Subject(s)
Amino Acid Oxidoreductases/isolation & purification , Isoenzymes/isolation & purification , Trichoderma/enzymology , Amino Acid Oxidoreductases/chemistry , Amino Acid Oxidoreductases/drug effects , Catalysis , Chromatography, Ion Exchange , Isoelectric Focusing , Isoenzymes/chemistry , Isoenzymes/drug effects , Molecular WeightABSTRACT
Some physicochemical properties of L-lysine oxidase from two strains of Trichoderma were studied. Content of metals and cofactors (Se, Zn, Cu, Fe, Co, Mn, Mo), amino acid analysis, secondary structure were estimated. The enzyme molecule from Trichoderma sp was found to contain both FAD and PQQ cofactors.
Subject(s)
Amino Acid Oxidoreductases/chemistry , Trichoderma/enzymology , Amino Acids/analysis , Chromatography, Gel , Circular Dichroism , Flavin-Adenine Dinucleotide/chemistry , PQQ Cofactor , Quinolones/chemistry , Species Specificity , Spectrometry, FluorescenceABSTRACT
A new fungal strain, Trichoderma sp., discovered in Moscow, produces the antitumor enzyme, lysine-oxidase, which demonstrates an anti-invasive effect in vitro and anti-metastatic activity in vivo. Maximal inhibition of the in vitro invasion of MM1 clone cells was obtained when the tumor cells were pretreated with 2.5 mU/ml of lysine-oxidase; the pretreatment caused a 1.9-times reduction in cell growth and a 1.6-times reduction in the invasive capacity. We studied its anti-metastatic effect on the spreading Lewis lung carcinoma (3LL) in mice from which the primary tumor had been removed. The administration of the enzyme (50 U/kg, i.v.) significantly decreased not only the extent but the number of lung metastases, as compared with the untreated mice. In addition to that, the lysine-oxidase treatment considerably increases the life-span of mice from which the primary tumor had been removed (200 days after 3LL implantation, lysine-oxidase treatment caused surviving of 50% mice in experimental group).
Subject(s)
Amino Acid Oxidoreductases/therapeutic use , Neoplasm Invasiveness , Neoplasm Metastasis/prevention & control , Trichoderma/enzymology , Animals , Mice , Mice, Inbred BALB C , Neoplasm Transplantation , Neoplasms, Experimental/pathologyABSTRACT
A new procedure is developed for purification of the antitumoral enzyme L-lysyl-alpha-oxidase from Trichoderma sp. The procedure included two steps: hydrophobic chromatography on butyl sylochrome C-80 and chromatography using biospecific sorbent AH-Sepharose. The simplified procedure enabled to increase distinctly the specific enzymatic activity from 30 to 50 IU/mg in the preparation obtained with a good yield.
Subject(s)
Amino Acid Oxidoreductases/isolation & purification , Trichoderma/enzymology , Chromatography/methodsABSTRACT
The influence of a new antitumor enzyme L-lysine alpha-oxidase on Lewis lung carcinoma spreading was studied in mice in which primary tumor had been removed. The enzyme was found to significantly decrease the extent and number of lung metastases as compared to mice which hadn't received L-lysine alpha-oxidase. This was matched by recovery of alveolar macrophages functional activity, as assessed by adenosine deaminase and 5' nucleotidase levels in these cells. Moreover, antimetastatic and cytostatic effect was confirmed by the measuring of polyamine concentration in mice erythrocytes.
Subject(s)
Amino Acid Oxidoreductases/therapeutic use , Neoplasm Metastasis/prevention & control , 5'-Nucleotidase/analysis , Adenosine Deaminase/analysis , Animals , Chromatography , Erythrocytes/analysis , Lung Neoplasms/surgery , Macrophages/analysis , Mice , Mice, Inbred C57BL , Polyamines/blood , Pulmonary Alveoli/analysisABSTRACT
Evidences on the antileukemic effect of L-lysine alpha-oxidase from Trichoderma sp. are presented. It is inferred that enzyme needs detail studying as a potential chemotherapeutic drug both in experimental and clinical research.
Subject(s)
Amino Acid Oxidoreductases/therapeutic use , Leukemia L1210/drug therapy , Leukemia P388/drug therapy , Leukemia, Experimental/drug therapy , Mitosporic Fungi/enzymology , Trichoderma/enzymology , Amino Acid Oxidoreductases/administration & dosage , Animals , Drug Evaluation , Male , Mice , Mice, Inbred Strains , Time FactorsABSTRACT
Relatively rapid procedure was developed for isolation and purification of the antitumor enzyme L-lysine alpha-oxidase. The procedure involved three steps instead of six or four steps described previously, with a yield of 64.4%. The homogenous preparation was obtained as shown by gel electrophoresis and ultracentrifugation. The antitumor activity of the enzyme both in vivo and in vitro is under investigation.
