Sakamoto T, Hours R A, Sakai T
Department of Agricultural Chemistry, College of Agriculture, University of Osaka Prefecture, Japan.
Biosci Biotechnol Biochem. 1994 Feb;58(2):353-8. doi: 10.1271/bbb.58.353.
We found two enzymes that solubilize pectin from protopectin, tentatively named protopectinase-N (PPase-N) and protopectinase-R (PPase-R), in a culture filtrate of Bacillus subtilis IFO 3134. These enzymes were purified to homogeneity by hydrophobic, cation exchange and size exclusion chromatographies. The molecular weights of PPase-N and PPase-R were estimated to be 43,000 and 35,000, respectively, by SDS-PAGE. Their pIs were 9.4 and 8.2, respectively. These enzymes were stable in a wide range of pH and temperature. PPase-N and -R released water-soluble pectin by transeliminative cleavage of protopectin. According to their substrate specificities and modes of action, PPase-N and PPase-R could be classified as endo-pectate transeliminase (pectate lyase; EC 4.2.2.2) and endo-pectin transeliminase (pectin lyase; EC 4.2.2.10), respectively. Both enzymes were produced in a simple medium containing defatted soybean flour and phosphates. Production of PPase-N was repressed by addition of glucose while that of PPase-R was enhanced by phosphate.
我们在枯草芽孢杆菌IFO 3134的培养滤液中发现了两种可将原果胶中的果胶溶解的酶，暂命名为原果胶酶-N（PPase-N）和原果胶酶-R（PPase-R）。通过疏水色谱、阳离子交换色谱和尺寸排阻色谱将这些酶纯化至均一。通过SDS-PAGE估计PPase-N和PPase-R的分子量分别为43,000和35,000。它们的pI分别为9.4和8.2。这些酶在较宽的pH和温度范围内都很稳定。PPase-N和-R通过原果胶的反式消除裂解释放水溶性果胶。根据它们的底物特异性和作用方式，PPase-N和PPase-R可分别归类为内切聚半乳糖醛酸反式消除酶（聚半乳糖醛酸裂解酶；EC 4.2.2.2）和内切果胶反式消除酶（果胶裂解酶；EC 4.2.2.10）。这两种酶都在含有脱脂大豆粉和磷酸盐的简单培养基中产生。添加葡萄糖会抑制PPase-N的产生，而磷酸盐会增强PPase-R的产生。
