Shunmugiah Karutha Pandian

Alagappa University, Karaikudi, Tamil Nādu, India

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Publications (73)162.18 Total impact

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    ABSTRACT: Bacterial urinary tract infections (UTIs) are the most common nosocomial infections, accounting for about 40 % of all hospital-acquired infections. The bacterial spectrum of nosocomial UTIs is broad and the treatment of UTIs is becoming difficult owing to the emergence of drug resistance. Therefore, it is reasonable to investigate novel and alternative therapeutic strategies to treat UTIs. Since UTIs are caused by uropathogens with quorum sensing (QS)-dependent biofilm forming abilities, interruption of QS systems may be a novel approach to combat drug resistance. In the present study, a methanol extract (and hexane extract derived from it) of the medicinal plant Hyptis suaveolens (L.) were shown to have anti-QS activity against the biosensor strain Chromobacterium violaceum (ATCC 12472). Furthermore, the hexane extract of H. suaveolens (HEHS) inhibited biofilm formation by uropathogens such as Escherichia coli, Proteus vulgaris, Proteus mirabilis, Klebsiella pneumoniae and Serratia marcescens. HEHS promotes the loosening of biofilm architecture and strongly inhibits in vitro biofilm formation by uropathogens, which was more apparent from microscopic images. In addition to this, HEHS reduces the production of QS-dependent virulence factors like protease and hemolysin, along with motility. The partial purification and GC-MS analysis of the active fraction revealed the presence of several therapeutically important compounds which may synergistically act on the uropathogens and possibly reduce the QS-dependent phenotypes. These findings suggest HEHS as potential phytotherapeutic agent which can be employed to formulate protective strategies against biofilm linked infections caused by uropathogens.
    Antonie van Leeuwenhoek 02/2015; · 2.07 Impact Factor
  • Alwar Ramanujam Padmavathi, Murugesan Periyasamy, Shunmugiah Karutha Pandian
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    ABSTRACT: Extracellular polymeric substances (EPS) play crucial roles in biofilm formation and biocorrosion resulting in heavy economic loss in an industrial setup. Hence, in an attempt to develop an agent to control the EPS across the hosts, the ability of 2,4-Di-tert-butylphenol (DTBP), a potent antioxidant, to modify the EPS of Serratiamarcescens has been investigated in this study using biophysical methods. Protein, polysaccharides and eDNA components of EPS were inhibited significantly (p<0.05) upon exposure to DTBP. DTBP treatment reduced the crystallite size and crystallinity index of EPS and increased the dislocation density of crystallites without inducing stress, besides increasing the hydration of EPS which reduced its thermal stability. On the whole, this study highlights the efficacy of DTBP to modulate secreted EPS of S. marcescens which in turn could facilitate the disruption of biofilms besides favouring the diffusion of antimicrobials into the cell aggregates resulting eradication of persistent biofilms. Copyright © 2015 Elsevier Ltd. All rights reserved.
    Bioresource Technology 01/2015; · 5.04 Impact Factor
  • Alwar Ramanujam Padmavathi, Shunmugiah Karutha Pandian
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    ABSTRACT: Coral Associated Bacteria (CAB) (N = 22) isolated from the mucus of the coral Acropora digitifera were screened for biosurfactants using classical screening methods; hemolysis test, lipase production, oil displacement, drop collapse test and emulsifying activity. Six CAB (U7, U9, U10, U13, U14, and U16) were found to produce biosurfactants and were identified by 16S ribosomal RNA gene sequencing as Providencia rettgeri, Psychrobacter sp., Bacillus flexus, Bacillus anthracis, Psychrobacter sp., and Bacillus pumilus respectively. Their cell surface hydrophobicity was determined by Microbial adhesion to hydrocarbon assay and the biosurfactants produced were extracted and characterized by Fourier Transform Infrared spectroscopy. Since the biosurfactants are known for their surface modifying capabilities, antibiofilm activity of positive isolates was evaluated against biofilm forming Pseudomonas aeruginosa ATCC10145. Stability of the active principle exhibiting antibiofilm activity was tested through various temperature treatments ranging from 60 to 100 °C and Proteinase K treatment. CAB isolates U7 and U9 exhibited stable antibiofilm activity even after exposure to higher temperatures which is promising for the development of novel antifouling agents for diverse industrial applications. Further, this is the first report on biosurfactant production by a coral symbiont.