Subject(s)
Amino Acid Oxidoreductases/isolation & purification , Mitosporic Fungi/enzymology , Trichoderma/enzymology , Chromatography, DEAE-Cellulose , L-Amino Acid Oxidase , Substrate SpecificityABSTRACT
L-lysine-alpha-oxidase, isolated from a strain of Trichoderma sp. exhibited enzymological properties suitable for a chemotherapeutic drug used in experimental oncology.
Subject(s)
Amino Acid Oxidoreductases/isolation & purification , Antineoplastic Agents , Mitosporic Fungi/enzymology , Trichoderma/enzymology , Amino Acid Oxidoreductases/pharmacology , Antineoplastic Agents/isolation & purification , Chromatography, Gel , Hydrogen-Ion Concentration , Isoelectric Focusing , Kinetics , Molecular Weight , Species Specificity , Substrate SpecificityABSTRACT
L-lysine-alpha-oxidase, a new fungal enzyme catalyzing oxidative L-lysine deamination, was shown to have an inhibitory effect on the in vitro synthesis of DNA, RNA and proteins in human carcinoma ovarian (CaOv) cells.
Subject(s)
Amino Acid Oxidoreductases/pharmacology , Antineoplastic Agents , Mitosporic Fungi/enzymology , Neoplasms/metabolism , Trichoderma/enzymology , Cell Line , DNA, Neoplasm/biosynthesis , DNA, Neoplasm/drug effects , Depression, Chemical , Humans , Neoplasm Proteins/biosynthesis , RNA, Neoplasm/biosynthesis , RNA, Neoplasm/drug effectsABSTRACT
L-lysine-alpha-oxidase, a new fungal enzyme catalyzing oxidative deamination of L-lysine, exerts an inhibitory effect on DNA, RNA and protein synthesis in human cells of carcinoma ovarius (CaOv) in vitro.
Subject(s)
Amino Acid Oxidoreductases/pharmacology , DNA, Neoplasm/biosynthesis , Mitosporic Fungi/enzymology , RNA, Neoplasm/biosynthesis , Trichoderma/enzymology , Cells, Cultured , Depression, Chemical , Female , Humans , Neoplasm Proteins/biosynthesis , Ovarian Neoplasms/metabolismABSTRACT
An improved and relatively rapid procedure is developed for isolation and purification of a new antitumor enzyme L-lysyl-alpha-oxidase from Trichoderma sp. The method involves four steps, instead of six steps described previously, with a yield of 22.4%. The purified enzyme preparation was homogeneous as shown by polyacrylamide gel disc electrophoresis and ultracentrifugation. Physico-chemical and antitumor properties of the enzyme are under study.
Subject(s)
Amino Acid Oxidoreductases/isolation & purification , Mitosporic Fungi/enzymology , Trichoderma/enzymology , Chromatography, DEAE-Cellulose , Chromatography, Gel , Chromatography, Ion Exchange , Electrophoresis, DiscABSTRACT
L-lysine-alpha-oxidase from Trichoderma sp. fungi inhibits the synthesis of DNA, RNA and protein in cultivated cells of the Burkitt lymphoma (strain P3HRj) in vitro, which results in the blocking of the cells' transfer from phase S to phase G2/M of the mitotic cycle.
Subject(s)
Amino Acid Oxidoreductases/toxicity , Burkitt Lymphoma/pathology , Burkitt Lymphoma/metabolism , Cell Cycle/drug effects , Cells, Cultured , DNA, Neoplasm/biosynthesis , Dose-Response Relationship, Drug , Drug Evaluation, Preclinical , Humans , Neoplasm Proteins/biosynthesis , RNA, Neoplasm/biosynthesis , Time Factors , Trichoderma/enzymologyABSTRACT
Trichoderma cultures were tested for their ability to produce L-lysine-alpha-oxidase. The highest enzyme activity was manifested by T. harzianum (MGU), T. longibrachiatum Rifai VKM F-2025 and T. aureoviride Rifai VKM F-2026. The biosynthesis of the enzyme did not depend on the growth of the cultures and did not vary among the species.