Characterization of polyhydroxyalkanoates accumulating vibrios from marine benthic environments and production studies of polyhydroxyalkanoates by Vibrio sp. BTKB33

The present work deals with the characterization of polyhydroxyalkanoates accumulating vibrios from marine benthic environments and production studies of polyhydroxyalkanoates by vibrio sp.BTKB33. Vibrios are a group of (iram negative, curved or straight motile rods that normally inhabit the aquatic environments.The present study therefore aimed at evaluating the occurrence of PHA accumulating vibrios inhabiting marine benthic environments; characterizing the potential PHA accumulators employing phenotypic and genotypic approaches and molecular characterization of the PHA synthase gene. The study also evaluated the PHA production in V:'hri0 sp. strain BTKB33, through submerged fennentation using statistical optimization and characterized the purified biopolymer. Screening for PHA producing vibrios from marine benthic environments. Characterization of PHA producers employing phenotypic and genotypic approaches.The incidence of PHA accumulation in Vibrio sp. isolated from marine sediments was observed to be high, indicating that the natural habitat of these bacteria are stressful. Considering their ubiquitous nature, the ecological role played by vibrios in maintaining the delicate balance of the benthic ecosystem besides returning potential strains, with the ability to elaborate a plethora of extracellular enzymes for industrial application, is significant. The elaboration of several hydrolytic enzymes by individuals also emphasize the crucial role of vibrios in the mineralization process in the marine environment. This study throws light on the extracellular hydrolytic enzyme profile exhibited by vibrios. It was concluded that apart from the PHA accumulation, presence of exoenzyme production and higher MAR index also aids in their survival in the highly challenging benthic enviromnents. The phylogenetic analysis of the strains and studies on intra species variation within PHA accumulating strains reveal their diversity. The isolate selected for production in this study was Vibrio sp. strain BTKB33, identified as V.azureus by 16S rDNA sequencing and phenotypic characterization. The bioprocess variables for PHA production utilising submerged fermentation was optimized employing one-factor-at-a-time-method, PB design and RSM studies. The statistical optimization of bioprocess variables revealed that NaCl concentration, temperature and incubation period are the major bioprocess variables influencing PHA production and PHA content. The presence of Class I PHA synthase genes in BTKB33 was also unveiled. The characterization of phaC genes by PCR and of the extracted polymer employing FTIR and NMR analysis revealed the presence of polyhydroxybutyrate, smallest known PI-IAs, having wider domestic, industrial and medical application. The strain BTKB33 bearing a significant exoenzyme profile, can thus be manipulatedin future for utilization of diverse substrates as C- source for PHA production. In addition to BTKB33, several fast growing Vibrio sp. having PHA accumulating ability were also isolated, revealing the prospects of this environment as a mine for novel PHA accumulating microbes. The findings of this study will provide a reference for further research in industrial production of PHAs from marine microorganisms .

Investigations on Phaeocystis sp., A Harmful Bloom- Forming Alga Isolated from Cochin Estuary

Considering the importance of diversity of micro algae in our ecosystem and new invasion of many organisms, an attempt was made to monitor the Cochin estuary along the south west coast of India for the qualitative distribution of phytoplankton and to study the growth kinetics and allelopathic effect of the phaeocystis sp. Isolated from the cochin estuary. Phaeocystis blooms are common only in high latitude environments and they rarely occur in low latitude environments such as tropics and subtropics. As phaeocystis is grouped under harmful alga ,in the present study the factors causing the blooms formation in the ecosystem. The nutrient concentration of the water body along with other physiochemical parameters that includes temperature salinity and ph play an important role in triggering the bloom of phaeocystis .The phaeocystis harbor specific bacterial flora associated with it and they exert an important role in the growth ,haemolytic activity and the bloom phases of the alga. The harmful alga mainly depends on the production of alleopathic compounds for the establishment of bloom in the marine environments .These physiological properties of the phaecystis were considered for the study, along with the role of nutrients in the allelopathic and hemolytic activity

Studies on L-glutamic acid Production using Brevibacterium sp.

The thesis comprises a set of experiments mainly focused on the improvement of L-glutamic acid fennentation. Much attention has been given to use of locally available raw materials, culturing the organism on inert solid substrates and also immobilization of the bacterial cells from the view point of long term utilization of biocatalyst and continuous operation of the stabilized system. Studies were also carried out for the down stream processing for the extraction and purification of L-glutamic acid. An attempt was made to study the morphological features of the microorganism including the cell premeability. In relation with the accumulation of glutamic acid within the cells an approach was made to study the behaviour of the Brevibacterium cells when they are exposed to hyper osmotic environment. Attempts were also made to study the requirement of iron and production of siderophores by this microbial strain. The search for a suitable nitrogen source for glutamate fermentation ended with a promising result that they got a potent urease activity and it can be utilized for many biotransfonnation studies. The entire thesis is presented in three sections, viz. introductory section, experimental section and the concluding section

Fermentation Of Rice And Related Products By Bacillus Sp.

