Antibacterial and Antioxidant Activities of Novel Actinobacteria Strain Isolated from Gulf of Khambhat, Gujarat.

Bacterial secondary metabolites possess a wide range of biologically active compounds including antibacterial and antioxidants. In this study, a Gram-positive novel marine Actinobacteria was isolated from sea sediment which showed 84% 16S rRNA gene sequence (KT588655) similarity with Streptomyces variabilis (EU841661) and designated as Streptomyces variabilis RD-5. The genus Streptomyces is considered as a promising source of bioactive secondary metabolites. The isolated novel bacterial strain was characterized by antibacterial characteristics and antioxidant activities. The BIOLOG based analysis suggested that S. variabilis RD-5 utilized a wide range of substrates compared to the reference strain. The result is further supported by statistical analysis such as AWCD (average well color development), heat-map and PCA (principal component analysis). The whole cell fatty acid profiling showed the dominance of iso/anteiso branched C15-C17 long chain fatty acids. The identified strain S. variabilis RD-5 exhibited a broad spectrum of antibacterial activities for the Gram-negative bacteria (Escherichia coli NCIM 2065, Shigella boydii NCIM, Klebsiella pneumoniae, Enterobacter cloacae, Pseudomonas sp. NCIM 2200 and Salmonella enteritidis NCIM), and Gram-positive bacteria (Bacillus subtilis NCIM 2920 and Staphylococcus aureus MTCC 96). Extract of S. variabilis strain RD-5 showed 82.86 and 89% of 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging and metal chelating activity, respectively, at 5.0 mg/mL. While H2O2 scavenging activity was 74.5% at 0.05 mg/mL concentration. Furthermore, polyketide synthases (PKSs types I and II), an enzyme complex that produces polyketides, the encoding gene(s) detected in the strain RD-5 which may probably involve for the synthesis of antibacterial compound(s). In conclusion, a novel bacterial strain of Actinobacteria, isolated from the unexplored sea sediment of Alang, Gulf of Khambhat (Gujarat), India showed promising antibacterial activities. However, fractionation and further characterization of active compounds from S. variabilis RD-5 are needed for their optimum utilization toward antibacterial purposes.

Biological neutralization and biosorption of dyes of alkaline textile industry wastewater.

The present work was aimed to secure biological neutralization and biosorption of dyes of an alkaline textile industrial effluent (ATIE) using an alkaliphilic bacterium, Enterococcus faecalis strain R-16 isolated from Gujarat coast. The isolate was capable and competent to bring down the pH of ATIE from 12.1 to 7.0 within 2 h in the presence of carbon and nitrogen sources. Carboxylic group concentration (CGC), NMR and FT-IR analysis revealed production of carboxylic acid as a result of neutralization. The unconventional carbon and nitrogen sources like Madhuca indica flowers or sugar cane bagasse supported the growth of bacterium with effective neutralization and biosorption of dyes from ATIE. The process proved to be efficient, inexpensive and eco-friendly as compared to conventional chemical neutralization process.

Occurrence and distribution of selected heavy metals and boron in groundwater of the Gulf of Khambhat region, Gujarat, India.

The concentration of selected heavy metals, like As, Co, Cr, Cu, Fe, Mn, Ni, Pb, and Zn as well as B, was measured by inductively coupled plasma-optical emission spectrometry (ICP-OES) in groundwater samples from various locations in the Gulf of Khambhat (GoK), an inlet of the Arabian Sea in the state of Gujarat, India, during post-monsoon, winter, and pre-monsoon seasons in a year. Most heavy elements are characterized by low mobility under slightly alkaline and reducing conditions; concentrations in confined aquifers are smaller than the maximum permissible values for drinking water. The temporal changes indicate that a majority of metals is entering the aquifer during monsoon. Principle component analysis of the heavy metal data suggests that Co, Cu, Cd, and Zn are interrelated with each other and derived significantly from anthropogenic route, while input of Pb and Cr may be due to atmospheric deposition in the study area. Both weathering of rocks and anthropogenic input were found to be main sources of elements in the groundwater. The heavy metal levels in groundwaters of the GoK region in comparison with some of the European and Asian sites were higher; however, these metal levels were found to be comparable with few urban sites in the world.

Effect of unconventional carbon sources on biosurfactant production and its application in bioremediation.

The potential of an alkaliphilic bacterium Klebsiella sp. strain RJ-03, to utilize different unconventional carbon sources for the production of biosurfactant was evaluated. The biosurfactant produced using corn powder, potato peel powder, Madhuca indica and sugarcane bagasse containing medium, exhibited significantly higher viscosity and maximum reduction in surface tension as compared to other substrates. Among several carbon substrates tested, production of biosurfactant was found to be the highest with corn powder (15.40 ± 0.21 g/l) as compared to others. The comparative chemical characterization of purified biosurfactant was done using advance analytical tools such as NMR, FT-IR, SEM, GPC, MALDI TOF-TOF MS, GC-MS, TG and DSC. Analyses indicated variation in the functional groups, monosaccharide composition, molecular mass, thermostability. Higher yield with cheaper raw materials, noteworthy stress tolerance of CP-biosurfactant toward pH and salt as well as compatibility with chemical surfactants and detergents revealed its potential for commercialization and application in bioremediation.

