Datta Madamwar

Sardar Patel University, BRD School of Biosciences, Faculty Member

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Papers by Datta Madamwar

Optimization of medium components for increased production of C-phycocyanin from Phormidium ceylanicum and its purification by single step process

Bioresource Technology, Feb 1, 2009

Phycocyanin is a major protein produced by cyanobacteria, but very few phycocyanin-producing stra... more Phycocyanin is a major protein produced by cyanobacteria, but very few phycocyanin-producing strains have been reported. In the present study, response surface methodology (RSM) involving a central composite design for four factors was successfully employed to optimize medium components for increased production of phycocyanin from Phormidium ceylanicum. The production of phycocyanin and interactions between sodium nitrate, calcium chloride, trace metal mix and citric acid stock were investigated and modeled. Under optimized condition P. ceylanicum was able to give 2.3-fold increase in phycocyanin production in comparison to commonly used BG 11 medium in 32 days. We have demonstrated the extraction, purification and characterization of C-phycocyanin using novel method based on filtration and single step chromatography. The protein was extracted by repeated freeze-thaw cycles and the crude extract was filtered and concentrated in stirred ultrafiltration cell (UFC). The UFC concentrate was then subjected to a single ion exchange chromatographic step. A purity ratio of 4.15 was achieved from a starting value of 1.05. The recovery efficiency of C-phycocyanin from crude extract was 63.50%. The purity was checked by electrophoresis and UV-Vis spectroscopy.

Stability of arginase in reverse micelles

Biotechnology Techniques, 1995

ABSTRACT

Activation and stabilization of invertase entrapped into reversed micelles of sodium lauryl sulfate and sodium tauroglycocholate in organic solvents

Enzyme and microbial technology, May 1, 1988

Abstract We report our observations on the activity of invertase enzyme entrapped inside the reve... more Abstract We report our observations on the activity of invertase enzyme entrapped inside the reversed micelles formed by either sodium tauroglycocholate or sodium lauryl sulfate in various organic solvents, namely hexane, n-decanol, benzene and carbon tetra-chloride. In this case more than four-fold increase in activity was observed under mild conditions in comparison to the activity in usual aqueous medium. Reversed micelles entrapped invertase exhibited higher velocity, low K M value and longer active life as compared to enzyme in aqueous phase. This is in support of the view that surfactant stabilizes active conformation of enzyme by entrapping them. The interior of the reversed micelle acts like a microreactor that provides favorable aqueous microenvironment for the enzyme activity.

Purification, characterization and comparison of phycoerythrins from three different marine cyanobacterial cultures

Bioresource Technology, 2011

The present study is focused on purification, characterization and comparison of phycoerythrins f... more The present study is focused on purification, characterization and comparison of phycoerythrins from three different marine cyanobacterial cultures--hormidium sp. A27 DM, Lyngbya sp. A09 DM and Halomicronema sp. A32 DM. &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39;Phycoerythrin&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39; was successfully purified and characterized. On SDS-PAGE, the PE purified from all three young cultures showed four bands--corresponding to α and β subunits of each of PE-I and PE-II. However, phycoerythrin purified after prolonged growth of Phormidium sp. A27 DM and Halomicronema sp. A32DM showed only one band corresponding to 14 kDa whereas Lyngbya sp. A09 DM continued to produce uncleaved phycoerythrin. The absorption spectra of purified PEs from all the three young and old cultures showed variations however the fluorescence studies of the purified PEs in all cases gave the emission spectra at around 580 nm. The described work is of great importance to understand the role of phycoerythrin in adapting cyanobacteria to stress conditions.

Naphthalene degradation by Pseudomonas sp. HOB1: In vitro studies and assessment of naphthalene degradation efficiency in simulated microcosms

Journal of Hazardous Materials, Jul 30, 2009

Naphthalene, being a ubiquitous pollutant of the environment and a perilous material, its biodegr... more Naphthalene, being a ubiquitous pollutant of the environment and a perilous material, its biodegradation has been receiving constant scientific consideration. Highly potential, naphthalene degrading bacteria were isolated from sediments of polluted Amlakadi canal, Gujarat, India. Among the isolates, Pseudomonas sp. HOB1, showed ability to degrade 2000 ppm naphthalene within 24h. The culture exhibited potential to tolerate as high as 60,000 ppm of naphthalene. Statistical approach was used to analyze the effect of physiological parameters and initial biomass concentration on naphthalene degradation. Naphthalene degradation was found to be augmented in the pH range of 7.5-8.5. Naphthalene degradation was maximum in the temperature range of 35-37 degrees C and initial inoculum size of more than 1.8 ml of 1.0 A(660). Simulated microcosm studies in the presence and absence of indigenous microflora confirmed its ability for naphthalene degradation and to colonize the soil. Pseudomonas sp. HOB1 was found to be highly potent in degrading higher concentrations of naphthalene under laboratory conditions as well as in simulated microcosms.

