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Nagarajan Kayalvizhi,Paramasamy Gunasekaran 한국생물공학회 2010 Biotechnology and Bioprocess Engineering Vol.15 No.2
A bacterial strain Bacillus licheniformis MKU3,isolated from slaughterhouse sediments showed a strong antimicrobial activity. The antimicrobial substance produced by this strain was found to be a protein that inhibited a broad range of bacterial strains, such as Bacillus sp.,Staphylococcus sp., Streptococcus sp., and Listeria monocytogenes. The antimicrobial peptide was purified to homogeneity by cut off membrane filtration followed by gel filtration chromatography. The purified protein with low molecular mass (< 8 kDa) was resolved as single band on Tricine SDS-PAGE. This protein was stable at 100oC for 10 min, but lost its activity at 121oC in 15 min. It was resistant to the proteolytic action of trypsin, proteinase K,and pronase E and stable within a wide range of pH (3.0~11.0). This protein exhibited lytic activity on selected indicator strain Kurthia gibsonii GCS6.
Ganesan Arun,Muthukumarasamy Eyini,Paramasamy Gunasekaran 한국생물공학회 2014 Biotechnology and Bioprocess Engineering Vol.19 No.6
Silver nanoparticles, which are small metalliccolloidal particles, ranging 1 ~ 100 nm in size, have severalnano biotechnological applications in medicine, manufacturingand engineering industries. Fungus-mediated synthesis ofsilver nanoparticles is an ecofriendly, green process. Further,extracellular enzymes and proteins elaborated by fungi areinvolved in the synthesis of the silver nanoparticle, whichmakes the downstream processing relatively simpler. In thepresent investigation, Schizophyllum commune, a mushroomfungus, was tested for its ability to synthesize extracellularas well as intracellular silver nanoparticles. When thefungus was challenged with 1 mM silver nitrate, a changein colour of the broth and the mycelium was observed,indicative of extracellular and intracellular synthesis ofsilver nanoparticles. The presence of silver nanoparticleswas confirmed by studying its Surface Plasmon Resonanceabsorption band in the visible wavelength. FTIR spectrumanalysis of the silver nanoparticles indicated the presenceof biomolecules in association with the reduction of silverions. Scanning Electron Microscopic analysis of the silvernanoparticles revealed the nanorange dimensions of boththe extracellular and the intracellular silver nanoparticles. Analysis of biological activities of the silver nanoparticlesdisclosed their significant antibacterial activity againstEscherichia coli, Bacillus subtilis, Klebsiella pneumoniaeand Pseudomonas fluorescens. Additionally, the silvernanoparticles inhibited the growth of the dermatophyticfungal pathogens viz. Trichophyton simii, Trichophytonmentagrophytes and Trichophyton rubrum significantly. Anticancer activity of silver nanoparticles, assayed throughMTT cytotoxicity assay, uncovered that 27.2 and 64%mortality could be obtained in Human Epidermoid LarynxCarcinoma (HEP -2) cell lines at concentrations between10 and 100 μg/mL, respectively. The results obtained indicatethat Schizophyllum commune is capable of synthesizingsilver nanoparticles in shaken broth cultures (120 rpm) at25 ± 2°C and pH 7.
Repeated Random Mutagenesis of α-Amylase from Bacillus Licheniformis for Improved pH Performance
( Ramachandran Priyadharshini ),( Shankar Manoharan ),( Devaraj Hemalatha ),( Paramasamy Gunasekaran ) 한국미생물 · 생명공학회 2010 Journal of microbiology and biotechnology Vol.20 No.12
The α-amylases activity was improved by random mutagenesis and screening. A region comprising residues from the position 34-281 was randomly mutated in B. licheniformis α-amylase (AmyL), and the library with mutations ranging from low, medium, and high frequencies was generated. The library was screened using an effective liquid-phase screening method to isolate mutants with an altered pH profile. The sequencing of improved variants indicated 2-5 amino acid changes. Among them, mutant TP8H5 showed an altered pH profile as compared with that of wild type. The sequencing of variant TP8H5 indicated 2 amino acid changes, Ile157Ser and Trp193Arg, which were located in the solvent accessible flexible loop region in domain B.
( Rameshkumar ),( Neelamegam ),( Niraikulam Ayyadurai ),( Nagarajan Kayalvizhi ),( Paramasamy Gunasekaran ) 한국미생물 · 생명공학회 2012 Journal of microbiology and biotechnology Vol.22 No.1
The genetic diversity of plant growth-promoting rhizobacterial (PGPR) fluorescent pseudomonads associated with the sugarcane (Saccharum officinarum L.) rhizosphere was analyzed. Selected isolates were screened for plant growthpromoting properties including production of indole acetic acid, phosphate solubilization, denitrification ability, and production of antifungal metabolites. Furthermore, 16S rDNA sequence analysis was performed to identify and differentiate these isolates. Based on 16S rDNA sequence similarity, the isolates were designated as Pseudomonas plecoglossicida, P. fluorescens, P. libaniensis, and P. aeruginosa. Differentiation of isolates belonging to the same group was achieved through different genomic DNA fingerprinting techniques, including randomly amplified polymorphic DNA (RAPD), amplified ribosomal DNA restriction analysis (ARDRA), repetitive extragenic palindromic (REP), enterobacterial repetitive intergenic consensus (ERIC), and bacterial repetitive BOX elements (BOX) analyses. The genetic diversity observed among the isolates and rep-PCR-generated fingerprinting patterns revealed that PGPR fluorescent pseudomonads are associated with the rhizosphere of sugarcane and that P. plecoglossicida is a dominant species. The knowledge obtained herein regarding the genetic and functional diversity of fluorescent pseudomonads associated with the sugarcane rhizosphere is useful for understanding their ecological role and potential utilization in sustainable agriculture.