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通过β-胡萝卜素的酶促中心裂解在体外无法形成视黄醛。

Retinal is not formed in vitro by enzymatic central cleavage of beta-carotene.

作者信息

Hansen S, Maret W

机构信息

Fachbereich 15.2, Analytische und Biologische Chemie, Universität des Saarlandes, Saarbrücken, FRG.

出版信息

Biochemistry. 1988 Jan 12;27(1):200-6. doi: 10.1021/bi00401a030.

Abstract

Rat intestinal mucosa was prepared and incubated with beta-carotene by the procedure of Goodman and Olson [Goodman, DeW. S., & Olson, J.A. (1969) Methods Enzymol. 15, 462-475] to determine beta-carotene cleavage activity. A new detection system for the reaction products of the described enzyme beta-carotene 15,15'-dioxygenase (EC 1.13.11.21) employs solvent extraction of retinoids and carotenoids followed by high-performance liquid chromatography separation and photometric detection of the pigments. It has not detected any newly formed retinal or other retinoids in the intestinal protein preparations from normal or vitamin A deficient rats. The latter were chosen as a possible source of more active enzyme preparations. With corresponding blank samples subjected to identical conditions of incubation but without added protein, small amounts of beta-apocarotenals could be detected. They were previously reported as cleavage products of beta-carotene [Ganguly, J., & Sastry, P.S. (1985) World Rev. Nutr. Diet. 45, 198-220] but are clearly not formed as a result of an enzymatic reaction. The failure to detect in vitro enzymatic central or random cleavage of the beta-carotene molecule in extracts of rat intestinal mucosa emphasizes the need to reevaluate the existing theory of conversion of beta-carotene into vitamin A.

摘要

按照古德曼和奥尔森的方法[古德曼,德W. S.,&奥尔森,J. A.(1969年)《酶学方法》15,462 - 475]制备大鼠肠黏膜并与β - 胡萝卜素一起孵育,以测定β - 胡萝卜素裂解活性。一种用于所述β - 胡萝卜素15,15'-双加氧酶(EC 1.13.11.21)反应产物的新检测系统,采用类视黄醇和类胡萝卜素的溶剂萃取,随后进行高效液相色谱分离以及色素的光度检测。在正常或维生素A缺乏大鼠的肠蛋白制剂中,未检测到任何新形成的视黄醛或其他类视黄醇。选择后者作为可能产生更具活性酶制剂的来源。在相同孵育条件下但不添加蛋白质的相应空白样品中,可检测到少量的β - 阿朴胡萝卜醛。它们先前被报道为β - 胡萝卜素的裂解产物[甘古利,J.,&萨斯特里,P. S.(1985年)《世界营养与饮食评论》45,198 - 220],但显然不是酶促反应的结果。在大鼠肠黏膜提取物中未能检测到β - 胡萝卜素分子的体外酶促中心或随机裂解,这突出表明需要重新评估现有的β - 胡萝卜素转化为维生素A的理论。

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