Appl Environ Microbiol. 1995 Jan;61(1):21-6. doi: 10.1128/aem.61.1.21-26.1995.
Dimethyl sulfide (DMS) is quantitatively the most important biogenic sulfur compound emitted from oceans and salt marshes. It is formed primarily by the action of dimethylsulfoniopropionate (DMSP) lyase which cleaves DMSP, an algal osmolyte, to equimolar amounts of DMS and acrylate. This report is the first to describe the isolation and purification of DMSP lyase. The soluble enzyme was purified to electrophoretic homogeneity from a facultatively anaerobic gram-negative rod-shaped marine bacterium identified as an Alcaligenes species by the Vitek gram-negative identification method. The key to successful purification of the enzyme was its binding to, and hydrophobic chromatography on, a phenyl-Sepharose CL-4B column. DMSP lyase biosynthesis was induced by its substrate, DMSP; its product, acrylate; and also by acrylamide. The relative effectivenesses of the inducers were 100, 90, and 204%, respectively. DMSP lyase is a 48-kDa monomer with a Michaelis-Menten constant (K(infm)) for DMSP of 1.4 mM and a V(infmax) of 408 (mu)mol/min/mg of protein. It converted DMSP to DMS and acrylate stoichiometrically. The similar K(infm) values measured for pure DMSP lyase and the axenic culture, seawater, and surface marsh sediment suggest that the microbes in these ecosystems must have enzymes similar to the one purified from our marine isolate. Anoxic sediment populations, however, have a 40-fold-lower K(infm) for this enzyme (30 (mu)M), possibly giving them the capability to metabolize much lower levels of DMSP than the aerobes.
二甲基硫(DMS)是海洋和盐沼中排放量最大的生物源硫化合物,其生成主要是通过二甲基巯基丙酸酯(DMSP)裂解酶的作用,该酶能将 DMSP——一种藻类渗透调节剂——切割成等摩尔量的 DMS 和丙烯酸盐。本报告首次描述了 DMSP 裂解酶的分离和纯化。该可溶性酶是从一种兼性厌氧革兰氏阴性棒状海洋细菌中分离得到的,通过 Vitek 革兰氏阴性鉴定方法鉴定为产碱杆菌属。成功纯化该酶的关键是其与苯基-Sepharose CL-4B 柱的结合及其疏水性色谱。DMSP 裂解酶的生物合成受其底物 DMSP、产物丙烯酸盐和丙烯酰胺的诱导。这些诱导剂的相对有效性分别为 100%、90%和 204%。DMSP 裂解酶是一种 48kDa 的单体,其对 DMSP 的米氏常数(K(infm))为 1.4mM,最大反应速度(V(infmax))为 408(mu)mol/min/mg 蛋白。它将 DMSP 转化为 DMS 和丙烯酸盐,具有化学计量关系。纯 DMSP 裂解酶和无菌培养物、海水和表层沼泽沉积物的 K(infm)值相似,这表明这些生态系统中的微生物必须具有与我们从海洋分离株中纯化的酶相似的酶。然而,缺氧沉积物中的这种酶的 K(infm)值低 40 倍(30(mu)M),这可能使它们能够代谢比好氧微生物低得多水平的 DMSP。