    Indian Journal of Microbiology 12/2014; 54(4). · 0.83 Impact Factor
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    ABSTRACT: Group A Streptococci (GAS) are involved in a number of life threatening diseases and biofilm formation by these pathogens are considered as an important virulence determinant as it mediates antibiotic resistance among them. In the present study, we have explored the ability of (+)-usnic acid, a lichen secondary metabolite, as an antibiofilm agent against four serotypes of Streptococcus pyogenes causing pharyngitis. Usnic acid inhibited the biofilms of M serotypes M56, st38, M89 efficiently and the biofilm of M74 to a lesser extent. Confocal imaging of the treated samples showed that usnic acid reduced the biomass of the biofilms when compared to that of the control. Fourier Transfer Infrared (FT-IR) spectroscopy indicated that usnic acid reduced the cellular components (proteins and fatty acids) of the biofilms. Interestingly, the FT-IR spectrum further revealed that usnic acid probably acted upon the fatty acids of the biofilms as evident from the disappearance of a peak at 2,455-2,100 cm(-1) when compared to the control only in serotypes M56, st38 and M89 but not in M74. The present study shows, for the first time, that usnic acid can act as an effective antibiofilm agent against GAS.
    Antonie van Leeuwenhoek 11/2014; · 2.07 Impact Factor
  • Alwar Ramanujam Padmavathi, Bose Abinaya, Shunmugiah Karutha Pandian
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    ABSTRACT: Intercellular communication in bacteria (quorum sensing, QS) is an important phenomenon in disease dissemination and pathogenesis, which controls biofilm formation also. This study reports the anti-QS and anti-biofilm efficacy of seaweed Gracilaria gracilis associated Vibrio alginolyticus G16 against Serratia marcescens. Purification and mass spectrometric analysis revealed the active principle as phenol, 2,4-bis(1,1-dimethylethyl) [PD]. PD affected the QS regulated virulence factor production in S. marcescens and resulted in a significant (p < 0.05) reduction in biofilm (85%), protease (41.9%), haemolysin (69.9%), lipase (84.3%), prodigiosin (84.5%) and extracellular polysaccharide (84.62%) secretion without hampering growth, as evidenced by XTT [2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide] assay. qPCR analysis confirmed the down-regulation of the fimA, fimC, flhD and bsmA genes involved in biofilm formation. Apart from biofilm inhibition and disruption, PD increased the susceptibility of S. marcescens to gentamicin when administered synergistically, which opens another avenue for combinatorial therapy where PD can be used to enhance the efficacy of conventional antibiotics.
    Biofouling 10/2014; 30(9):1111-22. · 3.40 Impact Factor
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    ABSTRACT: The recognition of DNA by small molecules is of special importance in the design of new drugs. Many natural and synthetic compounds have the ability to interact with the minor groove of DNA. In the present study, identification of minor groove binding compounds was attained by the combined approach of pharmacophore modelling, virtual screening and molecular dynamics approach. Experimentally reported 32 minor groove binding compounds were used to develop the pharmacophore model. Based on the fitness score, best three pharmacophore hypotheses were selected and used as template for screening the compounds from drug bank database. This pharmacophore-based screening provides many compounds with the same pharmacological properties. All these compounds were subjected to four phases of docking protocols with combined Glide-quantum-polarized ligand docking approach. Molecular dynamics results indicated that selected compounds are more active and showed good interaction in the binding site of DNA. Based on the scoring parameters and energy values, the best compounds were selected, and antibacterial activity of these compounds was identified using in vitro antimicrobial techniques. Copyright © 2014 John Wiley & Sons, Ltd.
