Lipase

Introduction Enzymes Etymology and historical aspects of Industrial enzymes Industrial enzymes: development of bio-industrial sector Classification of Enzymes Industrially important enzymes Kinetic model of lipolysis, structure of lipase, stability and cofactors Classification of bacterial lipolytic enzymes Application of Lipases Detergent industry Food processing, flavor development and improving quality Bakery industry Fat and oleochemical industry Textile industry Cosmetic industry Pulp and paper industry Oil biodegradation Production of biodegradable polymers Diagnostic tool Degreasing of leather Waste/effluent/sewage treatment Production of biodiesel Tea processing Biosensors Environmental Management Major obstacles and future prospect of microbial lipases Metagenomics and Unculturable Bacteria Microbial communities Soil as a microbial habitat Unculturable microorganisms Culturable microorganisms Microbial species for beneficial products and processes Metagenomics Soil metagenomics for desirable genes Soil metagenomics: tool for novel compounds Sequence based metagenomics Function driven metagenomics Metagenomic library construction and its analysis Approaches to metagenomics Accessing Metagenomics Methods Expression vector/host system Functional screening of metagenomic libraries Metagenomics for Lipase Collection of environmental samples Extraction, purification and quantification of soil metagenomic DNA Isolation and purification of mgDNA from gram+ and gram- bacteria Polymerase chain reaction Restriction digestion by Eco RI, Hind III and Bam HI Metagenomic DNA library construction Sequence based metagenomic approach Construction of 16S rDNA gene libraries Preparation of E. coli competent cells and transformation Analysis of transformation efficiency using non-recombinant plasmid Screening of recombinants by alpha-complementation Purification of PCR products Transformant confirmation by colony PCR using universal primers Isolation and restriction of recombinant/non-recombinant plasmid DNAs Analysis of 16S rDNA gene Phylogenetic analysis Genbank submission and accession numbers Phylogeny Functional Approach for Metagenomic Library Construction Restriction digestion of mgDNA Transformation of E. coli BL21 competent cells Functional screening of metagenomic libraries for the lipase gene Isolation and restriction of recombinant plasmid Sequencing of cloned DNA Sequence analysis and phylogenetic tree construction Analysis of the cloned lipase gene Phylogeny of recombinant protein Amino acid sequence of recombinant protein Multiple sequence alignment Storage of recombinant plasmid Overexpression of Recombinant Protein Purification of expressed protein Determination of active lipase using zymographic study Determination of specific activity, % yield and protein fold purification Homology model and validation for protein structure prediction Biochemical Characterization of Purified Lipase Dose dependent enzyme activity Substrate specificity and effect of substrate concentration Enzyme Kinetics Effect temperature and pH Effect of surfactants Effect of other factors Genomic Study of Culture Dependent Bacteria Isolation and screening for lipase producing culturable bacteria Pure culture of lipase producing bacterial isolates Inoculum preparation Zone of hydrolysis Growth kinetics and lipase production Thermal stability Evaluation of crude enzyme for detergent formulation Stability of culture supernatant under storage Taxonomic identification of lipase producing bacteria Biochemical characterization Extracellular enzyme activity Spectrophotometric growth determination Genomic Study of Culturable Bacteria Genomic DNA extraction of lipase producing culturable bacteria Molecular genetic assessment of lipase producing bacteria Phylogenetic analysis Optimization of culture condition for lipase production Effect of carbon and nitrogen sources for the growth of culturable bacterial Time course with lipase production Effect of other factors Microbial Assay of Culture Supernatant Containing Crude Lipase Determination of antibacterial activity Determination of antifungal activity Critical Observations