Propyl Gallate Synthesis Using Acidophilic Tannase and Simultaneous Production of Tannase and Gallic Acid by Marine Aspergillus awamori BTMFW032

Marine Aspergillus awamori BTMFW032, recently reported by us, produce acidophilic tannase as extracellular enzyme. Here, we report the application of this enzyme for synthesis of propyl gallate by direct transesterification of tannic acid and in tea cream solubilisation besides the simultaneous production of gallic acid along with tannase under submerged fermentation by this fungus. This acidophilic tannase enabled synthesis of propyl gallate by direct transesterification of tannic acid using propanol as organic reaction media under low water conditions. The identity of the product was confirmed with thin layer chromatography and Fourier transform infrared spectroscopy. It was noted that 699 U/ml of enzyme could give 60% solubilisation of tea cream within 1 h. Enzyme production medium was optimized adopting Box–Behnken design for simultaneous synthesis of tannase and gallic acid. Process variables including tannic acid, sodium chloride, ferrous sulphate, dipotassium hydrogen phosphate, incubation period and agitation were recognized as the critical factors that influenced tannase and gallic acid production. The model obtained predicted 4,824.61 U/ml of tannase and 136.206 μg/ml gallic acid after 48 h of incubation, whereas optimized medium supported 5,085 U/ml tannase and 372.6 μg/ml of gallic acid production after 36 and 84 h of incubation, respectively, with a 15-fold increase in both enzyme and gallic acid production. Results indicated scope for utilization of this acidophilic tannase for transesterification of tannic acid into propyl gallate, tea cream solubilisation and simultaneous production of gallic acid along with tannase

Valorization of date palm (Phoenix dactylifera) fruit processing by-products and wastes using bioprocess technology – Review

The date palm Phoenix dactylifera has played an important role in the day-to-day life of the people for the last 7000 years. Today worldwide production, utilization and industrialization of dates are continuously increasing since date fruits have earned great importance in human nutrition owing to their rich content of essential nutrients. Tons of date palm fruit wastes are discarded daily by the date processing industries leading to environmental problems. Wastes such as date pits represent an average of 10% of the date fruits. Thus, there is an urgent need to find suitable applications for this waste. In spite of several studies on date palm cultivation, their utilization and scope for utilizing date fruit in therapeutic applications, very few reviews are available and they are limited to the chemistry and pharmacology of the date fruits and phytochemical composition, nutritional significance and potential health benefits of date fruit consumption. In this context, in the present review the prospects of valorization of these date fruit processing by-products and wastes’ employing fermentation and enzyme processing technologies towards total utilization of this valuable commodity for the production of biofuels, biopolymers, biosurfactants, organic acids, antibiotics, industrial enzymes and other possible industrial chemicals are discussed

Fish flesh agar medium - a suitable experimental medium for the detection of spoilage bacteria

The spoilage characteristics of bacterial strains were studied by growing them at 28 _+ 2 °C in agar and broth media prepared with sterile fish and prawn flesh homogenates. The percentage of spoilers found among the bacterial isolates tested, as shown by odour production and halo zone formation, was independent of the source of flesh used. Indole and fluorescent pigment production were also observed in the broth. Pseudomonas, Vibrio and Acinetobacter exhibited faster growth in flesh media than in the usual artificial media. Decrease of protein and lipid concentration in the clear zone of agar media suggests the utilization of the available substrate by spoilage bacteria.

Trypsin Inhibitor from Edible Mushroom Pleurotus floridanus Active against Proteases of Microbial Origin

Protease inhibitors can be versatile tools mainly in the fields of medicine, agriculture and food preservative applications. Fungi have been recognized as sources of protease inhibitors, although there are only few such reports on mushrooms. This work reports the purification and characterization of a trypsin inhibitor from the fruiting body of edible mushroom Pleurotus floridanus (PfTI) and its effect on the activity of microbial proteases. The protease inhibitor was purified up to 35-fold by DEAE-Sepharose ion exchange column, trypsin-Sepharose column and Sephadex G100 column. The isoelectric point of the inhibitor was 4.4, and its molecular mass was calculated as 37 kDa by SDS-PAGE and 38.3 kDa by MALDI-TOF. Inhibitory activity confirmation was by dot-blot analysis and zymographic activity staining. The specificity of the inhibitor toward trypsin was with Ki of 1.043×10−10 M. The inhibitor was thermostable up to 90 °C with maximal stability at 30 °C, active over a pH range of 4–10 against proteases from Aspergillus oryzae, Bacillus licheniformis, Bacillus sp. and Bacillus amyloliquefaciens. Results indicate the possibility of utilization of protease inhibitor from P. floridanus against serine proteases

