycetes, which can yield novel bioactive compounds and industrially important enzymes (Sharma and PantIntroduction 2001). Since late 1980s, the number of novel compounds isolated from terrestrial microorganisms has steadilyMarine microorganisms are increasingly becoming an decreased. To cope up with the demand for new pharma-important source in the search for industrially important ceutical compounds and to combat the antibiotic resistantmolecules. Today both academic and industrial interest in pathogens, researchers have been forced to look for novel microorganisms in unusual environment. Relatively, the Bay of Bengal, an arm of the Indian Ocean has rarely beenS. Ramesh Á N. Mathivanan (&) explored for microbial diversity and microbial metabolites.Biocontrol and Microbial Metabolites Lab, Centre for Advanced Hence, there is an immense possibility to identify newStudies in Botany, University of Madras, Guindy Campus,Chennai, Tamil Nadu 600025, India marine actinomycetes in the Bay of Bengal to discovere-mail: prabhamathi@yahoo.com novel bioactive compounds. Accordingly, the present study 123
  2. 2. 2104 World J Microbiol Biotechnol (2009) 25:2103–2111was aimed to investigate the diversity of industrially Measurement of pHimportant marine actinomycetes in the Bay of Bengal withthe ultimate objective of discovering novel bioactive The pH of water samples was measured directly. Tencompounds. grams of each marine sediment sample was suspended in 20 ml of distilled water. It was allowed to stand for 20 min with intermittent stirring to reach equilibrium. After beingMaterials and methods left to settle, the pH was measured.Study area Isolation of marine actinomycetesThe study area covered the Bay of Bengal coast of Tamil All the marine sediment and seawater samples were sub-Nadu starting from Pulicat lagoon in the north to Kan- jected to pre-heat treatment prior to serial dilution. Pre-heatyakumari in the south. This vast area has a variety of niches treatment was performed by incubating the seawater andsuch as Pulicat lake, Ennore creek, Chennai harbour and sediment samples in a water bath at 50°C for 60 minseveral estuaries viz., Coovum, Adyar, Palar, Vellar, etc. (Takizawa et al. 1993). Ten grams of sediment samplesPulicat lake is the second largest brackish water lagoon in were suspended in 95 ml of sterile aged seawater and theseIndia which runs parallel to the Bay of Bengal. It is located suspensions were considered as 10-1 dilution. Ten milli-at 60 km north-east of Chennai and is separated from the litre of seawater sample was suspended in 90 ml of sterileBay of Bengal by Sriharikota island in Andhra Pradesh aged seawater and these suspensions were considered asstate. The lake is about 360 km2 in size and its depth (water 10-1 dilution. Starch casein agar (SCA) medium containedcolumn) varies from 1 to 6 m. To our knowledge, there is soluble starch 10 g, vitamin free casein 0.3 g, KNO3 2 g,no report from this lake for microbial diversity. Pichavaram NaCl 2 g, K2HPO4 2 g, MgSO4. 7H2O 0.05 g, CaCO3mangrove is located along the coast of Bay of Bengal with 0.02 g, FeSO4Á7H2O 0.01 g, agar 20 g, natural aged sea-11°220 N to 11°300 N wide and 79°450 E to 79°520 E long. water 1,000 ml, pH 7.0 ± 0.2 was used for isolation ofThe total area of this mangrove is about 1,470 ha con- marine actinomycetes. Serial dilutions were done andsisting of about 50 small islands. overlaid on the surface of SCA. The medium also con- tained cycloheximide at 50 lg/ml to minimize fungalCollection of samples contamination. All the plates were incubated at room temperature (28 ± 2°C) for 21 days.A total of 80 seashore sediments, 43 seawater samples, 6 The appearance and growth of marine actinomycetesmarine animals and 5 marine algal samples were collected were observed everyday on SCA plates and the coloniesfrom different locations in Tamil Nadu coast of the Bay of were recognized by their characteristic chalky to leatheryBengal. Totally 21 mangrove sediments, 6 mangrove rhi- appearance. Further, they were observed using a lightzosphere sediments and 10 water samples were collected microscope for their filamentous nature, spores, width offrom Pichavaram and Ennore mangroves in Tamil Nadu. hyphae and spiral sporophores. Individual colonies wereAt least half a kilometer of distance was maintained picked up, and subcultured on SCA and Internationalbetween