Anirban Roy Choudhury

Exploitation of marine bacteria for production of gold nanoparticles

BackgroundGold nanoparticles (AuNPs) have found wide range of applications in electronics, biomedical engineering, and chemistry owing to their exceptional opto-electrical properties. Biological synthesis of gold nanoparticles by using plant extracts and microbes have received profound interest in recent times owing to their potential to produce nanoparticles with varied shape, size and morphology. Marine microorganisms are unique to tolerate high salt concentration and can evade toxicity of different metal ions. However, these marine microbes are not sufficiently explored for their capability of metal nanoparticle synthesis. Although, marine water is one of the richest sources of gold in the nature, however, there is no significant publication regarding utilization of marine micro-organisms to produce gold nanoparticles. Therefore, there might be a possibility of exploring marine bacteria as nanofactories for AuNP biosynthesis.ResultsIn the present study, marine bacteria are exploited towards their capability of gold nanoparticles (AuNPs) production. Stable, monodisperse AuNP formation with around 10 nm dimension occur upon exposure of HAuCl4 solution to whole cells of a novel strain of Marinobacter pelagius, as characterized by polyphasic taxonomy. Nanoparticles synthesized are characterized by Transmission electron microscopy, Dynamic light scattering and UV-visible spectroscopy.ConclusionThe potential of marine organisms in biosynthesis of AuNPs are still relatively unexplored. Although, there are few reports of gold nanoparticles production using marine sponges and sea weeds however, there is no report on the production of gold nanoparticles using marine bacteria. The present work highlighted the possibility of using the marine bacterial strain of Marinobacter pelagius to achieve a fast rate of nanoparticles synthesis which may be of high interest for future process development of AuNPs. This is the first report of AuNP synthesis by marine bacteria.

Biosynthesis of gold and silver nanoparticles using a novel marine strain of Stenotrophomonas

The present study aims at exploiting marine microbial diversity for biosynthesis of metal nanoparticles and also investigates role of microbial proteins in the process of bio-mineralization of gold and silver. This is the first report for concurrent production of gold and silver nanoparticles (AuNPs and AgNPs) by extracellular secretion of a novel strain of Stenotrophomonas, isolated from Indian marine origin. This novel strain has faster rate kinetics for AgNPs synthesis than any other organism reported earlier. The nanoparticles were further characterized using UV-vis spectrophotometer, TEM, DLS and EDAX confirming their size ranging from 10-50 nm and 40-60 nm in dimensions for AuNPs and AgNPs, respectively. TEM analysis indicated formation of multi-shaped nanoparticles with heterogeneous size distribution in both the cases. Finally, the SDS-PAGE analysis of extracellular media supernatant suggested a potential involvement of certain low molecular weight secretory proteins in AuNPs and AgNPs biosynthesis.

Utilization of corn steep liquor for biosynthesis of pullulan, an important exopolysaccharide.

Five different agricultural wastes viz. rice bran oil cake, soya bean oil cake, cotton seed oil cake, mustard seed oil cake and corn steep liquor (CSL) were evaluated for their use as nutrient along with 15% (w/v) glucose as carbon source for biosynthesis of pullulan using Aureobasidium pullulans RBF 4A3. Among the selected agricultural wastes, CSL was found to be the best and supported production of 77.92gL(-1) pullulan under un-optimized conditions. Single point optimization technique resulted in increase in 18% pullulan (88.59gL(-1)) production. The process was successfully validated in a 7-L fermenter and a process economic analysis has suggested that use of CSL as nutrient may result in 3-fold reduction of cost of raw materials for pullulan production as compared to a process where conventional nitrogen sources were used. These observations may be helpful in development of a cost effective process for pullulan production.

Extracellular Polysaccharide Production by a Novel Osmotolerant Marine Strain of Alteromonas macleodii and Its Application towards Biomineralization of Silver

The present study demonstrates exopolysaccharide production by an osmotolerant marine isolate and also describes further application of the purified polysaccharide for production of colloidal suspension of silver nanoparticles with narrow size distribution. Phylogenetic analysis based on 16S r RNA gene sequencing revealed close affinity of the isolate to Alteromonas macleodii. Unlike earlier reports, where glucose was used as the carbon source, lactose was found to be the most suitable substrate for polysaccharide production. The strain was capable of producing 23.4 gl−1 exopolysaccharide with a productivity of 7.8 gl−1 day−1 when 15% (w/v) lactose was used as carbon source. Furthermore, the purified polysaccharide was able to produce spherical shaped silver nanoparticles of around 70 nm size as characterized by Uv-vis spectroscopy, Dynamic light scattering and Transmission electron microscopy. These observations suggested possible commercial potential of the isolated strain for production of a polysaccharide which has the capability of synthesizing biocompatible metal nanoparticle.

Cost effective production of pullulan from agri-industrial residues using response surface methodology.

Response surface methodology was used to develop an economically feasible process for the fermentative production of pullulan using agri-industrial residues, jaggery, de-oiled jatropha seed cake (DOJSC) and corn steep liquor (CSL), as sole media components. A second order polynomial model was obtained using central composite design to understand the effects of interactions among these substrates on pullulan biosynthesis. Results indicated that, lower concentrations of CSL and DOJSC and higher concentrations of jaggery favoured pullulan production. The optimal nutrient composition (18% jaggery, 3% DOJSC and 0.97% CSL) as suggested by the model resulted in production of 66.25 g/L pullulan with a productivity of 0.92 g/Lh. Analysis of raw material cost component for pullulan production suggested that sole utilization of agri-residues may lead to development of cost effective process for pullulan production.