Selection of Marine Yeasts for the Generation of Single Cell Protein from Prawn-shell Waste

Marine yeasts (33 strains) were isolated from the coastal and offshore waters off Cochin. The isolates were identified and then characterized for the utilization of starch, gelatin, lipid, cellulose, urea, pectin, lignin, chitin and prawn-shell waste. Most of the isolates were Candida species. Based on the biochemical characterization, four potential strains were selected and their optimum pH and NaCI concentration for growth were determined. These strains were then inoculated into prawn-shell waste and SCP (single cell protein) generation was noted in terms of the increase in protein content of the final product.

Design and Development of a Remotely Operated Underwater Multi-Robot Manipulator Controller

For the scientific and commercial utilization of Ocean resources, the role of intelligent underwater robotic systems are of great importance. Scientific activities like Marine Bio-technology, Hydrographic mapping, and commercial applications like Marine mining, Ocean energy, fishing, aquaculture, cable laying and pipe lining are a few utilization of ocean resources. As most of the deep undersea exploration are beyond the reachability of divers and also as the use of operator controlled and teleoperated Remotely Operated Vehicles (ROVs) and Diver Transport Vehicles (DTVs) turn out to be highly inefficient, it is essential to have a fully automated system capable providing stable control and communication links for the unstructured undersea environment.

Assessment of Energy Conservation in Indian Cement Industry and Forecasting of Co2 Emissions

Cement industry ranks 2nd in energy consumption among the industries in India. It is one of the major emitter of CO2, due to combustion of fossil fuel and calcination process. As the huge amount of CO2 emissions cause severe environment problems, the efficient and effective utilization of energy is a major concern in Indian cement industry. The main objective of the research work is to assess the energy cosumption and energy conservation of the Indian cement industry and to predict future trends in cement production and reduction of CO2 emissions. In order to achieve this objective, a detailed energy and exergy analysis of a typical cement plant in Kerala was carried out. The data on fuel usage, electricity consumption, amount of clinker and cement production were also collected from a few selected cement industries in India for the period 2001 - 2010 and the CO2 emissions were estimated. A complete decomposition method was used for the analysis of change in CO2 emissions during the period 2001 - 2010 by categorising the cement industries according to the specific thermal energy consumption. A basic forecasting model for the cement production trend was developed by using the system dynamic approach and the model was validated with the data collected from the selected cement industries. The cement production and CO2 emissions from the industries were also predicted with the base year as 2010. The sensitivity analysis of the forecasting model was conducted and found satisfactory. The model was then modified for the total cement production in India to predict the cement production and CO2 emissions for the next 21 years under three different scenarios. The parmeters that influence CO2 emissions like population and GDP growth rate, demand of cement and its production, clinker consumption and energy utilization are incorporated in these scenarios. The existing growth rate of the population and cement production in the year 2010 were used in the baseline scenario. In the scenario-1 (S1) the growth rate of population was assumed to be gradually decreasing and finally reach zero by the year 2030, while in scenario-2 (S2) a faster decline in the growth rate was assumed such that zero growth rate is achieved in the year 2020. The mitigation strategiesfor the reduction of CO2 emissions from the cement production were identified and analyzed in the energy management scenarioThe energy and exergy analysis of the raw mill of the cement plant revealed that the exergy utilization was worse than energy utilization. The energy analysis of the kiln system showed that around 38% of heat energy is wasted through exhaust gases of the preheater and cooler of the kiln sysetm. This could be recovered by the waste heat recovery system. A secondary insulation shell was also recommended for the kiln in the plant in order to prevent heat loss and enhance the efficiency of the plant. The decomposition analysis of the change in CO2 emissions during 2001- 2010 showed that the activity effect was the main factor for CO2 emissions for the cement industries since it is directly dependent on economic growth of the country. The forecasting model showed that 15.22% and 29.44% of CO2 emissions reduction can be achieved by the year 2030 in scenario- (S1) and scenario-2 (S2) respectively. In analysing the energy management scenario, it was assumed that 25% of electrical energy supply to the cement plants is replaced by renewable energy. The analysis revealed that the recovery of waste heat and the use of renewable energy could lead to decline in CO2 emissions 7.1% for baseline scenario, 10.9 % in scenario-1 (S1) and 11.16% in scenario-2 (S2) in 2030. The combined scenario considering population stabilization by the year 2020, 25% of contribution from renewable energy sources of the cement industry and 38% thermal energy from the waste heat streams shows that CO2 emissions from Indian cement industry could be reduced by nearly 37% in the year 2030. This would reduce a substantial level of greenhouse gas load to the environment. The cement industry will remain one of the critical sectors for India to meet its CO2 emissions reduction target. India’s cement production will continue to grow in the near future due to its GDP growth. The control of population, improvement in plant efficiency and use of renewable energy are the important options for the mitigation of CO2 emissions from Indian cement industries

