I. S. Bright Singh

Bio: I. S. Bright Singh is an academic researcher from Cochin University of Science and Technology. The author has contributed to research in topics: Penaeus monodon & Shrimp. The author has an hindex of 24, co-authored 120 publications receiving 1876 citations.

ISSN 2249-9687 Original Article Isolation, Purification and Characterization of Antimicrobial Peptides from Indian Ruff, Psenopsis cyanea (Alcock, 1890).

Abstract: Antimicrobial peptides are evolutionarily ancient, effector molecules of innate immune defense and are ubiquitously found in all kingdoms from bacteria to mammals, including fungi and plants. The present study deals with the isolation of antimicrobial peptides from marine bathydemersal fish Indian ruff, Psenopsis cyanea belonging to the family Centrolophidae and its in vitro antimicrobial activity. Peptides were extracted from the sample by modified acetic acidacetone precipitation method.The crude peptide extract exhibited antimicrobial activity against bacteria and fungi. Prepurification was done by solid phase extraction using Sep-Pak ® C- 18 cartridges and 40% Sep-Pak eluted fractions were subjected to FPLC using UNO TM QI column. Four fractions eluted on the basis of retention time were collected and further examined for antimicrobial activity by disc diffusion assay. FPLC fraction P40-1 was found to be active only against A. hydrophila and P40-2 was found to be active against Gram-positive and Gram-negative strains including S. aureus, B. cereus, and A. hydrophila. Whereas, P40-3 only showed inhibitory effect on A. hydrophila. Among the various fractions, P40-4 was the only fraction inhibited the growth of V. fluvialis. Active fractions were subjected to protein quantification. This is the first report on isolation and purification of AMPs from Psenopsis cyanea. © 2015 Universal Research Publications. All rights reserved

Molecular characterization and phylogenetic analysis of an anti-lipopolysaccharide factor from the crucifix crab, Charybdis feriatus

Abstract: m Abstract: Antimicrobial peptides are small cationic, gene-encoded, amphipathic, host defense peptides with a ubiquitous distribution in all living kingdoms. They are ,10 kDa in size, with 15-100 amino acids having a net positive charge of +2 to +9. Anti-lipopolysaccharide factor (ALF) is a cationic antimicrobial peptide which constitutes one of the key effector molecules in the innate immune system of crustaceans, and is capable of binding and neutralizing lipopoly- saccharides. In the present study, an ALF homolog (Charybdis feriatus (Cf)-ALF1)-encoding cDNA sequence from the hemocytes of the crucifix crab, C. feriatus, was cloned, identified, and characterized. The deduced peptide of Cf-ALF1 encoded for a 123 amino acid peptide with a 97 residue mature peptide (11.16 kDa) that had a net charge of +10. Two conserved cysteine residues and a putative lipopolysaccharide binding domain were observed in the Cf-ALF1 mature peptide. BLAST analysis of Cf-ALF1 nucleotides showed a 99% similarity to Scylla serrata. The spatial structure of Cf-ALF1 was composed of three α-helices packed against a four-strand β-sheet. Two of these helices were linked by a disulfide bond to form an amphipathic loop similar to the structure of anti-lipopolysaccharide factor isoform 3 from Penaeus monodon (ALF-Pm3). All these features suggest that Cf-ALF1 could play a significant role in the innate immune defense mechanism of C. feriatus.

Molecular and Functional Characterization of an Anti-lipopolysaccharide Factor Mm-ALF from Speckled Shrimp Metapenaeus monoceros

Abstract: Anti-lipopolysaccharide factors (ALFs) are antimicrobial peptides of approximately 100 amino acid residues with a broad spectrum of antimicrobial activity It is an amphipathic peptide with an N-terminal hydrophobic region and a lipopolysaccharide binding domain (LBD) In the present study, we report an isoform of the anti-lipopolysaccharide factor (Mm-ALF) from the speckled shrimp, Metapenaeus monoceros A 359 bp cDNA encoded 119 amino acids, and the sequence showed 9916% similarity to ALF from the shrimp Fenneropenaeus indicus The mature peptide of 94 amino acids has a net charge of +8, molecular weight 1062 kDa, and pI 1011 The mature peptide Mm-ALF was recombinantly expressed in E coli Rosetta-gami cells, and the peptide was isolated and purified The rMm-ALF exhibited notable antibacterial activity against Gram-positive (Staphylococcus aureus and Bacillus cereus) and Gram-negative (Escherichia coli, Edwardsiella tarda, Aeromonas hydrophila, Pseudomonas aeruginosa, Vibrio parahaemolyticus, Vibrio harveyi, Vibrio alginolyticus, Vibrio proteolyticus, Vibrio cholerae and Vibrio fluvialis) bacteria

Purification and characterization of highly active LasB protease from Pseudomonas aeruginosa MCCB 123

Abstract: Proteolytic enzymes constitute one of the most important groups of industrial enzymes and have received worldwide attention due to their wide range of biotechnological applications and their sustainability in reducing environmental degradation. In this study, a LasB protease with dual substrate specificities was purified from Pseudomonas aeruginosa MCCB 123 with 6.85 fold increase in protease specific activity and 9.05 fold increase in elastase-specific activity. The enzyme revealed the presence of a 33 kDa protein. Optimum temperature and pH of protease activity were 60°C and 9.0, and that of elastase activity were 40°C and 8.0, respectively. Based on the cytotoxicity assay on Hep2 cells, the enzyme has an IC50 (inhibitory concentration) value of 47.28 μg mL . According to the deduced amino acid sequence, the MCCB 123 LasB protease is a zinc dependent metalloendopeptidase belonging to M4 neutral protease GluZincin superfamily. Appreciable protease activity of the enzyme at high temperature and alkaline pH proposes its potential application in detergency. MCCB 123 LasB protease is 3.96 fold more active when compared to commercial protease Savinase suggesting its industrial relevance.

The Design and Analysis of Experiments

Abstract: THE DESIGN AND ANALYSIS OF EXPERIMENTS. By Oscar Kempthorne. New York, John Wiley and Sons, Inc., 1952. 631 pp. $8.50. This book by a teacher of statistics (as well as a consultant for \"experimenters\") is a comprehensive study of the philosophical background for the statistical design of experiment. It is necessary to have some facility with algebraic notation and manipulation to be able to use the volume intelligently. The problems are presented from the theoretical point of view, without such practical examples as would be helpful for those not acquainted with mathematics. The mathematical justification for the techniques is given. As a somewhat advanced treatment of the design and analysis of experiments, this volume will be interesting and helpful for many who approach statistics theoretically as well as practically. With emphasis on the \"why,\" and with description given broadly, the author relates the subject matter to the general theory of statistics and to the general problem of experimental inference. MARGARET J. ROBERTSON