    Journal of Molecular Recognition 07/2014; 27(7). · 3.01 Impact Factor
  • Shanmugaraj Gowrishankar, Balan Poornima, Shunmugiah Karutha Pandian
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    ABSTRACT: Since Streptococcus mutans is the principal etiologic agent causing dental caries, by encompassing an array of unique virulence traits, emerging treatment strategies that specifically target the virulence of this pathogen may be promising as alternative approaches compared to conventional antibiotic therapy. In this perspective, we investigated chloroform extract of cell-free culture supernatant from mangrove rhizosphere bacterium Bacillus amyloliquefaciens (MMS-50) in terms of anticariogenic properties of S. mutans, without suppressing its viability. Crude chloroform extract of MMS-50 was subjected to column and high performance liquid chromatographic techniques to obtain the active fraction (AF), and MMS-50 AF was used for all further assays. GC–MS and FT-IR were carried out to identify the major components present in MMS-50 AF. Comparative gene expression analysis of some biofilm-forming and virulence genes (vicR, comDE, gtfC, and gbpB) was done by real-time PCR. Cyclo(L-leucyl-L-prolyl) was found to be the chief compound in MMS-50 AF responsible for bioactivity. The minimum and maximum inhibitory concentrations of MMS-50 AF against S. mutans were found to be 100 and 250 μg/mL, respectively. Anti-virulence assays performed using below-sub-MIC levels of MMS-50 AF (30 μg/mL) resulted in significant reduction in adherence (68%), acid production, acid tolerance, glucan synthesis (32%), biofilm formation (53.5%) and cell surface hydrophobicity, all devoid of affecting its viability. The micrographs of CLSM and SEM further confirmed the antibiofilm and anti-virulence efficacies of MMS-50 AF. Expression data showed significant reduction in expression of all studied virulence genes. Thus, the current study unveils the anticariogenic potential of cyclo(L-leucyl-L-prolyl) from B. amyloliquefaciens, as well as its suitability as a novel and alternative anticariogenic agent against dental caries.
    Research in Microbiology 05/2014; · 2.83 Impact Factor
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    ABSTRACT: Urinary tract infection is caused primarily by the quorum sensing (QS)-dependent biofilm forming ability of uropathogens. In the present investigation, an anti-quorum sensing (anti-QS) agent curcumin from Curcuma longa (turmeric) was shown to inhibit the biofilm formation of uropathogens, such as Escherichia coli, Pseudomonas aeruginosa PAO1, Proteus mirabilis and Serratia marcescens, possibly by interfering with their QS systems. The antibiofilm potential of curcumin on uropathogens as well as its efficacy in disturbing the mature biofilms was examined under light microscope and confocal laser scanning microscope. The treatment with curcumin was also found to attenuate the QS-dependent factors, such as exopolysaccharide production, alginate production, swimming and swarming motility of uropathogens. Furthermore, it was documented that curcumin enhanced the susceptibility of a marker strain and uropathogens to conventional antibiotics.
    Food Chemistry 04/2014; 148C:453-460. · 3.26 Impact Factor
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    ABSTRACT: The LuxS-based signalling pathway has an important role in physiological and pathogenic functions that are capable of causing different infections. In the present study, cinnamaldehyde (CN) and their derivatives were evaluated for their inhibitory efficiency against LuxS by molecular modelling, docking, dynamics and free-energy calculations. Sequence and structure-similarity analysis of LuxS protein, five different amino acids were found to be highly conserved, of which GLY128 was identified as the key residue involved in the effective binding of the ligands. Quantum-polarized ligand docking protocol showed that 2nitro and 4nitro CN has a higher binding efficiency than CN, which very well corroborates with the in vitro studies. COMSTAT analysis for the microscopic images of the S. pyogenes biofilm showed that the ligands have antibiofilm potential. In addition, the results of quantitative polymerase chain reaction (qPCR) analysis revealed that the transcripts treated with the compounds showed decrease in luxS expression, which directly reflects with the reduction in expression of speB. No substantial effect was observed on the virulence regulator (srv) transcript. These results confirm that speB is controlled by the regulation of luxS. The decreased rate of S. pyogenes survival in the presence of these ligands envisaged the fact that the compounds could readily enhance opsonophagocytosis with the reduction of virulence factor secretion. Thus, the overall data supports the use of CN derivatives against quorum sensing-mediated infections caused by S. pyogenes. Copyright © 2014 John Wiley & Sons, Ltd.