Biopulping of Paddy Straw by Pleurotus eous

Biopulping being less energy intensive, inexpensive and causing lesser pollution, can be a viable alternative to chemical and mechanical pulping in paper and pulp industry. In view of shrinking forest reserves, agricultural residues are considered as an alternative raw material for making paper and board. By suitable treatment agriwaste can be converted into substrate for mushroom cultivation. Mushrooms of Pleurotus sp. can preferentially remove lignin from agriwaste with limited degradation to cellulose. The present study examines utilization of Pleurotus eous for biopulping of paddy straw by solid substrate fermentation. SMS, the mushroom growing medium that results from cultivation process, is a good source of fibre and can be pulped easily. Ligninases present in SMS were able to reduce lignin content to nearly half the initial amount by 21st day of cultivation. Highest cellulose content (% dry weight) was observed on 21st day, while cellulase production commenced from 28th day of cultivation. SEM images revealed that SMS fibres are still associated with non-cellulosic materials when compared to chemically (20% w/v NaOH) extracted fibres.

Actinomycete Isolates from Arabian Sea and Bay of Bengal: Biochemical, Molecular and Functional Characterization

The thesis is comprised of seven chapters. Chapter 1 gives a general introduction to marine actinomycetes; Chapter 2 gives an account on the morphological, biochemical and physiological characterization of marine actinomycetes. Comprehensive description of molecular identification and phylogenetic analysis of actinomycetes is dealt with in Chapter 3. The antimicrobial property with special reference to antivibrio activity is described in Chapter 4. Chapter 5 explores the melanin production ability of marine actinomycetes, characterization of melanin and evaluation of its bioactivity. Chapter 6 illustrates the study on chitinolytic Streptomyces as antifungal and insecticidal agents. Summary and Conclusion of the study is presented in Chapter 7, followed by References and Appendices.The present study provides an insight into the various actinomycetes occurring in the sediments of Arabian Sea and Bay of Bengal. Streptomyces was found to be the dominant group followed by Nocardiopsis. Eventhough generic level identification is possible by traditional phenotypic methods, species level identification necessitate a polyphasic approach including both phenotypic and genotypic characterization. Antibiotic production coupled with biogranulation property helped in the effective utilization of the actinomycetes for the control of vibrios. Melanin from Streptomyces bikiniensis was proved to be a promising antioxidant and photoprotectant. Marine actinomycetes were found to be a good source of hydrolytic enzymes and the chitinolytic isolates could be explored as biocontrol agents in terms of antifungal and insecticidal property. The present study explored the potential of marine actinomycetes especially Streptomycetes as a promising source of bioactive molecules for application in aquaculture and pharmaceutical industry.

Characterization and Pathogenicity of Vibrio cholerae and Vibrio vulnificus from Marine environments

