A low molecular mass alkaliphilic extra-cellular lipase of Bacillus cereus MTCC 8372 was purified 35-fold by hydrophobic interaction (Octyl-Sepharose) chromatography. The purified enzyme was found to be electrophoretically pure by denaturing gel electrophoresis and possessed a molecular mass of approximately 8 kDa. It is a homopentamer of 40 kDa as revealed by native-PAGE. The lipase was optimally active at 55 °C and retained approximately half of its original activity after 40 min incubation at 55 °C. The enzyme was maximally active at pH 8.5. Mg2+, Cu2+, Ca2+, Hg2+, Al3+ and Fe3+ at 1 mM enhanced hydrolytic activity of the lipase. Interestingly, Hg2+ ions synergized and Zn2+ and Co2+ ions antagonized the lipase activity. Among surfactants, Tween 80 promoted the lipase activity. Phenyl methyl sulfonyl fluoride (PMSF, 15 mM) decreased 98% of original activity of lipase. The lipase was highly specific towards p-nitrophenyl palmitate and showed a Vmax and Km of 0.70 mmol.mg−1.min−1 and 32 mM for hydrolysis of pNPP.
Authors:R. Verma, L. Verma, M. Ranjan, B. Verma and S. Mojumdar
A series of ternary complexes of the types M2L′2L″2;ML′2L″2 (M=Fe, Cu, Zn; L′=2-oxocyclopentane dithiocarboxylate; L″=pyridine, morpholine) and CuL′2H2O was prepared afresh. Except the iron complex, all are dimer and complexation is through the dithio moiety of the ligand
L′. Their thermal decomposition was carried out in air at heating rate 10°C min−1 and it revealed that the dehydration of the aqua complex follows the same path as the carboxylates and the pyridine complexes
have the tendency to follow one-step decomposition. The copper complexes are less thermally stable. The overall thermal stability
of the 2-oxocyclopentanedithiocarboxylato complexes of the three metals with the volatile ligands was found to be in the order:
(CuLmorph)2< CuL2H2O<(CuLpy)2<(ZnLmorph)2<(ZnLpy)2<FeL2py2. The thermogravimetric properties of the complexes have been studied and the data were subjected to kinetic analysis. The
values of n, E, A and ΔS# have been approximated and compared. Any formation of bridged structure is not indicated in the first step case.
Authors:R. K. Verma, L. Verma, M. Chandra and A. Bhushan
Summary A comparative study of the non-isothermal decomposition of the dl-lactate hydrates of magnesium, calcium and strontium has been made with that of the dl-lactate hydrates chromium(III), manganese(II), iron(II), cobalt(II), nickel(II), copper(II) and zinc(II) keeping dry air as the purge gas and the heating rate maintained at 10 K min-1. While the dl-lactates of manganese(II), cobalt(II) and copper(II) followed single step decomposition scheme suggesting that dehydration and decomposition steps overlapped, the dehydration steps of the other compounds were distinct. &-T plots of none of the dehydration steps showed any induction period, indicating no physical desorption, nucleation or branching. Neither the & max-values nor the onset temperatures of the dehydration steps did show any pattern. The TG data of the dehydration steps have also been analyzed using the Freeman-Carroll, Horowitz-Metzger, Coats-Redfern, Zsakó, Fuoss-Salyer-Wilson and Karkhanavala-Dharwadkar methods. Values of order of reaction, activation energy and Arrhenius factor have been approximated and compared. There are similarities in the activation energy values for the dehydration steps (< 60 kJ mol-1 in general). It is higher with group 2 metals and lower in transition metals (maximum in magnesium and lowest in chromium and iron lactates). In cases of overlapping of dehydration and decomposition steps, the activation energy values are on the lower side with the same trend (lower in cobalt and copper cases).
In recent times, biotechnological applications of microbial lipases in synthesis of many organic molecules have rapidly increased in non-aqueous media. Microbial lipases are the ‘working horses’ in biocatalysis and have been extensively studied when their exceptionally high stability in non-aqueous media has been discovered. Stability of lipases in organic solvents makes them commercially feasibile in the enzymatic esterification reactions. Their stability is affected by temperature, reaction medium, water concentration and by the biocatalyst’s preparation. An optimization process for ester synthesis from pilot scale to industrial scale in the reaction medium is discussed. The water released during the esterification process can be controlled over a wide range and has a profound effect on the activity of the lipases. Approaches to lipase catalysis like protein engineering, directed evolution and metagenome approach were studied. This review reports the recent development in the field of non-aqueous microbial lipase catalysis and factors controlling the esterification/transesterification processes in organic media.
