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 .

Biocompatible polyhydroxybutyrate (PHB) production by marine Vibrio azureus BTKB33 under submerged fermentation

Polyhydroxybutyrate (PHB) is known to have applications as medical implants and drug delivery carriers and is consequently in high demand. In the present study the possibilities of harnessing potential PHB-producing vibrios from marine sediments as a new source of PHB was investigated since marine environments are underexplored. Screening of polyhydroxyalkanoate (PHA)-producing vibrios from marine sediments was performed using a fluorescent plate assay followed by spectrophotometric analysis of liquid cultures. Out of 828 isolates, Vibrio sp. BTKB33 showed maximum PHA production of 0.21 g/L and PHA content of 193.33 mg/g of CDW. The strain was identified as Vibrio azureus based on phenotypic characterization and partial 16S rDNA sequence analysis. The strain also produced several industrial enzymes: amylase, caseinase, lipase, gelatinase, and DNase. The FTIR analysis of extracted PHA and its comparison with standard PHB indicated that the accumulated PHA is PHB. Bioprocess development studies for enhancing PHA production were carried out under submerged fermentation conditions. Optimal submerged fermentation conditions for enhanced intracellular accumulation of PHA production were found to be 35 °C, pH −7, 1.5 % NaCl concentration, agitation at 120 rpm, 12 h of inoculum age, 2.5 % initial inoculum concentration, and 36 h incubation along with supplementation of magnesium sulphate, glucose, and ammonium chloride. The PHA production after optimization was found to be increased to 0.48 g/L and PHA content to426.88 mg/g of CDW, indicating a 2.28-fold increase in production. Results indicated that V. azureus BTKB33 has potential for industrial production of PHB.

Polyphosphate Accumulation by Marine Bacteria

Phosphate (Pi) is one among the most important essential residues in maintenance and inheritance of life, with far diverse physiological role as structural, functional and energy transduction. Phosphate accumulation in wastewaters containing run off of fertilizers and industrial discharges is a global problem that results in algal blooms in bays, lakes and waterways. Currently available methods for removing phosphates from wastewater are based primarily on polyP accumulation by the activated sludge bacteria. PolyP plays a critical role in several environmental and biotechnological problems. Possible relation of interaction between polyP accumulation phenomenon, the low biomass, low Pi uptake, and varying results obtained in response to the impact of sodium chloride, pH, temperature, various inorganic salts and additional carbon sources studied, are all intriguing observations in the present investigation. The results of the present study have evidenced very clearly the scope for potential strains of bacteria from both sea water and marine sediments which could be exploited both for Pi removal in wastewater released by industries and intensive aquaculture practices in to the aquatic environment as well as to harness the potential strains for industrial production of polyP which was wide range of applications.

L-Glutaminase production by an immobilized marine Pseudomonas sp. BTMS -51

The present study is about the Pseudomonas sp. BTMS-51 isolated from the marine sediments of Cochin Coast. In the present study, it is concluded that marine bacteria are ideal candidates for immobilization using either Ca-alginate entrapment or physical adsorption on to synthetic inert supports and the process of immobilization does not negatively influence them. Thus, Ca-alginate entrapment of the bacteria was found to be well suited for reuse of the biomass and extended operational stability during continuous operation. Adherence of the bacterium to inertsupports was observed to be strong and it imparted minimal stress on the immobilized bacterium and allowed detachment and relocation on the supports which enabled the formation of a dynamic equilibrium maintaining a stable cell loading. This is particularly desirable in the industry for extended operational stability and maintenance of consistently higher outputs. Marine Pseudomonas sp. BTMS-51 is ideal for industrial production of extra cellular L-glutaminase and immobilization on to synthetic inert support such as polyurethane foam could be an efficient technique, employing packed bed reactor for continuous production of the enzyme. Temperature and glutamine concentration had significant effects on enzyme production by cells immobilized on polyurethane foam (PUF).

Heterotrophic bacteria from the continental slope sediments of arabian sea

The marine environment is indubitably the largest contiguous habitat on Earth. Because of its vast volume and area, the influence of the world ocean on global climate is profound and plays an important role in human welfare and destiny. The marine environment encompasses several habitats, from the sea surface layer down through the bulk water column, which extends >10,000 meters depth, and further down to the habitats on and under the sea floor. Compared to surface habitats, which have relatively high kinetic energy, deep-ocean circulation is very sluggish. By comparison, life in the deep sea is characterized by a relatively constant physical and chemical environment. Deep water occupying the world ocean basin is a potential natural resource based on its properties such as low temperature, high pressure and relatively unexplored properties. So, a judicious assessment of the marine resources and its management are essential to ensure sustainable development of the country’s ocean resources. Marine sediments are complex environments that are affected by both physiological and biological factors, water movements and burrowing animals. They encompass a large extent of aggregates falling from the surface waters. In aquatic ecosystems, the flux of organic matter to the bottom sediments depend on primary productivity at the ocean surface and water depth. Over 50% of the earth’s surface is covered by deep-sea sediments that are primarily formed through the continual deposition of particles from the productive pelagic waters (Vetriani et al., 1999). These aggregates are regarded as ‘hot spots’ of microbial activity in the ocean (Simon et al., 2002). This represents a good nutritional substrate for heterotrophic bacteria and favours bacterial growth