Production, purification, genetic characterization and application studies of tannase enzyme from marine fungus Aspergillus awamori

Marine fungi remain totally unexplored as a source of industrial enzyme and prospective applications. Further tannase production by a marine organism has so far not been established. The primary objective of this study included the evaluation of the potential of Aspergillus awamori isolated from sea water as part of an earlier study and available in the culture collection of the Microbial technology laboratory for tannase production through different fermentation methods, optimization of bioprocess variables by statistical methods, purification and characterization of the enzyme, genetic study, and assessment of its potential applications.

α-Galactosidase from Streptomyces griseoloalbus: an enzyme with versatile applications

The present study is focused on the production, purification and characterization of multiple thermostable α-galactosidases from a novel actinomycete strain Streptomyces griseoloalbus. The Chapter I of the thesis covers the wide literature regarding α-galactosidases from various sources and their potential applications. The Chapter 11 deals with the isolation of α-galactosidase- producing actinomycetes and selection of the best strain. The Chapters III and IV describe the optimization of α-galactosidase production under submerged fermentation and solid-state fermentation respectively. The Chapter V describes the purification and characterization of multiple α-galactosidases and also the obvious existence of a novel galactose-tolerant enzyme. The Chapter VI illustrates the potential applications of α-galactosidases from S. griseoloalbus followed by the Chapter VII summarizing and concluding the results of the present investigation.

Optimization of Lignin Peroxidase, Manganese Peroxidase, and Lac Production from Ganoderma lucidum Under Solid State Fermentation of Pineapple Leaf

This study was undertaken to isolate ligninase-producing white-rot fungi for use in the extraction of fibre from pineapple leaf agriwaste. Fifteen fungal strains were isolated from dead tree trunks and leaf litter. Ligninolytic enzymes (lignin peroxidase (LiP), manganese peroxidase (MnP), and laccase (Lac)), were produced by solid-state fermentation (SSF) using pineapple leaves as the substrate. Of the isolated strains, the one showing maximum production of ligninolytic enzymes was identified to be Ganoderma lucidum by 18S ribotyping. Single parameter optimization and response surface methodology of different process variables were carried out for enzyme production. Incubation period, agitation, and Tween-80 were identified to be the most significant variables through Plackett-Burman design. These variables were further optimized by Box-Behnken design. The overall maximum yield of ligninolytic enzymes was achieved by experimental analysis under these optimal conditions. Quantitative lignin analysis of pineapple leaves by Klason lignin method showed significant degradation of lignin by Ganoderma lucidum under SSF

BIOPROCESS OPTIMIZATION AND CHARACTERIZATION OF PHYTASE FROM AN ENVIRONMENTAL ISOLATE BACILLUS MCCB 242

A study was undertaken to isolate phytase producers from environment and to segregate the most highly efficient phytase producer and to develop a bioprocess technology for commercial application. During this process, a potential phytase producer Bacillus MCCB 242 was isolated and characterized phenotypically and genotypically. Subsequently, phytase production was optimized, the enzyme purified and characterized and an appropriate downstream process also could be standardized.Precisely, through this work an environmental isolate Bacillus MCCB 242 could be brought out as phytase producer for commercial application. The enzyme production could be optimized and characterized, and an appropriate downstream process standardized. Cytotoxicity studies revealed the enzyme safe for feed application, especially in fish.

Optimization of pH and Direct Imaging Conditions of Complexed Methylene Blue Sensitized Poly(vinyl chloride) Films

Significant results of our experimental investigations on the dependence of pH on real time transmission characteristics on recording media fabricated by doping PVC with complexed methylene blue are presented. The optimum pH value for faster bleaching was found to be 4×5. In typical applications, the illumination from one side, normal to the surface of this material, initiates a chemical sequence that records the incident light pattern in the polymer. Thus direct imaging can be successfully done on this sample. The recorded letters were very legible with good contrast and no scattering centres. Diffraction efficiency measurements were also carried out on this material.

Optimization of pH and Direct Imaging Conditions of Complexed Methylene Blue Sensitized Poly(vinyl chloride) Films

Significant results of our experimental investigations on the dependence of pH on real time transmission characteristics on recording media fabricated by doping PVC with complexed methylene blue are presented. The optimum pH value for faster bleaching was found to be 4×5. In typical applications, the illumination from one side, normal to the surface of this material, initiates a chemical sequence that records the incident light pattern in the polymer. Thus direct imaging can be successfully done on this sample. The recorded letters were very legible with good contrast and no scattering centres. Diffraction efficiency measurements were also carried out on this material.

Optimization of pH and direct imaging conditions of complexed methylene blue sensitized poly(vinyl chloride) films

Significant results of our experimental investigations on the dependence of pH on real time transmission characteristics on recording media fabricated by doping PVC with complexed methylene blue are presented. The optimum pH value for faster bleaching was found to be 4 . 5. In typical applications, the illumination from one side, normal to the surface of this material, initiates a chemical sequence that records the incident light pattern in the polymer. Thus direct imaging can be successfully done on this sample. The recorded letters were very legible with good contrast and no scattering centres. Diffraction efficiency measurements were also carried out on this material.