    Journal of Molecular Recognition 02/2014; 27(2):106-16. · 3.01 Impact Factor
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    ABSTRACT: Bacterial community composition in the marine environment differs from one geographical location to another. Reports that delineate the bacterial diversity of different marine samples from geographically similar location are limited. The present study aims to understand whether the bacterial community compositions from different marine samples harbour similar bacterial diversity since these are geographically related to each other. In the present study, 16S rRNA deep sequencing analysis targeting V3 region was performed using Illumina bar coded sequencing. A total of 22.44 million paired end reads were obtained from the metagenomic DNA of Marine sediment, Rhizosphere sediment, Seawater and the epibacterial DNA of Seaweed and Seagrass. Diversity index analysis revealed that Marine sediment has the highest bacterial diversity and the least bacterial diversity was observed in Rhizosphere sediment. Proteobacteria, Actinobacteria and Bacteroidetes were the dominant taxa present in all the marine samples. Nearly 62-71% of rare species were identified in all the samples and most of these rare species were unique to a particular sample. Further taxonomic assignment at the phylum and genus level revealed that the bacterial community compositions differ among the samples. This is the first report that supports the fact that, bacterial community composition is specific for specific samples irrespective of its similar geographical location. Existence of specific bacterial community for each sample may drive overall difference in bacterial structural composition of each sample. Further studies like whole metagenomic sequencing will throw more insights to the key stone players and its interconnecting metabolic pathways. In addition, this is one of the very few reports that depicts the unexplored bacterial diversity of marine samples (Marine sediment, Rhizosphere sediment, Seawater) and the host associated marine samples (Seaweed and Seagrass) at higher depths from uncharacterised coastal region of Palk Bay, India using next generation sequencing technology.
    PLoS ONE 10/2013; 8(10):e76724. · 3.53 Impact Factor
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    ABSTRACT: Computational analysis of sequence homology of the chiSRC gene cluster, encoding a chitinase in Streptomyces peucetius, showed that the gene cluster could be a two-component regulon comprising a sensor kinase (chiS) and a response regulator (chiR). To prove that the ChiSRC is an authentic two-component system, the chiS gene was cloned and expressed in E.coli and the purified protein was used for biochemical analysis. In this report, we provide biochemical evidence to show that the sensor kinase encoded by chiS gene indeed is a histidine kinase capable of autophosphorylation and the histidine 144 residue of the ChiS protein is the phosphate acceptor. An insertion mutation at the chiS locus led to overproduction chitinase protein in S. peucetius implying that the chiC gene is negatively regulated by the two-component system.
    Microbiological Research 08/2013; · 1.94 Impact Factor
  • Rajamohmed Beema Shafreen, Shunmugiah Karutha Pandian
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    ABSTRACT: Streptococcus pyogenes (SP) is the major cause of pharyngitis accompanied by strep throat infections in humans. 3-keto acyl reductase (FabG), an important enzyme involved in the elongation cycle of the fatty acid pathway of S. pyogenes, is essential for synthesis of the cell-membrane, virulence factors and quorum sensing-related mechanisms. Targeting SPFabG may provide an important aid for the development of drugs against S. pyogenes. However, the absence of a crystal structure for FabG of S. pyogenes limits the development of structure-based drug designs. Hence, in the present study, a homology model of FabG was generated using the X-ray crystallographic structure of Aquifex aeolicus (PDB ID: 2PNF). The modeled structure was refined using energy minimization. Furthermore, active sites were predicted, and a large dataset of compounds was screened against SPFabG. The ligands were docked using the LigandFit module that is available from Discovery Studio version 2.5. From this list, 13 best hit ligands were chosen based on the docking score and binding energy. All of the 13 ligands were screened for Absorption, Distribution, Metabolism, Excretion and Toxicity (ADMET) properties. From this, the two best descriptors, along with one descriptor that lay outside the ADMET plot, were selected for molecular dynamic (MD) simulation. In vitro testing of the ligands using biological assays further substantiated the efficacy of the ligands that were screened based on the in silico methods.