the sampling stations. The sediment samples were Streptomyces Project medium 2 (ISP2) to ascertain theircollected at 2–3 m depth using grab sampler. Forty-one purity. Various colony characteristics such as size, shape,sediments and two brackish water samples were collected mycelial colour, exopolysaccharide (EPS) and diffusiblefrom Pulicat lake, Tamil Nadu at 5–7 m depth by grab pigment production were recorded. The pure cultures ofsampler. In addition, 26 deep sea sediment samples, 4 corer marine actinomycetes were sub-cultured in SCA slants;sediments and 39 deep seawater samples were collected incubated at room temperature for 5–7 days to achievefrom the Bay of Bengal. The deep sea marine samples were good sporulation; and then preserved in 20% glycerol vialscollected during the cruise programme organized by the at -80°C (Williams and Cross 1971).National Institute of Ocean Technology (NIOT), Chennai,India. A total of four sediments and one water samples Identification of actinomyceteswere collected from the estuary of Adayar river and Marinabeach, Chennai, Tamil Nadu. The sediments and water The cover slip culture technique (Williams and Wilkinssamples were collected in sterile polypropylene bags and 1994) was used to study the morphological characteristicsscrew cap bottles, respectively. The collected samples were such as substrate and aerial mycelia, spores in chain andbrought to the laboratory for isolation of marine actino- chain with coil formation, formation of rectiflexibiles, re-mycetes and the location, nature of sample and pH were tinaculipetri and spiral spores for all the isolates. In addi-documented. tion, nature of Gram’s staining, motility and cell wall123
  3. 3. World J Microbiol Biotechnol (2009) 25:2103–2111 2105amino acid analysis (Becker et al. 1964) were also deter- Each actinomycete strain was streaked on the four cornersmined to identify the marine actinomycetes isolates up to of the respective substrates such as starch, carboxyl methylgenus level. cellulose, gelatin, casein and tween 20 amended agar plates separately and was incubated for 5 days at room temper-Growth characteristics of marine actinomycetes ature. Then the plate was flooded with relevant indicator solution and the development of clear zone around theAll the isolates of marine actinomycetes were grown on growth of organism was considered positive for enzymeSCA at room temperature and the growth rate was moni- activity.tored every day up to 21 days. The isolates, which showedgood growth in 4 days were considered as fast growers andthose that showed good growth between 4 and 7 days were Results and discussionclassified as moderate growers and the slow growers tookmore than 7 days for their growth. In addition to growth, Actinomycetes constantly hold a special significance in themycelial colour was monitored in all the isolates of marine research arena for the past 60 years as the members of thisactinomycetes and documented. group, especially streptomycetes, are known to produce a vast array of compounds with diverse biological properties.Screening of marine actinomycetes for antimicrobial The discovery of new bioactive compounds is a neveractivity ending process to meet the everlasting demand for novel drug and other biomolecules with antimicrobial and ther-All the 208 marine actinomycete isolates were screened for apeutic properties in order to combat human and plantantibacterial and antifungal activity by cross streak and pathogens and also to treat other human ailments. In thisdual culture, respectively. In cross streak method, the scenario, it is more important to identify newer or raremarine isolates were streaked on modified nutrient agar actinomycetes because they are the pivotal sources of(NA) (50% NA ? 