Metabolism of d-glucuronolactone in mammalian systems. Identification of d-glucaric acid as a normal constituent of urine.
Metabolism of d-glucuronolactone in mammalian systems. Identification of d-glucaric acid as a normal constituent of urine.
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哺乳动物系统中 d-葡萄糖醛酸内酯的代谢。
DOI：
10.1042/bj0860077
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发表时间：
1963
期刊：
影响因子：
--
通讯作者：
C. Marsh
中科院分区：
文献类型：
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作者：
C. Marsh
Aberdeen皇家医院的mamResearch Unit对P-葡萄糖醛酸酶的抑制作用，许多作者注意到了马里尿。新鲜的猫尿样本是工人的。这使A博士提供的酶的测定复杂化。J.卡尔，病理学系，Aberurine，和添加的 *-e的使用;大鼠和豚鼠的24小时尿液，以及绵羊、猪和小牛尿液的尿液和β-葡萄糖醛酸酶1的使用，从在代谢笼中维持正常饮食的动物中收集。从β-D-葡糖苷醛酸中分离出甾体化合物。β-葡萄糖醛酸酶测定法通常是将粪便用水洗涤，然后与显色底物结合，对酶尿和洗涤液进行检测。如果尿液的水解在采集后24小时内可能会减少，则将尿液储存在0 °，否则储存在20°。存在含有竞争性尿液亚处理的尿液。以下是不同亲和力的标准程序底物（参见Levvy & Marsh，用于测试哺乳动物的抑制能力1954）。除了尿的天然底物外。用1000葡萄糖醛酸酶处理粗尿（pH 4-5-7-0），然而，用3 N-HCl调节pH 2-0-2-2后，将尿液浓缩40分钟，然后用NaOH调节pH 4-0-45，或将其转化为真正的酶抑制剂。Abul-Fadl（1957）（B）在100°下15分钟，用0-1 N NaOH调节后发现可透析和不可透析物质的pH值均为7-5-8-0，随后用HCl重新调节至人尿液中的pH值60，这对β-gluc 6 -5具有抑制作用。糖醛酸酶处理后抑制力最大;可透析抑制剂对（a）稳定，处理后最小（B）。两种处理均为热酸处理。兔尿中也存在类似的抑制组分，足以消除尿液中的β-葡萄糖醛酸酶活性（Conzelman &重新调整后，对Crout无干扰，1961）。后续酶测定的pH。在哺乳动物β-葡萄糖醛酸酶被抑制2-5以上的pH下加热不足以产生最大的重金属离子（Levvy & Marsh，1957 a，B;酸化尿液的抑制能力，因为那时有一个
The inhibition of P-glucuronidase by mamResearch Unit, Aberdeen Royal Infirmary, to whom the malian urine has been noted by a number of author is indebted. Fresh samples of cat urine were workers. This complicates assay of the enzyme in supplied by Dr A. J. Carr, Pathology Department, Aberurine, and the use of added *-e to deen University; 24 hr. urines of rat and guinea pig, and urine, and the us of -glucuronidase 1 samples of sheep, pig and calf urines, were collected from lhberate excretedl phenols and alcohols' includm' let xo s, i ng animals maintained on normal diets in metabolism cages. steroids, from the fl-D-glucosiduronic acid conThe rat and guinea-pig exereta were separated by filtration, jugates. P-Glucuronidase assay is normally conand the faeces washed with water, before the combined ducted with a chromogenic substrate, the enzymic urine and washings were tested. Urines were stored at 0' if hydrolysis of which may be diminished in the examined within 24 hr. of collection, or otherwise at 20°. presence of urine which contains competing subTreatment of urine. The following standard procedure strates of different affinities (see Levvy & Marsh, was used in testing the inhibitory powers of mammalian 1954). In addition to natural substrates for p urine. Crude urine (pH 4-5-7-0) was treated (a) at 1000 glucuronidase, however, mammnalian urine confor 40 min. after adjustment with 3N-HCl to pH 2-0-2-2, tains trueenzymeinhibitrs Abul-Fadl(1957) followed by readjustment with NaOH to pH 4-0-45, or tamns true enzyme inhibitors. Abul-Fadl (1957) (b) at 100° for 15 min. after adjustment with 0-1 N-NaOH to found both dialysable and non-dialysable material pH 7-5-8-0, follQwed by readjustment with HCI to pH 60in human urine which was inhibitory to /-gluc6-5. The inhibitory power was maximal after treatment uronidase; the dialysable inhibitor was stable to (a) and minimal after treatment (b). Either treatment was treatment with hot acid. Similar inhibitory fracsufficient to rid the urine of ,B-glucuronidase activity, and tions were present in rabbit urine (Conzelman & after the readjustments there was no interference with the Crout, 1961). pH of the subsequent enzyme assay. Heating at a pH Mammalian /3-glucuronidase is inhibited by above 2-5 was inadequate for development of maximum heavy-metal ions (Levvy & Marsh, 1957a, b; inhibitory power of acidified urines, for there was then a