A purified alkaline thermo-tolerant bacterial lipase from
MTCC 8372 was immobilized on a
-MBAm) hydrogel. The hydrogel showed approximately 94% binding capacity for lipase. The immobilized lipase (2.36 IU) was used to achieve esterification of myristic acid and isopropanol in
-heptane at 65 °C under continuous shaking. The myristic acid and isopropanol when used at a concentration of 100 mM each in
-heptane resulted in formation of isopropyl myristate (66.0 ± 0.3 mM) in 15 h. The reaction temperature below or higher than 65°C markedly reduced the formation of isopropyl myristate. Addition of a molecular sieve (3 Å × 1.5 mm) to the reaction mixture drastically reduced the ester formation. The hydrogel bound lipase when repetitively used to perform esterification under optimized conditions resulted in 38.0 ± 0.2 mM isopropyl myristate after the 3
cycle of esterification.
A wide range of fatty acid esters can be synthesized by esterification and transesterification reactions catalyzed by lipases in non-aqueous systems. In the present study, immobilization of a purified alkaline extra-cellular lipase of
MTCC 8372 by adsorption on diatomaceous earth (celite) for synthesis of ethyl acetate via transesterification route was investigated.
lipase was deposited on celite (77% protein binding efficiency) by direct binding from aqueous solution. Immobilized lipase was used to synthesis of ethyl acetate from vinyl acetate and ethanol in
-nonane. Various reaction conditions, such as biocatalyst concentration, substrates concentration, choices of solvents (
-alkanes), incubation time, temperature, molecular sieves (3Å × 1.5 mm), and water activity(a
), were optimized. The immobilized lipase (25 mg/ml) was used to perform transesterification in
-alkane(s) that resulted in approximately 73.7 mM of ethyl acetate at 55 °C in
-nonane under shaking (160 rpm) after 15 h, when vinyl acetate and ethanol were used in a equimolar ratio (100 mM each). Addition of molecular sieves (3Å × 1.5 mm) as well as effect of water activity of saturated salt solutions (KI, KCl and KNO
) to the transesterification efficiency has inhibitory effect. Batch operational stability tests indicated that immobilized lipase had retained 50% of its original catalytic activity after four consecutive batches of 15 h each.
The reinforcement of nano-barium titanate in ferrite filled poly-ether-ether-ketone (PEEK) composites caused a shift in the
decomposition temperature, at which maximum mass loss occurred, to higher side and enhancement in char yield in thermogravimetric
analysis. Loss tangent and glass transition temperature of ferrite filled PEEK composites were also found to be increased
with the reinforcement of nano barium titanate. The effect of nano barium titanate on the melting behaviour of ferrite filled
PEEK composites was negligible.
The glass-forming tendency and specific heat in ice cold water-quenched Ge1−xSnxSe2.5 glassy alloys with 0<x<0.6 were investigated by means of differential scanning calorimetry. The heat of fusion ΔHf, the heat ΔHc associated with the crystallization of an amorphous phase and the glass transition temperatureTg were deduced from the DSC curves. The composition dependence of glass forming ability,Tg and crystallization behavior has been discussed.
Authors:Deepak Verma, M. Shakir Khan and Mohd. Zubair
Effective radium content and radon exhalation rates in soil samples have been measured by “Sealed Can Technique” using LR-115
type II plastic track detectors. The soil samples were collected from Farrukhabad city of Utter Pradesh, India. The values
of effective radium content were found to vary from 5.39 to 34.56 Bq kg−1 with an average value of 16.58 Bq kg−1 and a standard deviation of 7.16. The mass and surface exhalation rate has been found to vary from 0.41 × 10−6 to 2.64 × 10−6 Bq kg−1 d−1 and 1.41 × 10−6 to 9.10 × 10−6 Bq m−2 d−1, respectively. All the values of radium content in soil samples of study area were found to be quite lower than the permissible
value of 370 Bq kg−1 recommended by Organization for Economic Cooperation and Development.
Authors:B. M. Gupta, Avinash Kshitij and Charu Verma
The research output of India in computer science during 1999–2008 is analyzed in this paper on several parameters including total research output, its growth, rank and global publication share, citation impact, share of international collaborative papers and major collaborative partner countries and patterns of research communication in most productive journals. It also analyses the characteristics of most productive institutions, authors and high-cited papers. The publications output and impact of India is also compared with China, South Korea, Taiwan and Brazil.