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/pharmacologyABSTRACT
Chloramphenicol and kinetin stimulated the production of alkaloids and ascorbic acid in young Celosia plants. Different concentrations of chloramphenicol and kinetin were applied as foliar spray on four weeks old plants. Chloramphenicol [120 mg/l] and kinetin [30 mcg/1] showed beneficial effect on total alkaloid and ascorbic acid production in young plants. The total alkaloid was determined by precipitation with ammonium reineckate; while the ascorbic acid was estimated colorimetrically. The total alkaloid had increased to 0.147% in root powder as compared with control [0.078%] and had also increased in stem and leaves. The better yield of ascorbic acid [0.587%] showed in leaves as compared with control [0.396%]