细胞色素p450系统可能与高等真菌的发光有关

N. Ronzhin, E. Posokhina, O. Mogilnaya, A. Puzyr, J. Gitelson, V. Bondar
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引用次数: 0

摘要

本文提出的数据证明,有利于细胞色素P450系统在高等真菌的发光参与。从不同种类的发光担子菌的菌丝体中提取含有真菌发光系统的提取物,这些真菌发光系统在体外提供发光。发光系统分离的应用条件(超声、40000g离心)表明,提取物中存在膜结构,特别是内质网(ER)超声解体形成的微粒体。在410 nm和450 nm处存在两个吸收峰,表明存在细胞色素b5和P450。提取物的发光是由还原的吡啶核苷酸刺激的,然而,NADPH的加入比NADH引起更高的发光水平。过氧化氢的加入显著提高了NAD(P)H活化提取物的发光强度(从几倍到1-2个数量级)。氟康唑的加入显著抑制了提取物的发光。结果表明,与ER膜相关的细胞色素P450系统可能参与了高等真菌的发光机制,参与了nadph依赖的细胞色素P450还原酶-细胞色素P450和nadh依赖的细胞色素b5还原酶-细胞色素b5 -细胞色素P450的电子传递酶系统过程。在这种情况下,细胞色素P450可能羟化hispidin(发光反应底物的前体)形成荧光素,并在ROS存在的情况下通过发光催化其氧化。
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CYTOCHROME P450 SYSTEM MAY BE INVOLVED IN THE LIGHT EMISSION OF HIGHER FUNGI
The paper presents data that testify in favor of the participation of the cytochrome P450 system in the light emission of higher fungi. Extracts from mycelia of different species of luminous basidiomycetes containing fungal luminescent systems that provide luminescence in vitro were obtained. Applied conditions for the isolation of luminescent systems (sonication, centrifugation at 40000g) indicate the presence of membrane structures in the extracts, in particular, microsomes formed as a result of ultrasonic disintegration of the endoplasmic reticulum (ER). Differential spectral analysis of the extracts revealed the presence of two absorption peaks at 410 nm and 450 nm, which indicates the presence of cytochromes b5 and P450. The luminescence of the extracts is stimulated by reduced pyridine nucleotides, however, the addition of NADPH causes a higher level of luminescence compared with NADH. The addition of hydrogen peroxide significantly (from several times to 1-2 orders of magnitude) increases the luminescence intensity of extracts activated by NAD(P)H. The addition of fluconazole significantly inhibits the light emission of extracts. The data obtained indicates that the cytochrome P450 system associated with ER membranes may participate in the mechanism of light emission of higher fungi with the involvement in the process of electron transport enzyme systems: NADPH-dependent reductase of cytochrome P450 - cytochrome P450 and NADH-dependent reductase of cytochrome b5 - cytochrome b5 - cytochrome P450. In this case, cytochrome P450 may hydroxylate hispidin (precursor of the luminescent reaction substrate) to form luciferin and catalyze its oxidation in the presence of ROS with light emission.
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