Phytochemicals and Mineral Potentials of Tamarindus Indica L Seeds as Influenced by Processing Treatments

Tamarindus belongs to the subfamily Caesalpinioideae. Tamarindus itself is a monotypic genus, containing the sole species T. indica. Tamarind is a multi-use tree. The present study was outlined to investigate the phytochemicals and mineral composition of Tamarindus indica L seeds as influenced by processing treatments such as soaking,dehulling, cooking, autoclaving and germinating the seeds. Extraction of Tamarindus indica L seeds were carried out at 37°C with methanol and the extracted materials were then determined for the presence of phytochemicals. The alkaloids, steroids, flavonoids,terpenoids, tannins and phenolics were present in the soaked, cooked and autoclaved samples. Absence of phytochemicals was observed in case of dehulled seeds. But in case of germinated seeds, the presence of alkaloids, flavonoids and phenolics was noted. Further the ash content of the sample was determined and the ash obtained from all the processed samples were extracted with concentrated HCl and was subjected to mineral analysis. In soaked seeds, potassium and sodium decreased when compared to the control unprocessed seeds. In contrast sodium increased in dehulled and autoclaved seeds.Calcium and magnesium remains same in the entire processed sample as that of control group except that of dehulled seeds where magnesium is decreased and calcium is increased. Iron and phosphorus also remains similar as that of the control group except in case of germinated seeds where there is an increase in their content

Studies on Xyloglucanase during the Germination of Seeds of Tamarindus Indica

Germinating seeds of Tamarindus indicacontain endo-β-1, 4-xyloglucanases which degrade tamarind xyloglucan, but notcarboxymethylcellulose (CMC). The xyloglucanases are isolated from thegerminating tamarind seeds using 50 mM acetate buffer, pH 5.5 containing 0.5 MNaCl. The Km value is 0.667 g/liter and the enzyme is optimally active at pH5.5 and stable between pH 4 - 6.5. The optimum temperature is 45C and is quitestable upto 50C. The activity declined by 50% at 60C and is completelyinactivated at 70C. Highest xyloglucanase activity and specific activity areobserved on the 23rd day of germination. The polyacrylamide gel electrophoresis(PAGE) indicated the presence of five isozymes of xyloglucanases which arevisualized by activity staining separately with congo red and grams iodine.Isozyme 2 is the major xyloglucanase present throughout the germination period

Purification, characterization and properties of carboxylesterase from the midgut of the silkworm, Bombyx mori L.

A carboxylesterase has been purified from the midgut of the silkworm Bombyx mon L. by a combination of ammonium sulphate fractionation, DEAE-cellulose ion-exchange chromatography, Sephacryl S-200 gel-filtration and preparative polyacrylamide gel electrophoresis (PAGE). The homogeneity of the enzyme was established by PAGE, isoelectricfocusing (IEF) and SDS-PAGE. The enzyme consists of two identical subunits with a subunit molecular weight of 72,000. The two subunits are held by non-covalent bonds. Amino acid analysis of the purified enzyme revealed a high content of hydrophobic amino acid residues. It lacks proline and tryptophan residues and free thiol groups. The data from substrate specificity study in conjunction with kinetic parameters indicate the hydrophobic nature of the active site. Copyright © 1996 Elsevier Science Ltd. All rights reserved

The Nobel Prize in Chemistry 2004: 'ubiquitous' quality control of life

The Nobel Prize in Chem. for 2004 is shared by Aaron Ciechanover, Avram Hershko and Irwin Rose, who made fundamental discoveries concerning how cells regulate the breakdown of cellular proteins with extreme specificity. The three biochemists discovered ubiquitin-​mediated proteolysis, a process where an enzyme system tags unwanted proteins with many mols. of a small protein called ubiquitin and then sends then to the proteasome where they are broken down

Characterization of Esterases of Tamarindus Indica Seeds

Germinating seeds of Tamarindus indicasynthesizes various enzymes which are required for the degradation of seedreserves such as xyloglucans, fatty acid esters and proteins. Among these,esterases, belonging to a group of hydrolytic enzymes catalyze the hydrolysisof various types of esters. They play an important role in cell expansion aswell as detoxification of xenobiotics and many agrochemicals and insecticides.The esterases are extracted from the germinating tamarind seeds using 50 mMphosphate buffer, pH 7. The Km with α-naphthyl acetate as the substrate is19.23 μM and the enzymes are optimally active at pH 7.0 to 7.5 and are stablebetween pH 5.0 to 9.0. The optimum temperature of esterase activity of tamarindseed is between 37C - 50C and is stable up to 40C. The activity declined by30% at 60C and about 90% at 70C. Highest esterase activity and specificactivity are observed on the 21st day of germination. The polyacrylamide gelelectrophoresis (PAGE) indicated the presence of nine isozymes of esterases.Band numbers 1, 5 and 6 are the major esterolytic bands present throughout thegermination period while band numbers 2 & 3 are minor bands present onlyduring the latter period of the germination. Based on substrate and inhibitorspecificity in conjunction with electrophoresis, the esterases 1 to 8 have beenclassified as carboxylesterases sensitive to organophosphate inhibitor (OP) andPCMB (p-chloromercuribenzoate) while esterase 9 is classified as carboxylesterasesensitive to OP. These esterases are unaffected by carbamate inhibitor, eserinesulphate