The genus Vibrioof the family Vibrionaceae are Gram negative, oxidasepositive, rod- or curved- rodshaped facultative anaerobes, widespread in marine and estuarine environments. Vibrio species are opportunistic human pathogens responsible for diarrhoeal disease, gastroenteritis, septicaemia and wound infections and are also pathogens of aquatic organisms, causing infections to crustaceans, bivalves and fishes. In the present study, marine environmental samples like seafood and water and sediment samples from aquafarms and mangroves were screened for the presence of Vibrio species. Of the134 isolates obtained from the various samples, 45 were segregated to the genus Vibrio on the basis of phenotypic characterization.like Gram staining, oxidase test, MoF test and salinity tolerance. Partial 16S rDNA sequence analysis was utilized for species level identification of the isolates and the strains were identified as V. cholerae(N=21), V. vulnificus(N=18), V. parahaemolyticus(N=3), V. alginolyticus (N=2) and V. azureus (N=1). The genetic relatedness and variations among the 45 Vibrio isolates were elucidated based on 16S rDNA sequences. Phenotypic characterization of the isolates was based on their response to 12 biochemical tests namely Voges-Proskauers’s (VP test), arginine dihydrolase , tolerance to 3% NaCl test, ONPG test that detects β-galactosidase activity, and tests for utilization of citrate, ornithine, mannitol, arabinose, sucrose, glucose, salicin and cellobiose. The isolates exhibited diverse biochemical patterns, some specific for the species and others indicative of their environmental source.Antibiogram for the isolates was determined subsequent to testing their susceptibility to 12 antibiotics by the disc diffusion method. Varying degrees of resistance to gentamycin (2.22%), ampicillin(62.22%), nalidixic acid (4.44%), vancomycin (86.66), cefixime (17.77%), rifampicin (20%), tetracycline (42.22%) and chloramphenicol (2.22%) was exhibited. All the isolates were susceptible to streptomycin, co-trimoxazole, trimethoprim and azithromycin. Isolates from all the three marine environments exhibited multiple antibiotic resistance, with high MAR index value. The molecular typing methods such as ERIC PCR and BOX PCR revealed intraspecies relatedness and genetic heterogeneity within the environmental isolatesof V. cholerae and V. vulnificus. The 21 strains of V. choleraewere serogroupedas non O1/ non O139 by screening for the presence O1rfb and O139 rfb marker genes by PCR. The virulence/virulence associated genes namely ctxA, ctxB, ace, VPI, hlyA, ompU, rtxA, toxR, zot, nagst, tcpA, nin and nanwere screened in V. cholerae and V. vulnificusstrains.The V. vulnificusstrains were also screened for three species specific genes viz., cps, vvhand viu. In V. cholerae strains, the virulence associated genes like VPI, hlyA, rtxA, ompU and toxR were confirmed by PCR. All the isolates, except for strain BTOS6, harbored at least one or a combination of the tested genes and V. choleraestrain BTPR5 isolated from prawn hosted the highest number of virulence associated genes. Among the V. vulnificusstrains, only 3 virulence genes, VPI, toxR and cps, were confirmed out of the 16 tested and only 7 of the isolates had these genes in one or more combinations. Strain BTPS6 from aquafarm and strain BTVE4 from mangrove samples yielded positive amplification for the three genes. The toxRgene from 9 strains of V. choleraeand 3 strains of V. vulnificus were cloned and sequenced for phylogenetic analysis based on nucleotide and the amino acid sequences. Multiple sequence alignment of the nucleotide sequences and amino acid sequences of the environmental strains of V. choleraerevealed that the toxRgene in the environmental strains are 100% homologous to themselves and to the V. choleraetoxR gene sequence available in the Genbank database. The 3 strains of V. vulnificus displayed high nucleotide and amino acid sequence similarity among themselves and to the sequences of V. cholerae and V. harveyi obtained from the GenBank database, but exhibited only 72% homology to the sequences of its close relative V. vulnificus. Structure prediction of the ToxR protein of Vibrio cholerae strain BTMA5 was by PHYRE2 software. The deduced amino acid sequence showed maximum resemblance with the structure of DNA-binding domain of response regulator2 from Escherichia coli k-12 Template based homology modelling in PHYRE2 successfully modelled the predicted protein and its secondary structure based on protein data bank (PDB) template c3zq7A. The pathogenicity studies were performed using the nematode Caenorhabditiselegansas a model system. The assessment of pathogenicity of environmental strain of V. choleraewas conducted with E. coli strain OP50 as the food source in control plates, environmental V. cholerae strain BTOS6, negative for all tested virulence genes, to check for the suitability of Vibrio sp. as a food source for the nematode;V. cholerae Co 366 ElTor, a clinical pathogenic strain and V. cholerae strain BTPR5 from seafood (Prawn) and positive for the tested virulence genes like VPI, hlyA, ompU,rtxA and toxR. It was found that V. cholerae strain BTOS6 could serve as a food source in place of E. coli strain OP50 but behavioral aberrations like sluggish movement and lawn avoidance and morphological abnormalities like pharyngeal and intestinal distensions and bagging were exhibited by the worms fed on V. cholerae Co 366 ElTor strain and environmental BTPR5 indicating their pathogenicity to the nematode. Assessment of pathogenicity of the environmental strains of V. vulnificus was performed with V. vulnificus strain BTPS6 which tested positive for 3 virulence genes, namely, cps, toxRand VPI, and V. vulnificus strain BTMM7 that did not possess any of the tested virulence genes. A reduction was observed in the life span of worms fed on environmental strain of V. vulnificusBTMM7 rather than on the ordinary laboratory food source, E. coli OP50. Behavioral abnormalities like sluggish movement, lawn avoidance and bagging were also observed in the worms fed with strain BTPS6, but the pharynx and the intestine were intact. The presence of multi drug resistant environmental Vibrio strainsthat constitute a major reservoir of diverse virulence genes are to be dealt with caution as they play a decisive role in pathogenicity and horizontal gene transfer in the marine environments.