Recently’ recognition cnf immobilization ‘technology for the rapid conversion of several substrates into metabolites and repeated reuse of the biocatalysts have drawn the attention of the fermentation scientists and technologists to try these new technologies for the rapid production of pnxkmt and enhancement of the efficiencies of the systems Hence in the present study rice was selected,as a substrate since it is a rich source of starch, available and cultivated throughout the year almost in all part of our country. Rice although known for its use as a staple food in many forms as rice, idli, dosai etc., has not been used in industry extensively. However, it ii; a potential resource for’ the production of alcohol, high protein food anui for sugar and sugar syrups as it is evidenced by the few reports mentioned in the review of literature. Of the several microorganisms available, Bacillus sp, is a known candidate for the production of amylases. Hence in the present study Bacillus sp, was desired for its known efficiencies in starch conversion

“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

Studies on Halocin production by haloarchaea Natrinema sp. BTSH10

This study on halocin production by Natrinema sp.BTSH10 indicate the prospects for intensive research which could lead to discovery of novel halocins which could have far reaching impact in biopharmaceutical industry particularly as anticancer drug. It is also anticipated that further research on this halocin could lead towards development of novel anticancer drug and new era in pharmaceutical biotechnology. There is no doubt that haloarchaea from saltern ponds have immense potential to return novel and valuable drugs and bioactive substances.

Bioremediation of Zinc Using Bacillus sp. Isolated from Metal- Contaminated Industrial Zone

The production of heavy metals has increased quickly since the industrial revolution. Heavy metals frequently form compounds that can be toxic, carcinogenic, or mutagenic, even in very small concentrations. The usual techniques of removing metals from wastewaters are in general expensive and have many restrictions. Alternative methods of metal removal and recovery based on biological materials have been measured. Among various agents, the use of microbes for the removal of metals from industrial and municipal wastewater has been proposed as a promising alternative to conventional heavy metal management strategies in past decades. Thus, the present study aims to isolate and characterize bacteria from soil, sediment, and waters of metal-contaminated industrial area to study the zinc resistance patterns and the zinc bioaccumulation potential of the selected microorganism. Zinc analysis of the samples revealed that concentrations varying from 39.832 m g/L to 310.24 m g/L in water, 12.81 m g/g to 407.53 m g/g in soil, and 81.06 m g/g to 829.54 m g/g in sediment are present. Bacterial zinc resistance study showed that tolerance to Zn was relatively low (<500 m g/ml). Ten bacterial genera were represented in soil and 11 from water, while only 5 bacterial genera were recorded from sediment samples. Bacillus, Pseudomonas , and Enterobacter were found in soil, sediment, and water samples. Highly zincresistant Bacillus sp. was selected for zinc removal experiment. Zinc removal studies revealed that at pH 5 about 40% reduction occurs; at pH 7, 25% occurs; and at pH 9, 50% occurs. Relatively an increased removal of Zinc was observed in the fi rst day of the experiment by Bacillus sp. The metal bioaccumulative potential of the selected isolates may have possible applications in the removal and recovery of zinc from industrial ef fluents.

Alkaline protease from a non-toxigenic mangrove isolate of Vibrio sp. V26 with potential application in animal cell culture

Vibrio sp. V26 isolated from mangrove sediment showed 98 % similarity to 16S rRNA gene of Vibrio cholerae, V. mimicus, V. albensis and uncultured clones of Vibrio. Phenotypically also it resembled both V. cholerae and V. mimicus.Serogrouping, virulence associated gene profiling, hydrophobicity, and adherence pattern clearly pointed towards the non—toxigenic nature of Vibrio sp. V26. Purification and characterization of the enzyme revealed that it was moderately thermoactive, nonhemagglutinating alkaline metalloprotease with a molecular mass of 32 kDa. The application of alkaline protease from Vibrio sp. V26 (APV26) in sub culturing cell lines (HEp-2, HeLa and RTG-2) and dissociation of animal tissue (chick embryo) for primary cell culture were investigated. The time required for dissociation of cells as well as the viable cell yield obtained by while administeringAPV26 and trypsin were compared. Investigations revealed that the alkaline protease of Vibrio sp. V26 has the potential to be used in animal cell culture for subculturing cell lines and dissociation of animal tissue for the development of primary cell cultures, which has not been reported earlier among metalloproteases of Vibrios.

L-Glutaminase production by marine Beau6eria sp. under solid state fermentation

Extracellular L-glutaminase production by Beau6eria sp., isolated from marine sediment, was observed during solid state fermentation using polystyrene as an inert support. Maximal enzyme production (49.89 U:ml) occurred at pH 9.0, 27°C, in a seawater based medium supplemented with L-glutamine (0.25% w:v) as substrate and D-glucose (0.5% w:v) as additional carbon source, after 96 h of incubation. Enzyme production was growth associated. Results indicate scope for production of salt tolerant L-glutaminase using this marine fungus

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

Biological coagulation of skim latex using Acinetobacter sp. isolated from natural rubber latex centrifugation effluent

An Acinetobacter sp, isolated from latex centrifugation effluent, effectively coagulated skim rubber from skim latex. After coagulation for 48 h without the addition of any nutrients, at an optimum dilution of 1:10(v/v) and with an inoculum concentration of 6.4 mg dry cell /ml, the yield of the skim rubber was 8 % (w/v) and the COD of the residual solution was only 0.4 g/l. chemical coagulation at the same dilution resulted in 7 % (w/v) yield of dry rubber content and 2.2 g COD /l.

