McNab J M, Villemez C L, Albersheim P
Biochem J. 1968 Jan;106(2):355-60. doi: 10.1042/bj1060355.
A particulate cell-free enzyme system was prepared from Phaseolus aureus shoots. This preparation was able to incorporate [(14)C]galactose from UDP-[(14)C]galactose into a water-soluble polysaccharide, which has a probable molecular weight of at least 4600. The only labelled component detectable in the polymer was shown to be [(14)C]galactose; two labelled oligosaccharides containing only [(14)C]galactose were isolated by partial hydrolysis. The galactan-synthesizing activity of this particulate preparation is maximal at 30 degrees and pH7.1 in the presence of 5.0mm-magnesium chloride and 0.2m-sucrose. Although 3-day-old seedlings were used as a source of enzyme, it appears that 4- or 5-day-old beans contain greater synthetase activity. The enzyme system has an apparent Michaelis constant of 5.8x10(-6)m, and will catalyse the polymerization of galactose residues at the rate of 7.5mmumoles/mg. of protein/min. at a substrate concentration of 9.6mm.
从金甲豆嫩枝中制备了一种无细胞颗粒酶系统。该制剂能够将UDP - [(14)C]半乳糖中的[(14)C]半乳糖掺入一种水溶性多糖中,该多糖的分子量可能至少为4600。聚合物中唯一可检测到的标记成分显示为[(14)C]半乳糖;通过部分水解分离出两种仅含有[(14)C]半乳糖的标记寡糖。这种颗粒制剂的半乳聚糖合成活性在30摄氏度和pH7.1、存在5.0毫摩尔/升氯化镁和0.2摩尔/升蔗糖的条件下最大。虽然使用3日龄幼苗作为酶源,但似乎4日龄或5日龄的豆子含有更高的合成酶活性。该酶系统的表观米氏常数为5.8×10(-6)摩尔/升,在底物浓度为9.6毫摩尔/升时,将以7.5微摩尔/毫克蛋白质/分钟的速率催化半乳糖残基的聚合。