Population characteristics and taxonomy of lantern fishes of genus Diaphus (Family Myctophidae) off south west coast of India

Globally most of the conventional fish stocks have reached a state of optimum exploitation or even over-exploitation; efficient utilization of non-conventional resources is necessary to meet the supply-demand gap for protein supply. Mesopelagic fishes can be considered as one such promising resource for the future, if appropriate harvest and post-harvest technologies are developed. Increasing human population and increasing demand for cheaper food fishes has made myctophids a possible potential resource for future exploitation and utilization. Earlier studies indicated the abundance of Diaphus spp. in the eastern and northeastern Arabian Sea. The present study also indicates the dominance of Diaphus spp. in the deep sea trawling grounds of south west coast of India. Commercial viability of the myctophid fishing in the Indian waters has to be worked out. The present catch estimation is based on the Stratified Random Sampling Method from the landing data. As the coverage of sampling area was limited and the gear efficiency was not standardized, the data generated are not precise. A counter check for the estimates is also not possible due to the absence of comparable works in the study area. Fish biomass estimation by acoustics survey coupled with direct fishing would only confirm the accuracy of estimates. Exploratory surveys for new fishing areas to be continued, for gathering the distribution, abundance, biological and ecological data and map the potential fishing ground on a GIS platform and the data should be provided to the commercial entrepreneurs. Generally non-conventional and non-targeted resources are under low fishing pressure and exploitation rates. Low values of fishing mortality and exploitation rates indicate that removal from the stock by fishing was only nominal from the present fishing grounds. The results indicate that the stock is almost at virgin state and remains grossly underexploited. Since the extent of distribution and abundance of the stock in the ecosystem remains to be ascertained, sustainable yield could not be estimated. Also the impact of myctophids harvest, on other commercially important fishes, has to be studied.

Investigations on the structural and operational changes of Ring Seine Fishing Systems of Kerala and its Life Cycle Assessment (lCA)

The study covers theFishing capture technology innovation includes the catching of aquatic animal, using any kind of gear techniques, operated from a vessel. Utilization of fishing techniques varies, depending upon the type of fisheries, and can go from a basic and little hook connected to a line to huge and complex mid water trawls or seines operated by large fishing vessels.The size and autonomy of a fishing vessel is largely determined by its ability to handle, process and store fish in good condition on board, and thus these two characteristics have been greatly influenced by the introduction and utilization of ice and refrigeration machinery. Other technological developments especially hydraulic hauling machinery, fish finding electronics and synthetic twines have also had a major impact on the efficiency and profitability of fishing vessels.A wide variety of fishing gears and practices ranging from small-scale artisanal to advanced mechanised systems are used for fish capture in Kerala. Most important among these fishing gears are trawls, seines, lines, gillnets and entangling nets and traps The modern sector was introduced in 1953 at Neendakara, Shakthikulangara region under the initiative of Indo-Norwegian project (INP). The novel facilities introduced in fishing industry by Indo- Norwegian project accordingly are mechanically operated new boats with new fishing nets. Soon after mechanization, motorization programme gained momentum in Kerala especially in Alleppey, Ernakulam and Kollam districts.