Chun JP, Huber DJ
Horticultural Sciences Department, Institute for Food and Agricultural Sciences, P.O. Box 110690, University of Florida, Gainesville, Florida 32611, USA.
Plant Physiol. 1998 Aug;117(4):1293-9. doi: 10.1104/pp.117.4.1293.
The hydrolysis of cell wall pectins by tomato (Lycopersicon esculentum) polygalacturonase (PG) in vitro is more extensive than the degradation affecting these polymers during ripening. We examined the hydrolysis of polygalacturonic acid and cell walls by PG isozyme 2 (PG2) under conditions widely adopted in the literature (pH 4.5 and containing Na+) and under conditions approximating the apoplastic environment of tomato fruit (pH 6.0 and K+ as the predominate cation). The pH optima for PG2 in the presence of K+ were 1.5 and 0.5 units higher for the hydrolysis of polygalacturonic acid and cell walls, respectively, compared with activity in the presence of Na+. Increasing K+ concentration stimulated pectin solubilization at pH 4.5 but had little influence at pH 6.0. Pectin depolymerization by PG2 was extensive at pH values from 4.0 to 5.0 and was further enhanced at high K+ levels. Oligomers were abundant products in in vitro reactions at pH 4.0 to 5.0, decreased sharply at pH 5.5, and were negligible at pH 6.0. EDTA stimulated PG-mediated pectin solubilization at pH 6.0 but did not promote oligomer production. Ca2+ suppressed PG-mediated pectin release at pH 4.5 yet had minimal influence on the proportional recovery of oligomers. Extensive pectin breakdown in processed tomato might be explained in part by cation- and low-pH-induced stimulation of PG and other wall-associated enzymes.
番茄(Lycopersicon esculentum)多聚半乳糖醛酸酶(PG)在体外对细胞壁果胶的水解作用比果实成熟过程中这些聚合物所受的降解作用更为广泛。我们研究了PG同工酶2(PG2)在文献中广泛采用的条件(pH 4.5且含有Na+)以及接近番茄果实质外体环境的条件(pH 6.0且以K+作为主要阳离子)下对聚半乳糖醛酸和细胞壁的水解情况。与在Na+存在时的活性相比,在K+存在时PG2对聚半乳糖醛酸和细胞壁水解的最适pH分别高出1.5和0.5个单位。增加K+浓度在pH 4.5时刺激果胶溶解,但在pH 6.0时影响很小。PG2在pH值4.0至5.0时对果胶的解聚作用广泛,在高K+水平下进一步增强。在pH 4.0至5.0的体外反应中,寡聚物是丰富的产物,在pH 5.5时急剧减少,在pH 6.0时可忽略不计。EDTA在pH 6.0时刺激PG介导的果胶溶解,但不促进寡聚物的产生。Ca2+在pH 4.5时抑制PG介导的果胶释放,但对寡聚物的比例回收率影响最小。加工番茄中广泛的果胶分解可能部分归因于阳离子和低pH对PG及其他细胞壁相关酶的刺激作用。