Kobayashi G, Toida J, Akamatsu T, Yamamoto H, Shida T, Sekiguchi J
Department of Applied Biology, Faculty of Textile Science and Technology, Shinshu University, 3-15-1 Tokida, Ueda, Nagano 386-8567, Japan.
J Biosci Bioeng. 2000;90(4):422-5.
cutL cDNA encoding an extracellular lipase, L1, from Aspergillus oryzae was fused to the cell wall-binding domain (CWB) region of a plasmid, pHCB3R. SDS-polyacrylamide gel electrophoresis (PAGE) and zymography of proteins extracted from the cell surface of Bacillus subtilis 168 harboring a fused lipase plasmid (pHCB3RCL) revealed that the fused gene product, CWB-CutL, was localized in the B. subtilis cell wall and retained lipase activity. B. subtilis WASD (wprA sigD), recently used for the accumulation of CWB-LipB (the CWB protein fused with B. subtilis lipase B), was also a suitable host for the accumulation of CWB-CutL, the amount being 10% of the total proteins extracted from the cell surface.
编码来自米曲霉的一种细胞外脂肪酶L1的cutL cDNA与质粒pHCB3R的细胞壁结合结构域（CWB）区域融合。对携带融合脂肪酶质粒（pHCB3RCL）的枯草芽孢杆菌168细胞表面提取的蛋白质进行SDS-聚丙烯酰胺凝胶电泳（PAGE）和酶谱分析，结果显示融合基因产物CWB-CutL定位于枯草芽孢杆菌细胞壁并保留脂肪酶活性。最近用于积累CWB-LipB（与枯草芽孢杆菌脂肪酶B融合的CWB蛋白）的枯草芽孢杆菌WASD（wprA sigD），也是积累CWB-CutL的合适宿主，其含量占从细胞表面提取的总蛋白的10%。