50% SCA) as a straight line in the left potent molecules. Marine environment is the biggest res-side corner of the Petriplate and were incubated at room ervoir of chemical and biological diversity. Therefore,temperature for 5 days. After incubation, the test human research focus on marine environment has been gainingbacterial pathogens (Staphylococcus epidermidis importance in recent years. However, still it has not beenMTCC3615, Bacillus subtilis MTCC441, Pseudomonas fully explored and there is tremendous potential to identifyaeruginosa MTCC1688, Escherichia coli MTCC1687 and novel organisms with various biological properties. In lineCandida albicans MTCC227) were streaked at right angle with this view, the present research has been initiated toto the original streak of the actinomycetes isolates. The identify novel actinomycetes from Indian marine environ-zone of inhibition (ZOI) against human bacterial pathogens ment, because its rich microbial diversity has been studiedwas measured after 48 h of incubation. Plates with the only to a limited extent. Totally 288 different marinesame medium without inoculation of actinomycetes but samples were collected from various locations of the Baywith simultaneous streaking of test organisms were main- of Bengal, India (Fig. 1). Among them, a total of 98 acti-tained for controls. nomycetes were isolated from marine sediments, five from Four marine actinomycete isolates were streaked on seawater and nine from marine animals (star fishes, mol-modified potato dextrose agar (PDA) (50% PDA ? 50% lusks and sea urchins), but no actinomycetes were isolatedSCA) as straight lines in four corners of the Petriplate and from marine algae. From Pulicat lake samples, 30 actino-incubated at room temperature for 5 days. After incuba- mycetes were isolated from sediments and 13 fromtion, a fresh mycelial disc of fungal phytopathogens (Rhi- brackish water. From mangrove, 15 actinomycetes werezoctonia solani and Alternaria alternata) was placed in the isolated from sediments, 7 from mangrove rhizospherescenter of each Petriplate and the ZOI against phytopatho- and none of the strains were isolated from mangrove water.gens was measured after 5 days of incubation. The myce- In addition, 18 actinomycete strains were isolated fromlial discs of the test phytopathogens were also kept in deep sea sediments and one strain was isolated from deepcontrol plates where no actinomycetes were inoculated. seawater and corer sediments, respectively. A total of seven actinomycetes were isolated from estuary sedimentsScreening of marine actinomycetes for extracellular and four from estuary water samples (Table 1). Althoughenzymes production soils are considered excellent sources for the isolation of actinomycetes with diverse potential (Ouhdouch et al.All the isolated marine actinomycetes were screened 2001; Lee and Hwang 2002; Prabavathy 2005; Malarvizhiqualitatively for the production of five important enzymes 2006), several actinomycetes have been isolated fromsuch as lipase, caseinase, gelatinase, cellulase and amylase. marine samples (Sujatha et al. 2005; Maldonado et al. 123
  4. 4. 2106 World J Microbiol Biotechnol (2009) 25:2103–2111Fig. 1 Sampling locations in the Bay of Bengal, IndiaTable 1 Details of the isolation of marine actinomycetes from the marine habitats. As actinomycetes represent a small com-Bay of Bengal, India ponent of the total bacterial population in marine sedimentsLocation Nature of sample No. of No. of (Goodfellow and Williams 1983), their role in the marine sample actinomycetes environment is difficult to assess. Nevertheless, these isolate(s) marine actinomycetes are considered economicallyCoastal Sediments 80 98 important as often they are reported to produce valuable Seawater 43 5 bioactive molecules and industrially important enzymes Macro algae 5 0 (Jensen et al. 2005b; Ramesh et al. 2009). Animals 6 9 In this study, 208 different actinomycetes were isolatedDeep sea Sediments 26 18 from 288 marine samples using SCA selective medium Seawater 39 1 prepared in aged seawater. It has already been reported that Corer sediments 4 1 the aged seawater amended media were used to isolate andMangrove Rhizosphere sediments 6 7 maintain the marine microorganisms. Although, a number Sediments 21 15 of selective media (Kuster and Williams 1964; Hayakawa and Nonomura 1987; Crawford et al. 1993; Duangmal et al. Water 10 0 2005; Jensen et al. 2005a) were developed for isolation ofEstuary Sediments 4 7 actinomycetes, SCA was selected, because in this medium Water 1 4 the development of bacterial and fungal colony was veryBrackish lake Sediments 41 30 much suppressed, allowing only the actinomycetes to grow. Brackish water 2 13 Distribution of actinomycetes is influenced by the pH ofTotal 288 208 the respective environment. In the present study, among the 208 marine actinomycetes, 99 isolates were isolated in the2005; Fenical and Jensen 2006; Ramesh et al. 2006, 2009). pH between 8.1 and 8.5, which was followed by the pHThe isolation