细菌L-甲硫氨酸γ-裂解酶的纯化
Purification of bacterial L-methionine gamma-lyase.
作者信息
Nakayama T, Esaki N, Sugie K, Beresov T T, Tanaka H, Soda K
出版信息
Anal Biochem. 1984 May 1;138(2):421-4. doi: 10.1016/0003-2697(84)90832-7.
A rapid procedure for the purification of L-methionine gamma-lyase from Pseudomonas putida ICR 3460 by DEAE- TOYOPEARL 650M and DEAE-Sephadex A-50 column chromatography is presented. The enzyme was purified with an average yield of 75% and showed about 10-fold higher specific activity than the enzyme from P. putida (= P. ovalis) IFO 3738 reported previously (H. Tanaka, N. Esaki , and K. Soda (1976) FEBS Lett. 66, 307-311). The present enzyme has a molecular weight of about 172,000 and consists of four subunits with identical molecular weights (43,000). It shows the typical absorption spectrum of pyridoxal enzyme with maxima at 278 and 420 nm, and contains 4 mol of pyridoxal 5'-phosphate per mole of enzyme. The enzyme has a multicatalytic function similar to the enzyme of P. putida IFO 3738 (K. Soda, H. Tanaka, and N. Esaki (1983) Trends Biochem. Sci. 8, 214-217).
本文介绍了一种通过DEAE - TOYOPEARL 650M和DEAE - Sephadex A - 50柱色谱法从恶臭假单胞菌ICR 3460中快速纯化L - 蛋氨酸γ - 裂合酶的方法。该酶的纯化平均产率为75%，比先前报道的恶臭假单胞菌（=卵形假单胞菌）IFO 3738的酶的比活性高约10倍（H. Tanaka、N. Esaki和K. Soda（1976年）《欧洲生物化学学会联合会快报》66，307 - 311）。目前的这种酶分子量约为172,000，由四个分子量相同（43,000）的亚基组成。它显示出吡哆醛酶的典型吸收光谱，在278和420 nm处有最大值，每摩尔酶含有4摩尔吡哆醛5'-磷酸。该酶具有与恶臭假单胞菌IFO 3738的酶类似的多催化功能（K. Soda、H. Tanaka和N. Esaki（1983年）《生物化学趋势》8，214 - 217）。
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