Assuntos
Aminoácido Oxirredutases/isolamento & purificação , Isoenzimas/isolamento & purificação , Trichoderma/enzimologia , Aminoácido Oxirredutases/química , Aminoácido Oxirredutases/efeitos dos fármacos , Catálise , Cromatografia por Troca Iônica , Focalização Isoelétrica , Isoenzimas/química , Isoenzimas/efeitos dos fármacos , Peso MolecularRESUMO
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.
Assuntos
Aminoácido Oxirredutases/química , Trichoderma/enzimologia , Aminoácidos/análise , Cromatografia em Gel , Dicroísmo Circular , Flavina-Adenina Dinucleotídeo/química , Cofator PQQ , Quinolonas/química , Especificidade da Espécie , Espectrometria de FluorescênciaRESUMO
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).
Assuntos
Aminoácido Oxirredutases/uso terapêutico , Invasividade Neoplásica , Metástase Neoplásica/prevenção & controle , Trichoderma/enzimologia , Animais , Camundongos , Camundongos Endogâmicos BALB C , Transplante de Neoplasias , Neoplasias Experimentais/patologiaRESUMO
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.
Assuntos
Aminoácido Oxirredutases/isolamento & purificação , Trichoderma/enzimologia , Cromatografia/métodosRESUMO
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.
Assuntos
Aminoácido Oxirredutases/uso terapêutico , Metástase Neoplásica/prevenção & controle , 5'-Nucleotidase/análise , Adenosina Desaminase/análise , Animais , Cromatografia , Eritrócitos/análise , Neoplasias Pulmonares/cirurgia , Macrófagos/análise , Camundongos , Camundongos Endogâmicos C57BL , Poliaminas/sangue , Alvéolos Pulmonares/análiseRESUMO
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.
Assuntos
Aminoácido Oxirredutases/uso terapêutico , Leucemia L1210/tratamento farmacológico , Leucemia P388/tratamento farmacológico , Leucemia Experimental/tratamento farmacológico , Fungos Mitospóricos/enzimologia , Trichoderma/enzimologia , Aminoácido Oxirredutases/administração & dosagem , Animais , Avaliação de Medicamentos , Masculino , Camundongos , Camundongos Endogâmicos , Fatores de TempoRESUMO
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.
Assuntos
Aminoácido Oxirredutases/isolamento & purificação , Fungos Mitospóricos/enzimologia , Trichoderma/enzimologia , Cromatografia DEAE-Celulose , L-Aminoácido Oxidase , Especificidade por SubstratoRESUMO
L-lysine-alpha-oxidase, isolated from a strain of Trichoderma sp. exhibited enzymological properties suitable for a chemotherapeutic drug used in experimental oncology.
Assuntos
Aminoácido Oxirredutases/isolamento & purificação , Antineoplásicos , Fungos Mitospóricos/enzimologia , Trichoderma/enzimologia , Aminoácido Oxirredutases/farmacologia , Antineoplásicos/isolamento & purificação , Cromatografia em Gel , Concentração de Íons de Hidrogênio , Focalização Isoelétrica , Cinética , Peso Molecular , Especificidade da Espécie , Especificidade por SubstratoRESUMO
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.
Assuntos
Aminoácido Oxirredutases/farmacologia , Antineoplásicos , Fungos Mitospóricos/enzimologia , Neoplasias/metabolismo , Trichoderma/enzimologia , Linhagem Celular , DNA de Neoplasias/biossíntese , DNA de Neoplasias/efeitos dos fármacos , Depressão Química , Humanos , Proteínas de Neoplasias/biossíntese , RNA Neoplásico/biossíntese , RNA Neoplásico/efeitos dos fármacosRESUMO
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.
Assuntos
Aminoácido Oxirredutases/farmacologia , DNA de Neoplasias/biossíntese , Fungos Mitospóricos/enzimologia , RNA Neoplásico/biossíntese , Trichoderma/enzimologia , Células Cultivadas , Depressão Química , Feminino , Humanos , Proteínas de Neoplasias/biossíntese , Neoplasias Ovarianas/metabolismoRESUMO
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.
Assuntos
Aminoácido Oxirredutases/isolamento & purificação , Fungos Mitospóricos/enzimologia , Trichoderma/enzimologia , Cromatografia DEAE-Celulose , Cromatografia em Gel , Cromatografia por Troca Iônica , Eletroforese DescontínuaRESUMO
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.
Assuntos
Aminoácido Oxirredutases/toxicidade , Linfoma de Burkitt/patologia , Linfoma de Burkitt/metabolismo , Ciclo Celular/efeitos dos fármacos , Células Cultivadas , DNA de Neoplasias/biossíntese , Relação Dose-Resposta a Droga , Avaliação Pré-Clínica de Medicamentos , Humanos , Proteínas de Neoplasias/biossíntese , RNA Neoplásico/biossíntese , Fatores de Tempo , Trichoderma/enzimologiaRESUMO
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.