A Novel Acinetobacter Sp. For 'Treating Highly Acidic Rubber Latex Cen'trifugation Effluent

A novel Acinetobacter sp. BTJR-IO isolated from highly acidic (pH 2.5-4.5) rubber latex centrifugation effluent with high COD (22000 rng/L) and BOD (5000 rng/L). This strain could effect 39.5% COD reduction on free cell inoculation of effluent without incorporation of additional nutrients after 8 days. CalciLnn alginate irrmobilized cells showed 16.4% and 25% COD reduction after 6 hra, without aeration and after 1 hr. with mild aeration under batch process respectively. Whereas 44.0% COD reduction could be achieved after 6 hrs. on continuous treatment in a packed bed reactor with mild aeration. Further, even after 3 cycles 37% COD reduction was recorded with continuous treatment

Halocin SH10 production by an extreme haloarchaeon Natrinema sp. BTSH10 isolated from salt pans of South India

Halobacteria, members of the domain Archaea that live under extremely halophilic conditions, are often considered as dependable source for deriving novel enzymes, novel genes, bioactive compounds and other industrially important molecules. Protein antibiotics have potential for application as preserving agents in food industry, leather industry and in control of infectious bacteria. Halocins are proteinaceous antibiotics synthesized and released into the environment by extreme halophiles, a universal characteristic of halophilic bacteria. Herein, we report the production of halocin (SH10) by an extremely halophilic archeon Natrinema sp. BTSH10 isolated from salt pan of Kanyakumari, Tamilnadu, India and optimization of medium for enhanced production of halocin. It was found that the optimal conditions for maximal halocin production were 42 C, pH 8.0, and 104 h of incubation at 200 rpm with 2% (V/V) inoculum concentration in Zobell’s medium containing 3 M NaCl, Galactose, beef extract, and calcium chloride as additional supplements. Results indicated scope for fermentation production of halocin for probable applications using halophilic archeon Natrinema sp. BTSH10

Characterization of Linear Low-Density Polyethylene/ Poly(vinyl alcohol) Blends and Their Biodegradability by Vibrio sp. Isolated from Marine Benthic Environment

Increasing amounts of plastic waste in the environment have become a problem of gigantic proportions. The case of linear low-density polyethylene (LLDPE) is especially significant as it is widely used for packaging and other applications. This synthetic polymer is normally not biodegradable until it is degraded into low molecular mass fragments that can be assimilated by microorganisms. Blends of nonbiodegradable polymers and biodegradable commercial polymers such as poly (vinyl alcohol) (PVA) can facilitate a reduction in the volume of plastic waste when they undergo partial degradation. Further, the remaining fragments stand a greater chance of undergoing biodegradation in a much shorter span of time. In this investigation, LLDPE was blended with different proportions of PVA (5–30%) in a torque rheometer. Mechanical, thermal, and biodegradation studies were carried out on the blends. The biodegradability of LLDPE/PVA blends has been studied in two environments: (1) in a culture medium containing Vibrio sp. and (2) soil environment, both over a period of 15 weeks. Blends exposed to culture medium degraded more than that exposed to soil environment. Changes in various properties of LLDPE/PVA blends before and after degradation were monitored using Fourier transform infrared spectroscopy, a differential scanning calorimeter (DSC) for crystallinity, and scanning electron microscope (SEM) for surface morphology among other things. Percentage crystallinity decreased as the PVA content increased and biodegradation resulted in an increase of crystallinity in LLDPE/PVA blends. The results prove that partial biodegradation of the blends has occurred holding promise for an eventual biodegradable product

Pretreatment Of Agricultural Waste With Pleurotus Sp. For Ethanol Production

Bioethanol is a liquid fuel obtained from fermentation of sugar/starch crops. Lignocellulosic biomass being less expensive is considered a future alternative for the food crops. One of the main challenges for the use of lignocellulosics is the development of an efficient pre-treatment process. Pretreatments are classified into three - physical, chemical, and biological pretreatment. Chemical process has not been proven suitable so far, due to high costs and production of undesired by-products. Biologically, hydrolysis can be enhanced by microbial or enzymatic pretreatment. Studies show that the edible mushrooms of Pleurotus sp. produce several extracellular enzymes which reduce the structural and chemical complexity of fibre. In the present study, P. ostreatus and P. eous were cultivated on paddy straw. Spent substrate left after mushroom cultivation was powdered and used for ethanol production. Saccharomyces sp. was used for fermentation studies. Untreated paddy straw was used as control. Production of ethanol from P. ostreatus substrate was 5.5 times more when compared to untreated paddy straw, while the spent substrate of P. eous gave 5 times increase in ethanol yield. Assays showed the presence of several extracellular enzymes in the spent substrate of both species, which together contributed to the increase in ethanol yield