    Journal of molecular graphics & modelling 08/2013; 45C:1-12. · 2.17 Impact Factor
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    ABSTRACT: Infectious diseases caused by bacteria and fungi are the major cause of morbidity and mortality across the globe. Multi-drug resistance in these pathogens augments the complexity and severity of the diseases. Various studies have shown the role of biofilms in multi-drug resistance, where the pathogen resides inside a protective coat made of extracellular polymeric substances. Since biofilms directly influence the virulence and pathogenicity of a pathogen, it is optimal to employ a strategy that effectively inhibits the formation of biofilm. Pomegranate is a common food and is also used traditionally to treat various ailments. This study assessed the anti-biofilm activity of a methanolic extract of pomegranate against bacterial and fungal pathogens. Methanolic extract of pomegranate was shown to inhibit the formation of biofilms by Staphylococcus aureus, methicillin resistant S. aureus, Escherichia coli, and Candida albicans. Apart from inhibiting the formation of biofilm, pomegranate extract disrupted pre-formed biofilms and inhibited germ tube formation, a virulence trait, in C. albicans. Characterization of the methanolic extract of pomegranate revealed the presence of ellagic acid (2,3,7,8-tetrahydroxy-chromeno[5,4,3-cde]chromene-5,10-dione) as the major component. Ellagic acid is a bioactive tannin known for its antioxidant, anticancer, and anti-inflammatory properties. Further studies revealed the ability of ellagic acid to inhibit the growth of all species in suspension at higher concentrations (>75 μg ml(-1)) and biofilm formation at lower concentrations (<40 μg ml(-1)) which warrants further investigation of the potential of ellagic acid or peel powders of pomegranate for the treatment of human ailments.
    Biofouling 08/2013; · 3.40 Impact Factor
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    ABSTRACT: Group A Streptococcus (Streptococcus pyogenes) is responsible for a wide array of infections and incidence is high in developing countries like India. Although distribution of emm types of S. pyogenes in India has been described, its association with the virulence genes and ocular isolates is less concentrated. In the present study emm type surveillance as well as its association with toxin gene profile was analyzed. Ocular infected cases such as lacrimal abscess, corneal ulcers, mucocoele showed the presence of 20 S. pyogenes isolates. For noninvasive isolates, we screened 370 pharyngitis cases and 400 asymptomatic school children and recovered 33 pharyngitis and 14 carrier isolates respectively. 14 emm type distributions were observed in ocular isolates, 11 emm types each in pharyngitis and asymptomatic carrier isolates. The two dominant emm types, emm49 and emm63 were accounted for 33% of the total S. pyogenes isolates. Among ocular isolates, slo, smeZ, speB and speG were found in >50% of isolates, in pharyngitis smeZ (48%), speB (45%) and speG (42%) genes were found to be prevalent. Alarmingly, carrier isolates showed more prevalence to virulence genes than the ocular and pharyngitis isolates with speF (79%), speB, speG (64%), slo and sil (64%). Among the three groups, pharyngitis isolates harbored more prtF1 (33%) and prtF2 (94%) than the asymptomatic carriers (28% and 71%) and the ocular isolates (45% and 40%). 450bp size band in prtF1 and 350bp size band in prtF2 showed dominance. Among the three groups tested, the distribution of ermB and mefA was high in pharyngitis isolates (30%) where 10 isolates showed the presence of both genes. None of the isolates showed the presence of ermA and tetO genes. Dendrogram generated based on the virulence and antibiotic resistance gene profiles revealed that except one cluster, all other clusters showed some correlation with ocular, pharyngitis and asymptomatic carrier isolates, irrespective of their emm types.
    Infection, genetics and evolution: journal of molecular epidemiology and evolutionary genetics in infectious diseases 07/2013; · 3.22 Impact Factor
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    ABSTRACT: Quorum sensing (QS) is a process of cell-cell communication mechanism occurs between the bacterial cells through the secretary signal molecules. This QS mechanism has been shown to control over the expression of various genes responsible for the production of virulence factors in several bacterial pathogens. Hence, the present study was intended to evaluate the antipathogenic potential of mangrove trees of the genus Rhizophora against the QS dependent virulence factors production in Pseudomonas aeruginosa PAO1, clinical isolates CI-I (GU447237) and CI-II (GU447238). The methanol extract of Rhizophora apiculata and R. mucronata (1mg/ml) showed significant inhibition against QS dependent virulence factors production such as LasA protease, LasB elastase, total protease, pyocyanin pigment production and biofilm formation in P. aeruginosa PAO1, CI-I and CI-II. This study for the first time, reports the quorum sensing inhibitory (QSI) potential of Rhizophora spp. against P. aeruginosa infections.
