Development of Bioreactors for Nitrifying Water In Closed System Hatcheries Of Penaeid And Non-Penaeid Prawns

In the present study the development of bioreactors for nitrifying water in closed system hatcheries of penaeid and non-penaeid prawns. This work is an attempt in this direction to cater to the needs of aquaculture industry for treatment and remediation of ammonia and nitrate in penaeid and non-penaeid hatcheries, by developing nitrifying bacteria allochthonous to the particular environment under consideration, and immobilizing them on an appropriately designed support materials configured as reactors. Ammonia toxicity is the major limiting factors in penaeid and non-penaeid hatchery systems causing lethal and sublethal effects on larvae depending on the pH values. Pressing need of the aquaculture industry to have a user friendly and economically viable technology for the removal of ammonia, which can be easily integrated to the existing hatchery designs without any major changes or modifications. Only option available now is to have biological filters through which water can be circulated for the oxidation of ammonia to nitrate through nitrite by a group of chemolithotrophs known as nitrifying bacteria. Two types of bioreactors have been designed and developed. The first category named as in situ stringed bed suspended bioreactor(SBSBR) was designed for use in the larval rearing tanks to remove ammonia and nitrite during larval rearing on a continuous basis, and the other to be used for nitrifying freshly collected seawater and spent water named as ex situ packed bed bioreactior(PBBR). On employing the two reactors together , both penaeid and non-penaeid larval rearing systems can be made a closed recirculating system at least for a season. A survey of literature revealed that the in situ stringed bed suspended reactor developed here is unique in its design, fabrication and mode of application.

Polyphenolic Compounds Liberated during Coir Retting in a Bio-reactor: Separation, Characterisation and Possible Applications

School of environmental studies, Cochin University of Science and Technology

Fixed bed reactor performance of invertase immobilized on montmorillonite

Invertase was immobilized on acid activated montmorillonite via two independent procedures, adsorption and covalent binding. The immobilized enzymes were characterized by XRD, NMR and N2 adsorption measurements and their activity was tested in a fixed bed reactor. XRD revealed that the enzyme was situated on the periphery of the clay and the side chains of different amino acid residues were involved in intercalation with the clay matrix. NMR demonstrated that tetrahedral Al was linked to the enzyme during adsorption and the octahedral Al was involved during covalent binding. Secondary interaction of the enzyme with Al was also observed. N2 adsorption studies showed that covalent binding of enzymes caused pore blockage since the highly polymeric species were located at the pore entrance. The fixed bed reactor proved to be efficient for the immobilized invertase. The optimum pH and pH stability improved upon immobilization. The kinetic parameters calculated also showed an enhanced efficiency of the immobilized systems. They could be used continuously for long period. Covalently bound invertase demonstrated greater operational stability.

Glucoamylase immobilized on montmorillonite: Synthesis, characterization and starch hydrolysis activity in a fixed bed reactor

Glucoamylase from Aspergillus Niger was immobilized on montmorillonite clay (K-10) by two procedures, adsorption and covalent binding. The immobilized enzymes were characterized using XRD, surface area measurements and 27Al MAS NMR and the activity of the immobilized enzymes for starch hydrolysis was tested in a fixed bed reactor (FBR). XRD shows that enzyme intercalates into the inter-lamellar space of the clay matrix with a layer expansion up to 2.25 nm. Covalently bound glucoamylase demonstrates a sharp decrease in surface area and pore volume that suggests binding of the enzyme at the pore entrance. NMR studies reveal the involvement of octahedral and tetrahedral Al during immobilization. The performance characteristics in FBR were evaluated. Effectiveness factor (η) for FBR is greater than unity demonstrating that activity of enzyme is more than that of the free enzyme. The Michaelis constant (Km) for covalently bound glucoamylase was lower than that for free enzyme, i.e., the affinity for substrate improves upon immobilization. This shows that diffusional effects are completely eliminated in the FBR. Both immobilized systems showed almost 100% initial activity after 96 h of continuous operation. Covalent binding demonstrated better operational stability.

Anaerobic Reactor Development for Complex Organic Wastewater

Faculty of Engineering. Cochin University of Science and Technology

Nitrifying Bioreactors Integrated into Shrimp and Prawn Hatchery Systems: Molecular Characterization of the Nitrifying Consortia,Reactor Kinetics, Modeling and Validation

National Centre for Aquatic Animal Health, Cochin University of Science and Technology

“Studies on extracellular enzyme production during growth of Pleurotus sp. on lignocellulosic agriwaste and the utilization of spent mushroom substrate”

Commercially, Pleurotus spp. of mushroom are cultivated in bags. After mushroom cultivation, spent substrate remains as residual material. Proper recycling of spent substrate is beneficial for our economy. Spent substrate can be utilized for various other value added purposes through the proper knowledge of its components. Composition of various components depends on the activity of extracellular enzymes in the spent substrate. The present study was conducted to know the enzyme profile of some major extracellular enzymes - cellulase, hemicellulase (xylanase), pectinase and ligninase (lignin peroxidase and laccase) and to estimate cellulose, hemicellulose, pectin and lignin in the substrate. The use of spent substrate as a source of fibre and ethanol, and in the biodegradation of phenol by Pleurotus spp. was also investigated