Activity of Trypsin Entrapped in Reverse Micelles

Journal of Surface Science and Technology, 1999

Trypsin (3.4.21.4) was entrapped in reverse micelles of AOT/isooctane. It was observed that the e... more Trypsin (3.4.21.4) was entrapped in reverse micelles of AOT/isooctane. It was observed that the enzyme remains active in reverse micelles. Activity was influencced by various factors like watertsurfactant ratio, pH, surfactant concentration. These parameters were studied in comparison with the conventional aqueous system and the results are reported.

Biomethanation of a mixture of salty cheese whey and poultry waste or cattle dung

Applied Biochemistry and Biotechnology, Aug 1, 1996

This paper describes the results of a study aimed at improving the efficiency of anaerobic digest... more This paper describes the results of a study aimed at improving the efficiency of anaerobic digestion of salty cheese whey in combination with poultry waste or cattle dung. Best results were obtained when salty cheese whey was mixed with poultry waste in the ratio of 7:3, or cattle dung in the ration of 1:1, both on dry weight basis giving maximum gas production of 1.2 L/L of digester/d with enriched methane content of 64% and 1.3 L/L of digester/d having methane content of 63% respectively. Various conditions such as temperature and retention time have been optimized for maximum process performance.

Continuous hydrogen evolution by an immobilized combined system of Phormidium valderianum, Halobacterium halobium and Escherichia coli in a packed bed reactor

International Journal of Hydrogen Energy, Aug 1, 1995

Abstract Long-term hydrogen evolution (11 days) was obtained when Phormidium valderianum was coup... more Abstract Long-term hydrogen evolution (11 days) was obtained when Phormidium valderianum was coupled with a mixture of halophilic Halobacterium halobium and salt tolerant Escherichia coli under a cyclic illumination on/off cycle of 18 h dark and 6 h light in an immobilized packed bed reactor. Various conditions such as flow rate, temperature and pH have been optimized for maximum continuous evolution.

Lipase from Solvent-Tolerant Pseudomonas sp. DMVR46 Strain Adsorb on Multiwalled Carbon Nanotubes: Application for Enzymatic Biotransformation in Organic Solvents

Applied Biochemistry and Biotechnology, Sep 2, 2015

Immobilization of biocatalysts onto particulate carriers has been widely explored for recycling o... more Immobilization of biocatalysts onto particulate carriers has been widely explored for recycling of biocatalyst. However, surface properties often affect the amount of biocatalysts immobilized, their bioactivity and stability, hampering their wide applications. The aim of this work was to elucidate the importance of nanoimmobilization system in organic synthesis. The surface of multiwalled carbon nanotubes (MWCNTs) was functionalized with a mixture of concentrated acids to create an interface for enzyme immobilization. Successful functionalization and enzyme immobilization was structurally evidenced by transmision electron microscopy analysis and Fourier-transform infrared spectroscopy analysis. Furthermore, immobilized enzyme was exploited for the synthesis of flavoured ester ethyl butyrate in the presence of n-heptane. Optimized conditions for enhanced ester synthesis was found to be 8.5 pH, 40 °C, 150 rpm, 0.15:0.2 M substrate molar ratio (ethanol/butyric acid) and n-heptane as reaction medium. Utmost 81 % of ester synthesis was obtained using immobilized lipase quite higher in comparison to that of free lipase. The activation energy indicated a lower energy requirement for immobilization of lipase on the surface of functionalized MWCNTs. In summary, immobilization of lipase on functionalized MWCNTs by simple adsorption method displayed excellent properties for enzyme stability and reusability, indicating its potential for application in organic synthesis.