    Phytomedicine: international journal of phytotherapy and phytopharmacology 06/2013; · 2.97 Impact Factor
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    ABSTRACT: Quinolone resistance-determining region is known to be the druggability site of the target protein that undergoes frequent mutation and thus renders quinolone resistance. In the present study, ligands were tested for their inhibitory activity against DNA gyrase of Streptococcus pyogenes involved in DNA replication. In silico mutational analysis on modelled gyrase A revealed that GLU85 had the most possible interactions with all the ligands used for the study. The amino acid residue GLU85 had also been predicted with an essential role of maintaining the three-dimensional structure of the protein. When introduced with a mutation (GLU 85 LYS) on this particular residue, it had readily denatured the whole α-helix (from 80 to 90 amino acids). This was confirmed through the molecular dynamics simulation and revealed that this single mutation can cause many functional and structural changes. Furthermore, LYS85 mutation has altered the original secondary structure of the protein, which in turn led to the steric hindrance during the ligand-receptor interaction. The results based on the G-score revealed that ligands have reduced interaction with the mutant protein. The semisynthetic fluoroquinolone 6d, which is an exception, forms a strong interaction with the mutant protein and was experimentally verified using the antimicrobial test. Hence, the present study unravels the fact that mutation at the drug binding site is the major cause for different level of resistance by the S. pyogenes when exposed against the varying concentrations of the fluoroquinolones. Furthermore, a comparative assessment of quinolone derivative with the older generation fluoroquinolones will be of great impact for S. pyogenes-related infections. Copyright © 2013 John Wiley & Sons, Ltd.
    Journal of Molecular Recognition 06/2013; 26(6):276-85. · 3.01 Impact Factor
  • Balu Jancy Kalpana, Shunmugiah Karutha Pandian
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    ABSTRACT: A halotolerant α-amylase having the ability of digesting the insoluble raw starches was characterized from Bacillus subtilis S8-18, a marine sediment isolate from Palk Bay region. The electrophoresis techniques unveiled that the α-amylase was indeed a monomer with a molecular weight of 57 kDa. The optimum temperature and pH for the enzyme activity were 60 °C and 6.0 respectively. The enzyme was highly stable for 24 h over a wide range of pH from 4.0 to 12.0 by showing 84-94% activity. Interestingly, by retaining 72% activity even after 24 h, the enzyme also showed tolerance towards 28% NaCl. The α-amylase retained a minimum of 93% residual activity in 1 mM concentration for the selected divalent metal ions. The enzyme was found to be chelator resistant as it remained unaffected by 1 mM of EDTA and exhibited 96% activity even at 5 mM concentration. Furthermore, though 1% SDS caused remarkable reduction (68%) in amylase activity, the enzyme showed tolerance towards other detergents (1% of Triton-X and Tween 80) with 85% activity. Additionally, the α-amylase enzyme is capable of hydrolyzing the insoluble raw starch substrates which was evident from the scanning electron microscopic (SEM) and spectrophotometric analyses.
    Journal of Basic Microbiology 05/2013; 54(8). · 1.20 Impact Factor
  • Balu Jancy Kalpana, Muthukrishnan Sindhulakshmi, Shunmugiah Karutha Pandian
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    ABSTRACT: The present study is aimed at developing an economical medium for the production of α-amylase from Bacillus subtilis S8-18, a marine sediment isolate from Palk Bay, with various agricultural by-products which are cheap and rich in starch. These products included wheat bran, wheat husk, rice bran, rice husk and potato peel and used to replace soluble starch present in the LB broth (synthetic medium). The rice husk was found to be the best to influence enzyme production significantly (61186 IU mL(-1) ) when compared to the yield of 30026 IU mL(-1) obtained by commercial starch. Hence, LB broth containing rice husk was termed as economical medium. Besides, the effect of various nutritional and physiological factors on enzyme production was also investigated. Furthermore, the desizing efficiency of α-amylases produced by synthetic and economical medium was evaluated through various assays like reducing sugar estimation, weight loss assay, drop absorbency assay, SEM and FTIR analyses. In addition, a commercial α-amylase from B. subtilis was also used in desizing analyses for comparative purpose. It revealed that the α-amylase from economical medium was highly effective in desizing the cotton fabrics as that of the commercial enzyme and much superior to the enzyme produced through synthetic medium. This article is protected by copyright. All rights reserved.