Continuous production of L-glutaminase by an immobilized marine Pseudomonas sp BTMS-51 in a packed bed reactor

A marine Pseudomonas sp BTMS-51, immobilized by Ca-alginate gel entrapment was used for the production of extracellular Lglutaminase under repeated batch process and continuous process employing a packed bed reactor (PBR). Immobilized cells could produce an average of 25 U/ml of enzyme over 20 cycles of repeated batch operation and did not show any decline in production upon reuse. The enzyme yield correlated well with the biomass content in the beads. Continuous production of the enzyme in PBR was studied at different substrate concentrations and dilution rates. In general, the volumetric productivity increased with increased dilution rate and substrate concentrations and the substrate conversion efficiency declined. The PBR operated under conditions giving maximal substrate conversion efficiency gave an average yield of 21.07 U/ml and an average productivity of 13.49 U/ml/h. The system could be operated for 120 h without any decline in productivity

Continuous Production of Extracellular L-Glutaminase by Ca-Alginate-Immobilized Marine Beauveria bassiana BTMF S-10 in Packed-Bed Reactor

L-Glutamine amidohydrolase (L-glutaminase, EC 3.5.1.2) is a therapeutically and industrially important enzyme. Because it is a potent antileukemic agent and a flavor-enhancing agent used in the food industry, many researchers have focused their attention on L-glutaminase. In this article, we report the continuous production of extracellular L-glutaminase by the marine fungus Beauveria bassiana BTMF S-10 in a packed-bed reactor. Parameters influencing bead production and performance under batch mode were optimized in the order-support (Na-alginate) concentration, concentration of CaCl2 for bead preparation, curing time of beads, spore inoculum concentration, activation time, initial pH of enzyme production medium, temperature of incubation, and retention time. Parameters optimized under batch mode for L-glutaminase production were incorporated into the continuous production studies. Beads with 12 × 108 spores/g of beads were activated in a solution of 1% glutamine in seawater for 15 h, and the activated beads were packed into a packed-bed reactor. Enzyme production medium (pH 9.0) was pumped through the bed, and the effluent was collected from the top of the column. The effect of flow rate of the medium, substrate concentration, aeration, and bed height on continuous production of L-glutaminase was studied. Production was monitored for 5 h in each case, and the volumetric productivity was calculated. Under the optimized conditions for continuous production, the reactor gave a volumetric productivity of 4.048 U/(mL·h), which indicates that continuous production of the enzyme by Ca-alginate-immobilizedspores is well suited for B. bassiana and results in a higher yield of enzyme within a shorter time. The results indicate the scope of utilizing immobilized B. bassiana for continuous commercial production of L-glutaminase

Ethanol Production From Spent Substrate Of Pleurotus Eous

Spent substrate, the residual material of mushroom cultivation, causes disposal problems for cultivators. Currently the spent substrate of different mushrooms is used mainly for composting. Edible mushrooms of Pleurotus sp. can grow on a wide range of lignocellulosic substrates. In the present study, Pleurotus eous was grown on paddy straw and the spent substrate was used for the production of ethanol. Lignocellulosic biomass cannot be saccharified by enzymes to high yield of ethanol without pretreatment. The root cause for the recalcitrance of lignocellulosic biomass such as paddy straw is the presence of lignin and hemicelluloses on the surface of cellulose. They form a barrier and prevent cellulase from accessing the cellulose in the substrate. In the untreated paddy straw, the amount of hemicelluloses and lignin (in % dry weight) were 20.30 and 20.34 respectively and the total reducing sugar was estimated to be 5.40 mg/g. Extracellular xylanase and ligninases of P. eous could reduce the amount of hemicelluloses and lignin to 16 and 11(% dry weight) respectively, by 21st day of cultivation. Growth of mushroom brought a seven fold increase in the total reducing sugar yield (39.20 mg/g) and six fold increase in the production of ethanol (6.48 g/L) after 48hrs of fermentation, when compared to untreated paddy straw

Biodegradation Of Phenol Using Spent Substrate Of Pleurotus Sp.

Phenol is an aromatic hydrocarbon which exists as a colorless or white solid in its pure state. Over the past several decades, there is growing concern about wide spread contamination of surface and ground water by phenol, due to rapid development of chemical and petrochemical industries. Phenol affects aquatic life even at relatively low concentration (5-25mg/L). Treatment for removal of phenol includes chemical as well as biological processes. Studies show that ligninases such as Lignin Peroxidase and Laccase, produced by Pleurotus sp., can degrade phenol. Spent substrate of Pleurotus mushrooms consists of ligninases. Present work was to investigate the potential of spent substrate of edible mushroom P. ostreatus for biodegradation of phenol. P. ostreatus was cultivated on paddy straw. After harvest, spent substrate was utilized for phenol degradation. According to the enzyme profile of two ligninases present in the spent substrate of P. ostreatus, maximum specific activity for Laccase was observed in 35 day old spent substrate and LiP activity was maximum in 56 day old spent substrate, which together contributed significantly for removal of phenol. Spent substrate of 35th and 56th day were each incubated with phenol sample (1:1w/v) for one day, which resulted in degradation of phenol by 48% and 45% respectively. From these results it appears that, spent substrate of P. ostreatus can be used effectively to remove phenol from industrial effluents