of actinomycetes from marine sediments was range of 7.6–8.0 from which 92 isolates were obtained.well documented, yet the proportion of these filamentous However, only eight marine actinomycetes were isolatedbacteria which represents the indigenous marine microflora from the pH range of 7.0–7.5 (Table 2). This is in agree-remains unclear. This question persists, in part, because ment with the findings of Taber (1960) who demonstratedthere is little published information describing the distri- that most actinomycetes prefer neutral or slightly alkalinebution, growth and ecological role of actinomycetes in soils for their growth. Similar to the present results,123
  5. 5. World J Microbiol Biotechnol (2009) 25:2103–2111 2107Table 2 Potential of marine pH range Total No. of isolates with antimicrobial activity No. of isolates withactinomycetes isolated from isolates extracellular enzymesdifferent pH on antimicrobial Against human pathogens Against plant pathogensactivity and extracellularenzyme production 7.0–7.5 8 3 6 8 7.6–8.0 92 52 76 88 8.1–8.5 99 50 88 96 Total 199 105 170 192 250 isolates (72.59%) showed fast growth, 47 (22.59%) exhibited slow growth and the remaining of 10 isolates 200 (4.8%) showed moderate growth (Fig. 2). The results Number of isolates clearly revealed that all the marine actinomycetes are not 150 slow growing microorganisms and most of their growth is comparable with filamentous fungi. 100 Among 208 isolates, 115, 79, 6, 7 and 1 were grey, white, blue, pink and orange pigmented, respectively. 50 Interestingly, grey and white mycelial pigmented marine actinomycetes were prominent in the Bay of Bengal. Fur- 0 ther, out of 208 isolates, 6 produced diffusible pigment on Total isolates Fast Moderate Slow SCA agar and 58 isolates produced EPS (Fig. 3). These Growth rate pigments and EPS production could be protective mecha-Fig. 2 Growth rate of marine actinomycetes isolated from the Bay of nisms for actinomycetes to survive in the hostile marineBengal environment. Ramesh et al. (2006) isolated many EPS producing and pigmented actinomycetes in the post-Tsu- nami periods from the Bay of Bengal and most of themMalarvizhi (2006) obtained 33% of the actinomycetes from were able to survive for a long period in marine environ-soils with pH between 7.6 and 8.5. In contrast, Lee and ment compared to non-pigmented and non-EPS producingHwang (2002) isolated many streptomycetes even from actinomycetes. The adaptation of marine microorganismsacidic soil with pH less than 5.0. They also reported the to the diverse marine habitats provides seemingly limitlessdistribution of other actinomycetes genera such as Micro- evolutionary opportunities for the production of uniquemonospora, Dactylosporangium, Streptosporangium, Acti- secondary metabolites.nomadura and Nocardioformis in soils at pH 4.0–5.0. The colonies of actinomycetes were elevated, convexFurther, they observed that Streptomyces were predominant and powdery in nature. Many of such morphologicalin soils with a pH range of 5.1–6.5. characteristics are common in most of the streptomycetes All the isolated marine actinomycetes were screened for (Anderson and Wellington 2001; Lo et al. 2002; Fguiragrowth rate on SCA. Surprisingly, among 208 isolates, 151 et al. 2005; Sujatha et al. 2005). Most of the marineFig. 3 Mycelial colouration of 6 58marine actinomycetes isolatedfrom the Bay of Bengal 7 208 79 6 115 1 Total isolates Orange Grey Blue White Pink EPS Diffusible pigment 123
  6. 6. 2108 World J Microbiol Biotechnol (2009) 25:2103–2111 25 has been reported that the streptomycetes are common inhabitants of marine environments (Kokare et al. 2004a, b; Fiedler et al. 2005; Ramesh et al. 2006), though other actinomycetes are also present (Jensen et al. 1991; Mincer et al. 2002; Magarvey et al. 2004; Maldonado et al. 2008). 208 The degree of antimicrobial activity varied greatly183 among the actinomycetes as shown in Figs. 5 and 6. Among 208 isolates, 111 isolates (53%) showed high Total isolates Streptomycetes Others antimicrobial activity against human pathogens (Fig. 5). Of which, 31, 18, 52, 81, 28 isolates exhibited antimicrobialFig. 