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Braz. j. microbiol ; Braz. j. microbiol;46(4): 957-968, Oct.-Dec. 2015. tab, graf
Article in English | LILACS | ID: lil-769664

ABSTRACT

Abstract L-glutaminase was produced by Streptomyces canarius FR (KC460654) with an apparent molecular mass of 44 kDa. It has 17.9 purification fold with a final specific activity 132.2 U/mg proteins and 28% yield recovery. The purified L-glutaminase showed a maximal activity against L-glutamine when incubated at pH 8.0 at 40 °C for 30 min. It maintained its stability at wide range of pH from 5.0 11.0 and thermal stable up to 60 °C with Tm value 57.5 °C. It has high affinity and catalytic activity for L-glutamine (Km 0.129 mM, Vmax 2.02 U/mg/min), followed by L-asparagine and L-aspartic acid. In vivo, L-glutaminase showed no observed changes in liver; kidney functions; hematological parameters and slight effect on RBCs and level of platelets after 10 days of rabbit's injection. The anticancer activity of L-glutaminase was also tested against five types of human cancer cell lines using MTT assay in vitro. L-glutaminase has a significant efficiency against Hep-G2 cell (IC50, 6.8 μg/mL) and HeLa cells (IC50, 8.3 μg/mL), while the growth of MCF-7 cells was not affected. L-glutaminase has a moderate cytotoxic effect against HCT-116 cell (IC50, 64.7 μg/mL) and RAW 264.7 cell (IC50, 59.3 μg/mL).


Subject(s)
Animals/chemistry , Animals/drug effects , Animals/enzymology , Animals/metabolism , Animals/pharmacology , Antineoplastic Agents/chemistry , Antineoplastic Agents/drug effects , Antineoplastic Agents/enzymology , Antineoplastic Agents/metabolism , Antineoplastic Agents/pharmacology , Biocatalysis/chemistry , Biocatalysis/drug effects , Biocatalysis/enzymology , Biocatalysis/metabolism , Biocatalysis/pharmacology , Cell Proliferation/chemistry , Cell Proliferation/drug effects , Cell Proliferation/enzymology , Cell Proliferation/metabolism , Cell Proliferation/pharmacology , Enzyme Stability/chemistry , Enzyme Stability/drug effects , Enzyme Stability/enzymology , Enzyme Stability/metabolism , Enzyme Stability/pharmacology , Glutaminase/chemistry , Glutaminase/drug effects , Glutaminase/enzymology , Glutaminase/metabolism , Glutaminase/pharmacology , Glutamine/chemistry , Glutamine/drug effects , Glutamine/enzymology , Glutamine/metabolism , Glutamine/pharmacology , HeLa Cells/chemistry , HeLa Cells/drug effects , HeLa Cells/enzymology , HeLa Cells/metabolism , HeLa Cells/pharmacology , /chemistry , /drug effects , /enzymology , /metabolism , /pharmacology , Humans/chemistry , Humans/drug effects , Humans/enzymology , Humans/metabolism , Humans/pharmacology , Kinetics/chemistry , Kinetics/drug effects , Kinetics/enzymology , Kinetics/metabolism , Kinetics/pharmacology , Streptomyces/chemistry , Streptomyces/drug effects , Streptomyces/enzymology , Streptomyces/metabolism , Streptomyces/pharmacology , Substrate Specificity/chemistry , Substrate Specificity/drug effects , Substrate Specificity/enzymology , Substrate Specificity/metabolism , Substrate Specificity/pharmacology
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