Dmitryjuk M, Dopieralska M, Łopieńska-Biernat E, Frączek R J
Department of Biochemistry, Faculty of Biology, University of Warmia and Mazury, Olsztyn, Poland.
J Helminthol. 2013 Jun;87(2):212-21. doi: 10.1017/S0022149X12000259. Epub 2012 May 9.
Trehalose 6-phosphate (T6P) synthase (TPS; EC 2.4.1.15) was isolated from muscles of Ascaris suum by ammonium sulphate fractionation, ion-exchange DEAE SEPHACEL(TM) anion exchanger column chromatography and Sepharose 6B gel filtration. On sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), 265-fold purified TPS exhibited a molecular weight of 66 kDa. The optimum pH and temperature of the purified enzyme were 3.8-4.2 and 35°C, respectively. The isoelectric point (pI) of TPS was pH 5.4. The studied TPS was not absolutely substrate specific. Besides glucose 6-phosphate, the enzyme was able to use fructose 6-phosphate as an acceptor of glucose. TPS was activated by 10 mM MgCl2, 10 mM CaCl2 and 10 mM NaCl. In addition, it was inhibited by ethylenediaminetetra-acetic acid (EDTA), KCl, FeCl3 and ZnCl2. Two genes encoding TPS were isolated and sequenced from muscles of the parasite. Complete coding sequences for tps1 (JF412033.2) and tps2 (JF412034.2) were 3917 bp and 3976 bp, respectively. Translation products (AEX60788.1 and AEX60787.1) showed expression to the glucosyltransferase-GTB-type superfamily.
通过硫酸铵分级分离、离子交换DEAE SEPHACEL™阴离子交换柱色谱法和琼脂糖6B凝胶过滤，从猪蛔虫肌肉中分离出海藻糖6-磷酸（T6P）合酶（TPS；EC 2.4.1.15）。在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）上，经过265倍纯化的TPS呈现出66 kDa的分子量。纯化酶的最适pH和温度分别为3.8 - 4.2和35°C。TPS的等电点（pI）为pH 5.4。所研究的TPS并非绝对底物特异性。除了6-磷酸葡萄糖外，该酶还能够使用6-磷酸果糖作为葡萄糖的受体。TPS被10 mM MgCl2、10 mM CaCl2和10 mM NaCl激活。此外，它受到乙二胺四乙酸（EDTA）、KCl、FeCl3和ZnCl2的抑制。从该寄生虫的肌肉中分离并测序了两个编码TPS的基因。tps1（JF412033.2）和tps2（JF412034.2）的完整编码序列分别为3917 bp和3976 bp。翻译产物（AEX60788.1和AEX60787.1）显示属于糖基转移酶-GTB型超家族。