4 Abundance of streptomycetes in the Bay of Bengal activity against E. coli, P. aeruginosa, S. epidermidis, B. subtilis and C. albicans, respectively. On the other hand,actinomycetes exhibited different mycelial colourations. four isolates showed antimicrobial activity against all ofThe spore morphology is considered as one of the impor- the five pathogens, but 97 isolates did not show antimi-tant characteristics in the identification of Streptomyces and crobial activity. Several researchers have already reportedit greatly varies among the species (Tresner et al. 1961). It similar antimicrobial activity of actinomycetes againsthas been found that the majority of the marine isolates various human pathogens. Saadoun and Gharaibeh (2003)produced aerial coiled mycelia and the spores arranged in obtained 90 different Streptomyces isolates, of which, 54%chains as already reported by Mukherjee and Sen (2004) exhibited remarkable antibacterial activity against B. sub-and Roes and Meyer (2005). Further, results of the tests as tilis, S. aureus, E. coli, Klebsiella sp. and Shigella sp.outlined in the Bergey’s Manual of Determinative Bacte- Deshmukh and Sridhar (2002) isolated several actinomy-riology (Williams et al. 1989) and the Laboratory Manual cetes from freshwater coastal stream, of which, four iso-for Identification of Actinomycetes (IMTECH 1998) lates inhibited B. subtilis and E. coli. In addition, Imadashowed that among the 208 actinomycetes isolated from et al. (2007) isolated 100 actinomycete strains from variousthe Bay of Bengal, 183 isolates were identified as Strep- locations of the Otsuchi Bay and found that 59 strainstomyces spp. and the remaining 25 isolates belonged to produced antibacterial activity.other genera. Remarkably, majority of the marine actino- In the present study it was recorded that 151 isolatesmycetes (87.98%) were Streptomyces spp. (Fig. 4). The (72%) showed antifungal activity against plant pathogens.cell wall composition is an important criterion for the 107 and 98 isolates exhibited antifungal activity againstidentification of Streptomyces (Sujatha et al. 2005) and R. solani and A. alternata, respectively. On the other hand,chemotaxonomic investigation using isomeric diamino- 54 isolates showed antifungal activity against both thepimelic acid (DAP) configuration was already established fungal phytopathogens, but 57 isolates did not exhibit(Becker et al. 1964; Lechevalier and Lechevalier 1970). It antifungal activity (Fig. 6). Similarly, Yuan and CrawfordFig. 5 Antimicrobial activity of 97marine actinomycetes againsthuman pathogens 4 208 28 81 52 18 111 31 Total isolates Total antimicrobial activity E. coli P. aeruginosa S. epidermidis B. subtilis C. albicans Activity against five pathogens No activity123
  7. 7. World J Microbiol Biotechnol (2009) 25:2103–2111 2109 57 actinomycetes exhibited antimicrobial activity against 54 208 human and plant pathogens, respectively. Among eight isolates from the pH range 7–7.5, 3 (37.5%) and 8 (100%)98 marine actinomycetes showed antimicrobial activity against human and plant pathogens, respectively (Table 2). With the growing awareness on environmental protec- tion, the use of enzymes, particularly from extremophiles, 107 151 gained considerable attention in many industrial processes. In recent years, the microbial enzymes have been replacing Total isolates Total antifungal activity chemical catalysts in manufacturing chemicals, textiles, R. solani A. alternata pharmaceuticals, paper, food and agricultural chemicals. Activity against two pathogens No activity Enzyme-based industrial bioprocess now directly competes with established chemical-based process within the pro-Fig. 6 Antifungal activity of marine actinomycetes against cessed foods, pharmaceutical and allied fermentationphytopathogens industries. In the case of terrestrial actinomycetes, many researchers reported the production of various industrial(1995) demonstrated in vitro antagonism of Streptomyces enzymes (Mohamedin 1999; Azeredo et al. 2001; Pandharelydicus against various fungal phytopathogens in plate et al. 2002; Stamford et al. 2002; Goshev et al. 2005;assay. Further, they observed the inhibition of mycelial Sharma et al. 2005). But to date, it has been concluded thatgrowth in Pythium ultimum and R. solani when grown in there are not many reports on the extracellular enzymesliquid medium with S. lydicus. Kathiresan et al. (2005) from the marine actinomycetes. However, in this study,have isolated 160 actinomycetes from various mangrove among the 208 isolates, 183, 157, 116, 72, 68 isolatesenvironments in India and