Production and structural characterization of biosurfactant produced by an alkaliphilic bacterium, Klebsiella sp.: evaluation of different carbon sources.

The potential of an alkaliphilic bacterium Klebsiella sp. strain RJ-03, to utilize different carbon sources for the production of an extracellular biosurfactant was evaluated. Among the several carbon substrates tested, production of the crude biosurfactant was found to be the highest with starch (10.1±0.11g/L) followed by sucrose (5.1±0.11g/L), xylose (3.25±0.08g/L), galactose (3.1±0.16g/L) glucose (2.75±0.11g/L) and fructose (2.62±0.07g/L). The crude biosurfactant production was done using starch, sucrose, xylose, galactose and glucose containing medium, that exhibited significantly high viscosity, emulsification activity and maximum reduction in surface tension as compared to those obtained from fructose and maltose. The carbon source has significant effect on the quantity as well as the quality of biosurfactant production. The chemical characteristics of purified biosurfactant was compared by NMR, FT-IR, SEM, GPC, MALDI TOF-TOF MS, GC-MS, TG and DSC analysis, indicating variation in the functional groups, bonds, elements, monosaccharide composition, molecular mass and thermo stability.

Physicochemical characterization of biosurfactant and its potential to remove oil from soil and cotton cloth.

An alkaliphilic bacterium, Klebsiella sp. strain RJ-03, produced a biosurfactant, which showed low viscosity with pseudoplastic rheological behavior and exhibited emulsification activity with oils and hydrocarbons. The biosurfactant has excellent oil removing efficiency as compared to chemical surfactants. The isolated biosurfactant has compatibility with detergents and enhanced oil removing efficiency from soil and cotton cloths. It comprised of sugar, uronic acid, protein and sulfate. GC-MS analysis confirmed the presence of six monosaccharides (w/w), glucose (6.65%), galactose (23.98%), rhamnose (14.94%), mannose (17.54%), fucose (9.47%) and 6-O-Me-galactose (1.4%). It is a high molecular weight, thermostable biopolymer showing degradation above 300 °C. Positive ion reflector mode of MALDI TOF-TOF MS analysis revealed series of low and mid range mass peaks (m/z) corresponding to mono-, di-, tri- and oligo-saccharides content. The NMR, FT-IR, EDX-SEM, AFM and PSD analysis confirmed the presence of functional groups, bonds, elements and particle size respectively.

Kappaphycus alvarezii as a source of bioethanol.

The present study describes production of bio-ethanol from fresh red alga, Kappaphycus alvarezii. It was crushed to expel sap--a biofertilizer--while residual biomass was saccharified at 100 °C in 0.9 N H2SO4. The hydrolysate was repeatedly treated with additional granules to achieve desired reducing sugar concentration. The best yields for saccharification, inclusive of sugar loss in residue, were 26.2% and 30.6% (w/w) at laboratory (250 g) and bench (16 kg) scales, respectively. The hydrolysate was neutralized with lime and the filtrate was desalted by electrodialysis. Saccharomyces cerevisiae (NCIM 3523) was used for ethanol production from this non-traditional bio-resource. Fermentation at laboratory and bench scales converted ca. 80% of reducing sugar into ethanol in near quantitative selectivity. A petrol vehicle was successfully run with E10 gasoline made from the seaweed-based ethanol. Co-production of ethanol and bio-fertilizer from this seaweed may emerge as a promising alternative to land-based bio-ethanol.

Biological neutralization of chlor-alkali industry wastewater.

The present work reports biological neutralization of chlor-alkali industrial effluent by an alkaliphilic bacterium, isolated from the Gujarat coast, which was identified as Enterococcus faecium strain R-5 on the basis of morphological, biochemical and partial 16S rRNA gene sequencing. The isolate was capable of bringing down the pH of waste water from 12.0 to 7.0 within 3 h in the presence of carbon and nitrogen sources, with simultaneous reduction in total dissolved solutes (TDS) up to 19-22%. This bacterium produced carboxylic acid, as revealed by FT-IR analysis, which facilitated neutralization of alkaline effluent. The presence of unconventional raw materials viz. Madhuca indica flowers or sugar cane bagasse as carbon and nitrogen sources could effectively neutralize alkaline effluent and thus making the bioremediation process economically viable. The time required for neutralization varied with size of inoculum. To the best of our knowledge, this is the first report on biological neutralization of a chlor-alkali industrial effluent.

