Hamamoto T, Lee Y C, Kurosawa N, Nakaoka T, Kojima N, Tsuji S
Glyco Molecular Biology, Frontier Research Program, Institute of Physical and Chemical Research (RIKEN), Saitama, Japan.
Bioorg Med Chem. 1994 Feb;2(2):79-84. doi: 10.1016/s0968-0896(00)82004-0.
Mouse Gal beta 1,4GlcNAc alpha 2,6-sialyltransferase was produced in an insoluble form in Escherichia coli cells harboring expression plasmids. The insoluble protein was solubilized with 8 M urea and diluted for renaturation of the enzyme. The substrate specificity and kinetic parameters, except for the specific activity, of the renatured enzyme were similar to those of the enzyme obtained from rat liver. These results suggest that a bacterial expression system is a potentially powerful tool for the large scale production of sialyltransferases and for elucidating the molecular mechanisms of sialyltransferases.
小鼠Galβ1,4GlcNAcα2,6-唾液酸转移酶在携带表达质粒的大肠杆菌细胞中以不溶性形式产生。用8M尿素溶解不溶性蛋白，并稀释以进行酶的复性。复性酶的底物特异性和动力学参数（除比活性外）与从大鼠肝脏获得的酶相似。这些结果表明，细菌表达系统是大规模生产唾液酸转移酶和阐明唾液酸转移酶分子机制的潜在有力工具。