demonstrated their antifungal produced lipase, caseinase, gelatinase, cellulase and amy-activity against plant pathogenic fungi. Zaitlin et al. (2004) lase, respectively (Fig. 7). Interestingly, 22 isolates pro-demonstrated the antagonistic activity of Streptomyces duced all the five enzymes. The majority of actinomycetehalstedi and Streptomyces rochei against many phyto- strains isolated from the Bay of Bengal produced lipasepathogenic fungi. Search for novel secondary metabolites followed by caseinase, gelatinase, cellulase and amylase.with diverse biological activity in assorted environment has Similarly, Leon et al. (2007) isolated many actinomycetesgained greater attention in recent years. from marine sediments of the central coast of Peru with Among 99 marine actinomycetes isolated from the pH multi-enzyme activity. Notably, a strain among 208 iso-between 8.1 and 8.5, 50 (50.5%) and 88 (88.9%) isolates lates, identified as Streptomyces fungicidicus MML1614,exhibited antimicrobial activity against human and plant was able to produce a thermostable alkaline proteasepathogens, respectively. Among 92 isolates from the pH (Ramesh et al. 2009). These results indicated the potentialbetween 7.6 and 8.0, 52 (56.5%) and 76 (82.6%) marine of marine actinomycetes from the Bay of Bengal for theFig. 7 Extracellular enzymes 22production in marine 72 208actinomycetes 68 116 183 157 Total isolates Lipase Caseinase Gelatinase Amylase Cellulase Five enzymes 123
  8. 8. 2110 World J Microbiol Biotechnol (2009) 25:2103–2111production of various industrial enzymes. Importantly, Duangmal K, Ward CA, Goodfellow M (2005) Selective isolation ofmajority of the actinomycetes (87.98%) obtained from the members of the Streptomyces violaceoruber clad from soil. Microbiol Lett 245:321–327Bay of Bengal produced lipase. The population of lipase Fenical W, Jensen PR (2006) Developing a new resource for drugproducing actinomycetes is relatively large in marine discovery: marine actinomycete bacteria. Nat Chem Biol 2:environment, because the ocean contains significant 666–673amounts of polymers. Microbes have to produce lipase Fguira L, Fotso S, Ben Ameur-Mehdi R, Mellouli L, Laatsch H (2005) Purification and structure elucidation of antifungal andenzyme to degrade the polymers in order to adapt in the antibacterial activities of newly isolated Streptomyces sp. strainextreme environment. US80. Res Microbiol 156:341–347 Among the 99 isolates obtained from the pH between Fiedler HP, Bruntner C, Bull AT, Ward AC, Goodfellow M, Potterat8.1 and 8.5, 96 (97%) marine actinomycetes exhibited O, Puder C, Mihm G (2005) Marine actinomycetes as a source of novel secondary metabolites. Antonie Van Leeuwenhoek 87:extracellular enzyme production, which is of significance. 37–42Among 92 isolates from the pH between 7.6 and 8.0, 88 Goodfellow M, Williams ST (1983) Ecology of actinomycetes. Annu(95.7%) showed extracellular enzyme production. How- Rev Microbiol 37:189–216ever, all of the eight isolates (100%) obtained from the pH Goshev I, Gousterova A, Vasileva-Tonkova E, Nedkov P (2005) Characterization of the enzyme complexed produced by tworange of 7–7.5 produced extracellular enzyme (Table 2). newly isolated thermophilic actinomycete strains during growth on collagen rich materials. Process Biochem 40:1627–1631 Hayakawa M, Nonomura H (1987) Humic acid vitamin agar, a newConclusion medium for the selective isolation of soil actinomycetes. J Ferment Technol 65:501–509 Imada C (2004) Enzyme inhibitors and other bioactive compoundsMarine actinomycetes are metabolically active more vig- from marine actinomycetes. Antonie von Leewenhoek 87:orously in the marine environment, which leads to the 59–63production of various enzymes and bioactive compounds Imada C, Koseki N, Kamata M, Kobayashi T, Hamada-Sato N (2007) Isolation and characterization of antibacterial substances pro-compared to terrestrial strains. Therefore, it is important to duced by marine actinomycetes in the presence of seawater.understand the marine-derived actinomycetes in ecological Actinomycetologica 21:27–31terms and also as a resource for biotechnology. Our present IMTECH (1998) Laboratory manual for identification of actinomy-study and other reports from our lab (Ramesh 2009; Ra- cetes. Institute of Microbial Technology, Chandigarh, p 94 Jensen P, William F (1994) Strategies for the discovery of secondarymesh