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A surface membrane 3'-nucleotidase from Leishmania donovani promastigotes has been purified to SDS/PAGE homogeneity. The enzyme has apparent subunit molecular mass of 38 kDa, pI 5.8 and a broad pH optimum, 5.5-7.5. EDTA partially inhibited the enzyme activity, which was fully restored by Co2+; Mg2+, Ca2+ or Mn2+ had no effect on the activity. ZnCl2 or dithiothreitol at 1 mM was inhibitory at pH...
The alpha-amylases in the salivary glands of Lygus hesperus Knight and L. lineolaris (Palisot de Beauvois) were isolated and purified by ion exchange chromatography, and by isoelectric focusing, respectively. The alpha-amylase from L. hesperus had an isoelectric point (pI) of 6.25, and a pH optimum of 6.5. The specific activity of alpha-amylases in the salivary glands of L. hesperus was 1.2 U/m...
Bacillus pumilus 2.g isolated from gembus, an Indonesian fermented soybean cake, secretes several proteases that have strong fibrinolytic activities. A fibrinolytic enzyme with an apparent molecular weight of 20 kDa was purified from the culture supernatant of B. pumilus 2.g by sequential application of ammonium sulfate precipitation, ion-exchange chromatography, and hydrophobic chromatography....
The characterization of a spore laccase from Bacillus vallismortis fmb-103, isolated from textile industry disposal sites, is described. The activity was 6.5 U/g of dry spore with ABTS as the substrate. The enzyme was quite stable at high temperature. It retained more than 90% of its initial activity after 10h at 70 °C. The enzyme demonstrated broad pH stability in both acidic and alkaline cond...
Acinetobacter sp. isolated from healthy human skin of a tribal population was tested for lipase production.Medium optimization was achieved to increase the production. Purification was carried out by a one-step purification process using DEAE Sephadex A-50. The molecular weight of the lipase was approximately 60 kDa by SDS-PAGE. The purified lipase showed not only good stability in the presence...
Mouse C1q, a subcomponent of the first component of complement, was purified from mouse EDTA plasma by a combination of precipitation with polyethyleneglycol, affinity chromatography on IgG-Sepharose, ion exchange chromatography and molecular sieving. Mouse C1q was compatible with human C1q in the sense that it shows C1 hemolytic activity by the combination with human C1-r and C1-s. The molecul...
In this study, we used three different methods for the extraction of membrane-bound bilirubin (EMB) from erythrocyte membranes. Use of 2.5% albumin, pH 7.4, for elution of EMB resulted in only 34% of the total EMB which was estimated after the solubilization of bilirubin-loaded erythrocyte membranes (BLEMs) with 1% SDS. On the other hand, incubation of BLEMs with 38 mM sodium carbonate solution...
An intracellular oligopeptidase from Lactobacillus paracasei Lc-01 has been purified to homogeneity by Fast Flow Q Sepharose, hydroxyapatite, and Mono Q chromatography. The molecular mass of the enzyme was determined to be 140 kDa by gel filtration and approximately 30 kDa by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis and SDS-capillary electrophoresis. The pI of the enzyme ...
A gene (gkdA) (741 bp) encoding a putative protein of 247 amino acids was cloned from the Bacillus licheniformis SR01. The protein was expressed in Escherichia coli BL21 with a molecular mass estimated by SDS-PAGE of approximately 28.03 kDa and showed a calculating isoelectric point (pI) of 6.42. Structure analysis and function identification showed that the enzyme was a multifunctional glycosi...
Bacterial L-asparaginase has been widely used as therapeutic agent in treatment of various lymphoblastic leukemia and food processing aid to reduce the formation of cancer causing acrylamide. The present work deals with production and purification of extracellular L-asparaginase from soil isolates using solid state fermentation. The isolate was characterized by big chemical test and identified ...
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