Deoiledjatropha seed cake is a useful nutrient for pullulan production

BackgroundEver increasing demand for fossil fuels is a major factor for rapid depletion of these non-renewable energy resources, which has enhanced the interest of finding out alternative sources of energy. In recent years jatropha seed oil has been used extensively for production of bio-diesel and has shown significant potential to replace petroleum fuels at least partially. De-oiled jatropha seed cake (DOJSC) which comprises of approximately 55 to 65% of the biomass is a byproduct of bio-diesel industry. DOJSC contains toxic components like phorbol esters which restricts its utilization as animal feed. Thus along with the enhancement of biodiesel production from jatropha, there is an associated problem of handling this toxic byproduct. Utilization of DOJSC as a feed stock for production of biochemicals may be an attractive solution to the problem.Pullulan is an industrially important polysaccharide with several potential applications in food, pharmaceuticals and cosmetic industries. However, the major bottleneck for commercial utilization of pullulan is its high cost. A cost effective process for pullulan production may be developed using DOJSC as sole nutrient source which will in turn also help in utilization of the byproduct of bio-diesel industry.ResultsIn the present study, DOJSC has been used as a nutrient for production of pullulan, in place of conventional nutrients like yeast extract and peptone. Process optimization was done in shake flasks, and under optimized conditions (8% DOJSC, 15% dextrose, 28°C temperature, 200 rpm, 5% inoculum, 6.0 pH) 83.98 g/L pullulan was obtained. The process was further validated in a 5 L laboratory scale fermenter.ConclusionThis is the first report of using DOJSC as nutrient for production of an exopolysaccharide. Successful use of DOJSC as nutrient will help in finding significant application of this toxic byproduct of biodiesel industry. This in turn also have a significant impact on cost reduction and may lead to development of a cost effective green technology for pullulan production.

Multi-analytical approach to understand biomineralization of gold using rice bran: a novel and economical route

Biomimetic fabrication of gold nanoparticles (AuNP) has received significant attention worldwide. However, most of the reports were unable to clearly indicate the responsible biomolecule involved in the production of AuNP. The present study aimed to address this challenge by critically identifying the active component of rice bran responsible for the biomineralization of gold. Fractionation of rice bran extract by HPLC and further characterization using LC-MS revealed that ferulic acid is the functional molecule responsible for the bioconversion of Au3+ to Au0. Real time monitoring of heat changes by isothermal titration calorimetry indicated biosynthesis of AuNP was exothermic in nature. Additionally, reaction between ferulic acid and HAuCl4 for AuNP formation was observed to follow first order kinetics and finally various thermodynamic parameters were estimated by measuring residual ferulic acid concentration. Combinatorial observation of all these studies might lead the way to decipher the exact mechanism of nanoparticle biosynthesis.

Understanding the effect of interaction among aeration, agitation and impeller positions on mass transfer during pullulan fermentation by Aureobasidium pullulans

Pullulan is a non-ionic, water-soluble homopolysaccharide produced via fermentation using Aureobasidium pullulans, a black yeast. The unique physicochemical properties of pullulan have made it a potential candidate for a diverse range of applications in various industrial sectors such as food, pharmaceutical, cosmetics, and biomedicine. Low yield and productivity are major challenges for the extensive commercialization of this biopolymer, and they are often found to be associated with poor mass transfer during the fermentative production of pullulan. The present study is an attempt to address this challenge by a unique multivariate approach. The interactive influence of air-flow rates, agitation speed and impeller spacing on volumetric mass transfer coefficient (kLa) was investigated using a statistical design. The design has demonstrated a unique correlation between mass transfer capabilities and process variables, whereas pullulan fermentation kinetics has revealed a dependence of microbial activity on kLa values in a bioprocess. Furthermore, the resultant mass transfer conditions were applied for pullulan production, which enhanced the productivity significantly (1.61 g L−1 h−1). These observations might help in obtaining improved mass transfer conditions for the economic production of pullulan.

Facile and rapid thermo-regulated biomineralization of gold by pullulan and study of its thermodynamic parameters

A novel method for the production of gold nanoparticles (AuNPs) using pullulan as reducing and stabilizing agent has been developed. Quasi-spherical shaped AuNPs in the range of 50-100 nm were produced at three different temperature regimes 80°C, 90°C and 100°C as characterized using UV-vis spectrophotometer, TEM and DLS. Study of reaction kinetics and thermodynamic parameters indicated that the reaction between pullulan and chloroauric acid for AuNPs formation followed first order reaction kinetics and higher temperature was favorable for the synthesis of smaller sized AuNPs. FT-IR data analyses, provided an insight towards the mechanism of gold nanoparticle formation which suggested that, the free CH2OH groups of pullulan molecule were oxidized to carboxylate ions resulted in formation of AuNPs whereas the basic skeletal structure of pullulan remained unaltered. This study may open up new avenues for synthesis of tailor made biogenic AuNPs with possible application in biomedical field.

Development of suitable solvent system for downstream processing of biopolymer pullulan using response surface methodology.

Downstream processing is an important aspect of all biotechnological processes and has significant implications on quality and yield of the final product. Several solvents were examined for their efficacy on pullulan precipitation from fermentation broth. Interactions among four selected solvents and their effect on pullulan yield were studied using response surface methodology. A polynomial model was developed using D-optimal design and three contour plots were generated by performing 20 different experiments and the model was validated by performing optimization experiments. The results indicated that lower concentration of ethanol in combination with the other three solvents has resulted in higher yield of polymer from fermentation broth and the optimized solvent system was able to recover 1.44 times more pullulan as compared to the conventional ethanolic precipitation method. These observations may help in enhancing efficiency of pullulan recovery from fermentation broth and also result in reduced cost of production for the final product.