    Biotechnology and Applied Biochemistry 05/2013; 61(2). · 1.35 Impact Factor
  • Kannan Balaji, Ramalingam Thenmozhi, Shunmugiah Karutha Pandian
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    ABSTRACT: Background & objectives: Subinhibitory concentrations (sub-MICs) of antibiotics, although not able to kill bacteria, but influence bacterial virulence significantly. Fluoroquinolones (FQs) which are used against other bacterial pathogens creates resistance in non-targeted Streptococcus pyogenes. This study was undertaken to characterize the effect of sub-MICs of FQs on S. pyogenes biofilm formation. Methods: Biofilm forming six M serotypes M56, st38, M89, M65, M100 and M74 of S. pyogenes clinical isolates were challenged against four FQs namely, ciprofloxacin, ofloxacin, levofloxacin and norfloxacin. The antibiofilm potential of these FQs was analysed at their subinhibitory concentrations (1/2 to 1/64 MIC) using biofilm assay, XTT reduction assay, scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). Results: Among the four FQs tested, ofloxacin and levofloxacin at 1/2 MIC showed the maximum inhibition (92%) of biofilm formation against M56 and M74 serotypes. FQs effectively interfered in the microcolony formation of S. pyogenes isolates at 1/2 to 1/8 sub-MICs. Inhibition of biofilm formation was greatly reduced beyond 1/16 MICs and allowed biofilm formation. XTT reduction assay revealed the increase in metabolic activity of S. pyogenes biofilm against the decrease in FQs concentration. SEM and CLSM validated the potential of sub-MICs of FQs against the six S. pyogenes. Interpretation & conclusions: Our results showed that the inhibitory effect all four FQs on S. pyogenes biofilm formation was concentration dependent. FQs at proper dosage can be effective against S. pyogenes and lower concentrations may allow the bacteria to form barriers against the antibiotic in the form of biofilm.
    The Indian Journal of Medical Research 05/2013; 137(5):963-71. · 1.66 Impact Factor
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    ABSTRACT: Solanum trilobatum L. is an Indian medicinal plant containing rich amount of steroidal glyco-alkoloids that can be used as precursor for commercial steroid production. Two efficient marker systems such as Random Amplified Polymorphic DNA (RAPD) and Inter Simple Sequence Repeats (ISSR) were used for the first time to assess the genetic diversity across 14 S. trilobatum accessions obtained from five South Indian states. Twenty out of 60 RAPD primers generated 189 distinct bands of which 160 were polymorphic with an average of 8 polymorphic bands per primer. A maximum of up to 15 fragments were amplified with an average of 9.45 bands per primer and the amplicons varied in size between 100 and 3,000 bp. The percentage of polymorphism ranged from 55.5 to 100, with an average of 84.6. ISSR profiling using 7 out of 20 primers amplified 83 bands and the number of amplified fragments varied from 2 to 16 with a size range of 200–1,800 bp. Totally 72 polymorphic bands were obtained using 7 ISSR primers at an average of 10.28 polymorphic bands per primer. Polymorphism percentage varied from 50 to 100 among the selected accessions resulting in an average percentage of polymorphism of 86.7. The PIC values ranged from 0.49 to 0.93 for RAPD and 0.16 to 0.90 for ISSR primers. The study pointed out that ISSR markers were more efficient than RAPD markers in evaluating the degree of genetic variation in S. trilobatum. The UPGMA cluster analysis grouped all Tamil Nadu accessions in one cluster and other state accessions in another cluster. The Principal component analysis also substantiates this clustering pattern. Thus the phylogenetic relationship and a high genetic variation revealed in the present study could provide a baseline data for conservation and improvement of this plant in future. Also the molecular markers identified in this study will be helpful in authentication of this species to prevent adulteration in herbal medicine.
    Genetic Resources and Crop Evolution 03/2013; 60(3). · 1.48 Impact Factor