Alkaline protease production by marine fungus engyodontium BTMFS 10

The thesis entitled “Alkaline Protease Production by Marine Fungus Engyodontium BTMFS 10”.Proteases are the single class of enzymes, which occupy a pivotal position with respect to their application in both physiological and commercial filed. Protease in the industrial market is expected to increase further in the coming year. The current trend is to use microbial enzymes since they provide a greater diversity of catalytic activities and can be produced more economically. Main objective of theses studies are the optimization of various physicochemical factors in the solid state fermentation for the production of alkaline protease enzyme, characterization of the enzyme, evaluation of the enzyme for various industrial application. The result obtained the during the course of theses study indicate the scope for the utilization of this study Marine Fungus E. Album for extra cellular protease production employing solid state fermentation

Optimization of carbon and nitrogen sources and growth factors for the production of an aquaculture probiotic (Pseudomonas MCCB 103) using response surface methodology

Aim: To develop a new medium for enhanced production of biomass of an aquaculture probiotic Pseudomonas MCCB 103 and its antagonistic phenazine compound, pyocyanin. Methods and Results: Carbon and nitrogen sources and growth factors, such as amino acids and vitamins, were screened initially in a mineral medium for the biomass and antagonistic compound of Pseudomonas MCCB 103. The selected ingredients were further optimized using a full-factorial central composite design of the response surface methodology. The medium optimized as per the model for biomass contained mannitol (20 g l)1), glycerol (20 g l)1), sodium chloride (5 g l)1), urea (3Æ3 g l)1) and mineral salts solution (20 ml l)1), and the one optimized for the antagonistic compound contained mannitol (2 g l)1), glycerol (20 g l)1), sodium chloride (5Æ1 g l)1), urea (3Æ6 g l)1) and mineral salts solution (20 ml l)1). Subsequently, the model was validated experimentally with a biomass increase by 19% and fivefold increase of the antagonistic compound. Conclusion: Significant increase in the biomass and antagonistic compound production could be obtained in the new media. Significance and Impact of the Study: Media formulation and optimization are the primary steps involved in bioprocess technology, an attempt not made so far in the production of aquaculture probiotics.

Effect of Dietary Proteins on Growth, Food Conversion and Digestive Enzyme Activity of Juvenile Macrobrachium Rosenbergii

School of Industrial Fisheries, Cochin University of Science and Technology

Optimization of Preparation Techniques and Dielectric Study of Polyanilines in the Microwave & High Frequency Field

The thesis deals with the preparation and dielectric characterization of Poly aniline and its analogues in ISM band frequency of 2-4 GHz that includes part of the microwave region (300 MHz to 300 GHz) of the electromagnetic spectrum and an initial dielectric study in the high frequency [O.05MHz-13 MHz]. PolyaniIine has been synthesized by an in situ doping reaction under different temperature and in the presence of inorganic dopants such as HCl H2S04, HN03, HCl04 and organic dopants such as camphorsulphonic acid [CSA], toluenesulphonic acid {TSA) and naphthalenesulphonic acid [NSA]. The variation in dielectric properties with change in reaction temperature, dopants and frequency has been studied. The effect of codopants and microemulsions on the dielectric properties has also been studied in the ISM band. The ISM band of frequencies (2-4 GHz) is of great utility in Industrial, Scientific and Medical (ISM) applications. Microwave heating is a very efficient method of heating dielectric materials and is extensively used in industrial as well as household heating applications.

Glucoamylase immobilized on montmorillonite: influence of nature of binding on surface properties of clay-support and activity of enzyme

Glucoamylase was immobilized on acid activated montmorillonite clay via two different procedures namely adsorption and covalent binding. The immobilized enzymes were characterized by XRD, NMR and N2 adsorption measurements and the activity of immobilized glucoamylase for starch hydrolysis was determined in a batch reactor. XRD shows intercalation of enzyme into the clay matrix during both immobilization procedures. Intercalation occurs via the side chains of the amino acid residues, the entire polypeptide backbone being situated at the periphery of the clay matrix. 27Al NMR studies revealed the different nature of interaction of enzyme with the support for both immobilization techniques. N2 adsorption measurements indicated a sharp drop in surface area and pore volume for the covalently bound glucoamylase that suggested severe pore blockage. Activity studies were performed in a batch reactor. The adsorbed and covalently bound glucoamylase retained 49% and 66% activity of the free enzyme respectively. They showed enhanced pH and thermal stabilities. The immobilized enzymes also followed Michaelis–Menten kinetics. Km was greater than the free enzyme that was attributed to an effect of immobilization. The immobilized preparations demonstrated increased reusability as well as storage stability.

Process Optimization for Mass Production of Marine Yeast Candida sp. S 27 and its Nutritional Characterization

The main source of protein for human and animal consumption is from the agricultural sector, where the production is vulnerable to diseases, fluctuations in climatic conditions and deteriorating hydrological conditions due to water pollution. Therefore Single Cell Protein (SCP) production has evolved as an excellent alternative. Among all sources of microbial protein, yeast has attained global acceptability and has been preferred for SCP production. The screening and evaluation of nutritional and other culture variables of microorganisms are very important in the development of a bioprocess for SCP production. The application of statistical experimental design in bioprocess development can result in improved product yields, reduced process variability, closer confirmation of the output response to target requirements and reduced development time and overall cost.The present work was undertaken to develop a bioprocess technology for the mass production of a marine yeast, Candida sp.S27. Yeasts isolated from the offshore waters of the South west coast of India and maintained in the Microbiology Laboratory were subjected to various tests for the selection of a potent strain for biomass production. The selected marine yeast was identified based on ITS sequencing. Biochemical/nutritional characterization of Candida sp.S27 was carried out. Using Response Surface Methodology (RSM) the process parameters (pH, temperature and salinity) were optimized. For mass production of yeast biomass, a chemically defined medium (Barnett and Ingram, 1955) and a crude medium (Molasses-Yeast extract) were optimized using RSM. Scale up of biomass production was done in a Bench top Fermenter using these two optimized media. Comparative efficacy of the defined and crude media were estimated besides nutritional evaluation of the biomass developed using these two optimized media.