Surfactants in anaerobic digestion of salty cheese whey using upflow fixed film reactor for improved biomethanation

Process Biochemistry, Feb 1, 1998

... 11. Ranade, DR & Gadre, RV, Microbial Aspects of Anaerobic Digejstion, Laboratory Manual,... more

Characterization of an intact phycoerythrin and its cleaved 14kDa functional subunit from marine cyanobacterium Phormidium sp. A27DM

Process Biochemistry, Sep 1, 2011

In this study, we describe a series of experiments presenting the biochemical evidence for the cl... more

ALLOPHYCOCYANIN FROM A LOCAL ISOLATE<i>GEITLERINEMA</i>SP. A28DM (CYANOBACTERIA): A SIMPLE AND EFFICIENT PURIFICATION PROCESS

Journal of Phycology, Apr 1, 2010

Phycocyanin is a major protein produced by cyanobacteria, but very few phycocyanin-producing stra... more Phycocyanin is a major protein produced by cyanobacteria, but very few phycocyanin-producing strains have been reported. In the present study, response surface methodology (RSM) involving a central com posite design for four factors was successfully employed to optimize medium components for increased production of phycocyanin from Phormidium ceylanicum. The production of phycocyanin and interactions between sodium nitrate, calcium chloride, trace metal mix and citric acid stock were investigated and modeled. Under optimized condition P. ceylanicum was able to give 2.3-fold increase in phycocyanin pro duction in comparison to commonly used BG 11 medium in 32 days. We have demonstrated the extrac tion, purification and characterization of C-phycocyanin using novel method based on filtration and single step chromatography. The protein was extracted by repeated freeze-thaw cycles and the crude extract was filtered and concentrated in stirred ultrafiltration cell [UFC). The UFC concentrate was then subjected to a single ion exchange chromatographic step. A purity ratio of 4.15 was achieved from a start ing value of 1.05. The recovery efficiency of C-phycocyanin from crude extract was 63.50%. The purity was checked by electrophoresis and UV-Vis spectroscopy.

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Candida rugosa lipase immobilized in Triton-X100 microemulsion based organogels (MBGs) for ester synthesis

Process Biochemistry, 2008

The esterification ability of the lipase VII from Candida rugosa was studied in the quaternary wa... more The esterification ability of the lipase VII from Candida rugosa was studied in the quaternary water-in-oil microemulsion Triton-X100/water/1-hexanol/n-hexane based organogels. The gelatin concentration exerted a high influence on both the MBG formation and the esterification rate exhibiting highest esterification at 6% (w/v). Nature of cosurfactants and oil exploited in making of the MBGs also affected the lipase-catalyzed esterification, showing highest

Ester synthesis by lipase immobilized on silica and microemulsion based organogels (MBGs)

Process Biochemistry, Feb 1, 2001

Candida rugosa lipase was immobilized in gelatin containing Aerosol-OT (AOT) microemulsion-based ... more Candida rugosa lipase was immobilized in gelatin containing Aerosol-OT (AOT) microemulsion-based organogels (MBGs) and on silica gel. The highest condensation activity on n-caprylic acid with ethanol was observed at R=60. Selective removal of water was achieved using 1 M AOT in isooctane dry reverse micelles for MBG with R=70. This pretreatment of lipase containing MBG resulted in the enhanced esterification.

Concurrent purification and antioxidant activity of phycobiliproteins from Lyngbya sp. A09DM: An antioxidant and anti-aging potential of phycoerythrin in Caenorhabditis elegans

Process Biochemistry, Oct 1, 2014

Abstract The present study probes into the purification of phycobiliproteins, and characterizatio... more Abstract The present study probes into the purification of phycobiliproteins, and characterization of their in vitro anti-oxidant activity. Moreover, the study also demonstrates the use of antioxidant virtue of phycoerythrin in moderating the phenomenon of aging in Caenorhabditis elegans. Phycoerythrin, phycocyanin and allophycocyanin were purified successfully from Lyngbya sp. A09DM by ammonium sulfate fractionation appended with Triton X-100 intercession. The success of protocol was examined by a series of biochemical characterization like SDS-PAGE, native-PAGE, UV–visible spectroscopy and fluorescence spectroscopy ensuring purity, integrity and functionality of purified phycoerythrin, phycocyanin and allophycocyanin. Purified phycobiliproteins were evaluated for antioxidant and metal ion chelating activity by various in vitro antioxidant assay systems. Results showed significant and dose-dependent antioxidant as well as metal chelating potential of all phycobiliproteins in decreasing order of phycoerythrin > phycocyanin > allophycocyanin. Expansion in lifespan and improvement in pharyngeal pumping of C. elegans were noticed upon pre-treatment with phycoerythrin (100 μg ml−1). Moreover, phycoerythrin mediated increase in worm survival under oxidative stress revealed that the life expansion effect of phycoerythrin on nematode is in part by an action of its antioxidant virtue. These results collectively added up evidence in favor of the ‘free-radical theory of aging’. The present report, for the first time, describes antioxidant potential of phycoerythrin and its use in extending life-span of C. elegans.