Abundance, diversity and antibiotics resistance pattern of Vibrio spp. in coral ecosystem of Kurusadai island.

The abundance and species diversity of Vibrio associated with coral reef ecosystem of Kurusadai island, Tamil Nadu, India were evaluated. A total of twelve sampling locations including different live and dead coral surfaces, surrounding water and rock surface (negative control) were selected for the present study. Total viable and TCBS counts were found to be higher in dead coral as compared to that of live coral. Out of total 21 species of Vibrio isolated, 13 were identified up to species level based on biochemical tests and 16S rRNA gene sequence homology, while remaining 8 isolates did not show homology up to species level with any of the sequences available in the NCBI database. Moreover, these unidentified Vibrio spp. exhibited intra-species variation. This study indicated association of hitherto unknown Vibrio species with coral reef ecosystem of Kurusadai island. Assuming that only resistant bacteria can grow in the coral environment, susceptibility against a total of 20 antibiotics was evaluated. All the isolates exhibited resistance towards more than 6 antibiotics. Interestingly, none of the identified bacteria were previously reported to be of coral pathogen reflecting the healthy nature of the ecosystem. However, a continuous monitoring of the region will be prerequisite to envisage the role of these bacteria on the health status of the coral ecosystem.

Evaluation of biosurfactant/bioemulsifier production by a marine bacterium.

Planococcus maitriensis Anita I (NCBI GenBank Accession number EF467308) was tested for its biosurfactant/bioemulsifying efficacy. The crude extracellular polymeric substance (EPS) produced by this bacterium contained carbohydrate (12.06%), protein (24.44%), uronic acid (11%) and sulfate (3.03%). The oil spreading potential of this EPS was comparable to Triton X100 and Tween 80. This exopolymer emulsified xylene more efficiently as compared to few standard gums. It also formed stable emulsions (E (1,080)=100) with jatropha, paraffin and silicone oils. The cell free supernatant of this bacterium successfully reduced the surface tension (from 72 to 46.07 mN m(-1)). It also decreased interfacial tension of hexane and xylene. Based on the emulsifying and tensiometric properties, this bacterium or its exopolymer could be used for bioremediation, enhanced oil recovery and in cosmetics.

Bacterial exopolysaccharides--a perception.

Microbial polysaccharides are multifunctional and can be divided into intracellular polysaccharides, structural polysaccharides and extracellular polysaccharides or exopolysaccharides (EPS). Extracellular polymeric substances (EPS), produced by both prokaryotes (eubacteria and archaebacteria) and eukaryotes (phytoplankton, fungi, and algae), have been of topical research interest. Newer approaches are carried out today to replace the traditionally used plant gums by their bacterial counterparts. The bacterial exopolysaccharides represent a wide range of chemical structures, but have not yet acquired appreciable significance. Chemically, EPS are rich in high molecular weight polysaccharides (10 to 30 kDa) and have heteropolymeric composition. They have new-fangled applications due to the unique properties they possess. Owing to this, exopolysaccharides have found multifarious applications in the food, pharmaceutical and other industries. Hence, the present article converges on bacterial exopolysaccharides.

Characterization of an exopolysaccharide produced by a marine Enterobacter cloacae.

An exopolysaccharide producing marine bacterium, Enterobacter cloacae, was isolated from marine sediment collected from Gujarat coast, India. Chemical investigation of exopolysaccharide (EPS 71 a) revealed that this exopolysaccharide was an acidic polysaccliaride containing high amount of uronic acid, fucose and sulfate which is rare for bacterial exopolysaccharides. EPS 71a was found to have fucose, galactose, glucose and glucuronic acid in a molar ratio of 2: 1: 1: 1.

Biosorption of heavy metals by a marine bacterium.

Heavy metal chelation property of exopolysaccharide produced by Enterobacter cloaceae, a marine bacterium, isolated from the West Coast of India, is reported in this paper. The exopolysaccharide demonstrated excellent chelating properties with respect to cadmium (65%) followed by copper (20%) and cobalt (8%) at 100 mg/l heavy metal concentration. However, it could not chelate mercury. A comparative study of the percentage biosorption of the above mentioned metals is presented here.

Accumulation of hexavalent chromium by an exopolysaccharide producing marine Enterobacter cloaceae.

An exopolysaccharide producing Enterobacter cloaceae (AK-I-MB-71a) was tested for its Cr (VI) tolerance. This isolate was not only resistant to this heavy metal but also showed enhanced growth and exopolysaccharide production in the presence of Cr (VI) at 25, 50 and 100 ppm concentrations. XRF analysis of both the biomass as well as the exopolysaccharide revealed that a sum total of about 60-70% chromium was accumulated by this bacterium. This indicated that this organism could prove to be a potential candidate in the field of bioremediation with respect to chromium removal.