et al. 2009) evidently revealed that the Bay of Bengal metabolites from marine bacteria. Ecological perspectives. Annuis a potential source for a wide spectrum of antimicrobial Rev Microbiol 48:559–584and industrial enzyme producing actinomycetes. Moreover, Jensen P, Dwight R, Fenical W (1991) The distribution of actinomy-it can be an imperative resource for bioprospecting novel/ cetes in near-shore tropical marine sediments. Appl Environ Microbiol 57:1102–1108rare Streptomyces spp., which could yield valuable bioac- Jensen PR, Gontang E, Mafnas C, Mincer TJ, Fenical W (2005a)tive molecules. Culturable marine actinomycetes diversity from tropical Pacific ocean sediments. Environ Microbiol 7:1039–1048Acknowledgments We thank the Director, CAS in Botany, Uni- Jensen PR, Mincer TJ, Williams PG, Fenical W (2005b) Marineversity of Madras and the Director, National Institute of Ocean actinomycete diversity and natural product discovery. AntonieTechnology, Chennai for laboratory facilities and organizing the Van Leeuwenhoek 87:43–48Cruise programmes, respectively. Kathiresan K, Balagurunathan R, Masilamani Selvam M (2005) Fungicidal activity of marine actinomycetes against phytopath- ogenic fungi. Indian J Biotechnol 4:271–276 Kokare CR, Mahadik KR, Kadam SS, Chopade BA (2004a) Isolation,References characterization and antimicrobial activity of marine halophilic Actinopolyspora species AH1 from the west coast of India. CurrAnderson AS, Wellington EMH (2001) The taxonomy of Streptomy- Sci 86:593–597 ces and related genera. Int J Syst Evol Microbiol 51:797–814 Kokare CR, Mahadik KR, Kadam SS, Chopade BA (2004b) IsolationAzeredo LAI, Leite SGF, Freire DMG, Benchetrit LC, Coelho RRR of bioactive marine actinomycetes from sediments isolated from (2001) Proteases from actinomycetes interfere in solid media Goa and Maharashtra coastlines (west coast of India). Indian J plate assays of hyaluronidase activity. J Microbiol Methods Mar Sci 33:248–256 45:207–212 Kuster E, Williams S (1964) Selection of media for the isolation ofBecker B, Lechevalier MP, Gordon RE, Lechevalier HA (1964) Streptomyces. Nature 202:928–929 Rapid differentiation between Nocardia and Streptomyces by Lange L, Lopez CS (1996) Microorganisms as a source of biolog- paper chromatography of whole cell hydrolysate. Appl Micro- ically active secondary metabolites. In: Copping LG (ed) Crop biol 12:421–424 protection agents from nature: natural products, analogues. TheCrawford DI, Lynch JM, Whipps JM, Ousley MA (1993) Isolation royal society of chemistry, Cambridge and characterization of actinomycetes antagonistic to a fungal Lechevalier HA, Lechevalier MP (1970) A critical evaluation of the root pathogen. Appl Environ Microbiol 59:3899–3909 genera of aerobic actinomycetes. In: Prauser H (ed) TheDeshmukh MB, Sridhar KR (2002) Distribution and antimicrobial actinomycetes. Gustav F Ischer-Verlag, Jena, pp 393–405 activity of actinomycetes of a fresh water coastal stream. Asian J Lee JY, Hwang BK (2002) Diversity of antifungal actinomycetes in Microbiol Biotech Environ Sci 4:335–340 various vegetative soils of Korea. Can J Microbiol 48:407–417123
  9. 9. World J Microbiol Biotechnol (2009) 25:2103–2111 2111Leon J, Liza L, Soto I, Cuadra D, Patino L, Zerpa R (2007) Bioactives Ramesh S, Jayaprakashvel M, Mathivanan N (2006) Microbial status actinomycetes of marine sediment from the central coast of Peru. in seawater and coastal sediments during pre- and post-tsunami Revi Peru Boil 14:259–270 periods in the Bay of Bengal, India. Mar Ecol 27:198–203Lo CW, Lai NS, Cheah HY, Wong NKI, Ho CC (2002) Asian review Ramesh S, Rajesh M, Mathivanan N (2009) Characterization of a of biodiversity and environmental conservation. http://www. thermostable alkaline protease produced by marine Streptomyces arbc.com.my/pdf/art21julysep02.pdf fungicidicus MML1614. Bioprocess Biosyst Eng. doi: 10.1007/Magarvey NA, Keller JM, Bernan V, Dworkin M, Sherman DH s00449-009-0305-1 (2004) Isolation and characterization of novel marine-derived Roes LM, Meyer PR (2005) Streptomyces pharetrae sp. nov., isolated actinomycete taxa rich in bioactive metabolites. Appl Environ from soil from the semi-arid Karoo region. Syst Appl Microbiol Microbiol 70:7520–7529 28:488–493Malarvizhi K (2006) Biodiversity and antagonistic potential of soil Saadoun I, Gharaibeh R (2003) The Streptomyces flora of Badia actinomycetes