Role of Surfactants on the Activity of Acid Phosphatase Incorporated in Reverse Micelles

Biocatalysis and Biotransformation, 2000

The effect of water content, pH, and substrate concentration on the activity of acid phosphatase ... more The effect of water content, pH, and substrate concentration on the activity of acid phosphatase entrapped in a reversed micellar system of bis(2-ethylhexyt)sodium sulfosuccinate (AOT)/isooctane, Triton X-100/xylene and cetyltrimethylammonium bromide (CTAB)/Triton X-100 have been studied. Amongst the various reverse micellar systems, CTAB/Triton X-100 mixed system was found to be the best for acid phosphatase. At a very low water-surfactant ratio (Wo), the activity was found to be less than that of the conventional aqueous system. At Wo = 20, the activity resembled to that of the aqueous system. At Wo = 50, the activity was seen to be enhanced by 1.9-fold.

Sweet sorghum: a potential resource for bioenergy production

Elsevier eBooks, 2020

Abstract Lignocellulosic biomass is a sustainable renewable energy source, having the potential t... more Abstract Lignocellulosic biomass is a sustainable renewable energy source, having the potential to replace heat, power, fuel, materials, and chemicals that are derived from fossil-based material. The production of second-generation biofuels from lignin-derived biomass has significantly improved in the last decades, which offer both energy and environmental benefits. However, the destruction of polymeric framework of lignin into low molecular weight reactive moieties has proven challenging. Thus lower lignin and higher carbohydrate content crops have the advantage to produce a substantial quantity of biofuel. Among the various lignocellulosic feedstocks, sweet sorghum contains higher carbohydrate and energy content. Alternate bioenergy sources, namely, sugarcane and sugar beet, place an immense pressure on worldwide food markets, but the conflict between food versus fuel can be avoided by utilizing alternate substrate, that is, sweet sorghum. Although there are major obstacles in terms of yield and production of biofuel from sweet sorghum, with time it may possibly replace the huge share of the fossil fuels’ requirement to meet the rising energy demand. This chapter mainly focused on the production, distribution, and potential applications of sweet sorghum in production of biofuels and their value-added bioproducts, including pretreatment with the novel process to produce bioethanol and biogas.

Reversed micellar extraction of an extracellular acid phosphatase from fermentation broth

Process Biochemistry, Nov 1, 2000

Abstract The extraction of an extracellular acid phosphatase from fermentation broth was carried ... more Abstract The extraction of an extracellular acid phosphatase from fermentation broth was carried out, using a reversed micellar system in an organic solvent. Process parameters were optimized for maximum extraction using 200 mM bis(2-ethylhexyl)sodium sulphosuccinate (AOT) in isooctane at 25°C. The extraction of acid phosphatase was affected by pH, ionic strength, surfactant concentration and aqueous/organic volume ratio. At pH 8.0 a maximum forward enzyme activity transfer of 75% at 50 mM KCl was achieved. The maximum recovery of acid phosphatase of about 29% was obtained using a 3:1 aqueous/organic volume ratio for the forward transfer while on aqueous/organic volume ratio of 1:1 was optimal for the backward transfer.

Biosynthesis of Indigo Dye by Newly Isolated Naphthalene-Degrading Strain Pseudomonas sp. HOB1 and its Application in Dyeing Cotton Fabric

Applied Biochemistry and Biotechnology, May 14, 2009

Indigo is one of the oldest dyes manufactured chemically and is mostly used in textile, food, and... more Indigo is one of the oldest dyes manufactured chemically and is mostly used in textile, food, and pharmaceutical industries. However, owing to the environmental hazards posed by the chemical production, the present scenario in the field stipulates a biosynthesis alternative for indigo production. The present study describes an indigenously isolated naphthalene-degrading strain Pseudomonas sp. HOB1 producing a blue pigment when indole was added in the growth medium. This blue pigment was analyzed by high-pressure thin-layer chromatography and other spectroscopic techniques which revealed it to be the indigo dye. Pseudomonas sp. HOB1 showed ability to produce 246 mg indigo liter(-1) of the medium. The K (m) for the enzyme naphthalene dioxygenase which is involved in indigo formation is 0.3 mM, and V (max) was as high as 50 nmol min(-1) mg dry biomass(-1). The bacterial indigo dye was further successfully applied for dyeing cotton fabrics. The high indigo productivity of Pseudomonas sp. HOB1 using naphthalene as growth substrate and its applicability on cotton fabrics, therefore, stems the probability of using this culture for commercial indigo production.

Isolation, characterization and decolorization of textile dyes by a mixed bacterial consortium JW-2

Dyes and Pigments, 2007

... The consortium had the ability to decolorize nine different dyes amongst 10 tested. Potential... more

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