Carboxylesterases from the seeds of an underutilized legume, Mucuna pruriens; Isolation, purification and characterization

Two carboxylesterases (ME-III and ME-IV) have been purified to apparent homogeneity from the seeds of Mucuna pruriens employing ammonium sulfate fractionation, cation exchange chromatography on CM-cellulose, gel-permeation chromatography on Sephadex G-100 and preparative PAGE. The homogeneity of the purified preparations was confirmed by polyacrylamide gel electrophoresis (PAGE), gel-electrofocussing and SDS-PAGE. The molecular weights determined by gel-permeation chromatography on Sephadex G-200 were 20.89 kDa (ME-III) and 31.62 kDa (ME-IV). The molecular weights determined by SDS-PAGE both in the presence and absence of 2-mercaptoethanol were 21 kDa (ME-III) and 30.2 kDa (ME-IV) respectively, suggesting a monomeric structure for both the enzymes. The enzymes were found to have Stokes radius of 2.4 nm (ME-III) and 2.7 nm (ME-IV). The isoelectric pH values of the enzymes, ME-III and ME-IV, were 6.8 and 7.4, respectively. ME-III and ME-IV were classified as carboxylesterases employing PAGE in conjunction with substrate and inhibitor specificity. The K m of ME-III and ME-IV with 1-naphthyl acetate as substrate was 0.1 and 0.166 mM while with 1-naphthyl propionate as substrate the K m was 0.052 and 0.0454 mM, respectively. As the carbon chain length of the acyl group increased, the affinity of the substrate to the enzyme increased indicating hydrophobic nature of the acyl group binding site. The enzymes exhibited an optimum temperature of 45 °C (ME-III) and 37 °C (ME-IV), an optimum pH of 7.0 (ME-III) and 7.5 (ME-IV) and both the enzymes (ME-III and ME-IV) were stable up to 120 min at 35 °C. Both the enzymes were inhibited by organophosphates (dichlorvos and phosphamidon), but resistant towards carbamates (carbaryl and eserine sulfate) and sulphydryl inhibitors (p-chloromercuricbenzoate, PCMB). © 2011 Elsevier Ltd. All rights reserved

CHARACTERIZATION OF ALPHA-AMYLASE FROM THE SEEDS OF Mucuna pruriens

Amylases are hydrolytic enzymes which are widely distributed in nature, animals, plants and microorganisms. Amylases are of great significance in present-day biotechnology. In present study, amylases are isolated from the soaked seeds of Mucuna pruriens under extreme acidic conditions. Conventional protein purification techniques such as salt fractionation, ion exchange chromatography on CM-cellulose and sephadex G-75 was employed for the purification of amylase from the seeds of Mucuna pruriens. The amylase activity was eluted in one peak. The specific activity and yield of the purified amylase was 6.25 and 29.99, respectively. Native PAGE, SDS-PAGE and gel electrofocussing were employed to establish homogeneity of the purified amylase. SDS-PAGE and gel-filtration chromatography on sephadex G-75 was used to determine the molecular weight of the purified amylase. The purified amylase was nearly homogenous and its molecular weight was found to be 78.4 kDa. The optimum pH and temperature of the purified amylase were 7.0 and 50oC, respectively. The isolectric pH of the purified amylase was 7.2 and the activity was linear up to 60 minutes

Esterase Activity from the Germinated Jatropha curcas Seeds in Different Extraction buffers.

The buffer solution plays a major role in protein stability and activity, thereby making the selection of a buffer to achievemaximum activity for a protein will be a formidable challenge. The present work constitutes an extension of this investigation to esterases from germinated Jatrophacurcas seeds. The 0.1 M NaCl solution, 0.1 M phosphate buffer pH7.0, 0.1 M citrate buffer pH 5.0 and 0.1 M Tris-HCl buffer pH 8.5, 0.1 M NaOH and distill water were used to extract protein from germinated Jatrophacurcas seeds. The esterase activity and specific activity for NaCl solution, phosphatebuffer, citrate buffer, Tris-HCl buffer, NaOH and Distilled water was 9.07, 8.6, 8.2, 6.46, 0.07 and 4.98 μmoles/min/gm and 0.09258, 0.0905, 0.088, 0.0715 0.0003 and 0.081 IU/mg, respectively.The Native-PAGE analysis showed the esterase enzyme activity in different extraction buffer.Among 13 esterase bands, 8 esterolytic bands were major bands (band no 1,3, 6, 7, 8, 11, 12 and 13)and remaining were minor bands.The amount of proteins and esterase activity were found to bethe highest when extracted with 0.1 M NaCl solution