from south India: isolation, purification and region of Jordan and its potential as a source of antibiotics active characterization of antimicrobial metabolites produced by against resistant bacteria. J Arid Environ 53:365–371 Streptomyces sp. MML1042. Ph.D. Thesis, University of Sharma SL, Pant A (2001) Crude oil degradation by marine Madras, Chennai, India actinomycetes Rhodococcus sp. Indian J Mar Sci 30:146–150Maldonado LA, Fenical W, Jensen PR, Kauffman CA, Mincer TJ, Sharma AD, Kainth S, Gill PK (2005) Inulinase production using Wrad AC, Bull AT, Goodfellow M (2005) Salinispora arenicola garlic (Allium sativum) powder as a potential substrate in gen. nov., and Salinispora tropica nov., obligate marine Streptomyces sp. J Food Eng 77:1–6 actinomycetes belonging to the family Micromonosporaceae. Stamford TLM, Stamford NP, Coelho LCBB, Araujo JM (2002) Int J Syst Evol Microbiol 55:1759–1766 Production and characterization of a thermostable glucoamylase ´ ´ ´Maldonado LA, Fragoso-Yanez D, Perez-Garcıa A, Rosellon-Druker from Streptosporangium endophyte of maize leaves. Bioresour J, Quintana ET (2008) Actinobacterial diversity from marine Technol 83:105–109 sediments collected in Mexico. Antonie van Leeuwenhoek Sujatha P, Bapi Raju KVVSN, Ramana T (2005) Studies on a new 95:111–120. doi: 10.1007/s10482-008-9294-34 marine Streptomycete BT-408 producing polyketide antibioticMincer TJ, Jensen PR, Kauffman CA, Fenical W (2002) Wide spread SBR-22 effective against methicillin resistant Staphylococcus and persistent populations of a major new marine actinomycete aureus. Microbiol Res 160:119–126 taxon in ocean sediments. Appl Environ Microbiol 68:5005– Taber WA (1960) Evidence for the existence of acid-sensitive 5011 actinomycetes in soil. Can J Microbiol 6:503Mohamedin AH (1999) Isolation and identification and some cultural Takizawa M, Colwell RR, Hill RT (1993) Isolation and diversity of conditions of a protease producing thermophilic Streptomyces actinomycetes in the Chesapeake Bay. Appl Environ Microbiol strain grown on chicken feather as a substrate. Int J Biodeter 59:997–1002 Biodeg 43:13–21 Tresner HD, Davies MC, Backus EJ (1961) Electron microscopy ofMukherjee G, Sen SK (2004) Characterization and identification of Streptomyces spore morphology and its role in species differen- chitinase producing Streptomyces venezulae P10. Indian J Exp tiation. J Bacteriol 81:70–80 Biol 42:541–544 Williams ST, Wilkins (1994) Bergey’s manual of determinativeOuhdouch Y, Barakate M, Finanse C (2001) Actinomycetes of bacteriology, 9th edn. Williams and Wilkins, Baltimore Moroccan habitats: isolation and screening for antifungal Williams ST, Cross T (1971) Actinomycetes, methods in microbiol- activities. Eur J Soil Biol 37:69–74 ogy, vol 4. Academic Press, New YorkPandhare J, Zog K, Deshpande VV (2002) Differential stabilities of Williams ST, Goodfellow M, Alderson G (1989) Genus Streptomyces alkaline protease inhibitors from actinomycetes: effect of various Waksman and Henrici 1943, 399AL. In: Williams ST, Sharpe additives on thermostability. Bioresour Technol 84:165–169 ME, Holt JG (eds) Bergey’s manual of systematic bacteriology,Prabavathy VR (2005) Isolation, purification and characterization of vol 4. Williams and Wilkins, Baltimore, pp 2452–2492 antimicrobial metabolites produced by Streptomyces sp. and Yuan W, Crawford DL (1995) Characterization of Streptomyces evolution against blast and sheath diseases of rice. Ph.D Thesis, lydicus WYEC 108 as a potential biocontrol agent against fungal University of Madras, India root and seed rots. Appl Environ Microbiol 61:3119–3128Prabavathy VR, Mathivanan N, Murugesan K (2006) Control of blast Zaitlin B, Turkington K, Parkinson D, Clayton G (2004) Effects of and sheath blight diseases of rice using antifungal metabolites tillage and inorganic fertilizers on culturable soil actinomycetes produced by Streptomyces sp. PM5. Biol Control 39:313–319 communities and inhibitors of fungi by specific actinomycetes.Ramesh S (2009) Marine actinomycetes diversity in Bay of Bengal, Appl Soil Ecol 26:53–62 India: Isolation and characterization of bioactive compounds Zhang L, An R, Wang J, Sun N, Zhang S, Hu J, Kuai J (2005) from Streptomyces fungicidicus MML1614. Ph. D. thesis, Exploring novel bioactive compounds from marine microbes. University of Madras, Chennai, India Curr Opinion Microbiol 8:276–281 123