PURIFICATION AND PROPERTIES OF STRICTOSIDINE SYNTHASE, THE KEY ENZYME IN INDOLE ALKALOID FORMATION
PURIFICATION AND PROPERTIES OF STRICTOSIDINE SYNTHASE, THE KEY ENZYME IN INDOLE ALKALOID FORMATION
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DOI：
10.1111/j.1432-1033.1979.tb04235.x
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发表时间：
1979-01-01
期刊：
影响因子：
--
通讯作者：
ZENK, MH
中科院分区：
文献类型：
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作者：
TREIMER, JF;ZENK, MH
从长春花悬浮培养细胞的可溶性蛋白提取物中分离得到一种新的催化色胺和番泻素合成3-α-S的新酶，该酶经硫酸铵分级沉淀、DEAE-纤维素柱层析、超凝胶AcA34和等电聚焦等步骤纯化约50倍。该酶的表观分子量为34,000。该酶合成斯特拉西丁的最适pH为6.8，色胺和防己内酯的表观Km值分别为2.3 mM和3.4 mM。马钱子苷合成酶具有很高的底物专一性。没有明显的辅因要求被证明。在测试的几种酶抑制剂中，只有对氯汞苯甲酸盐抑制该酶。该酶相对稳定，可在-20℃下保存。C，最长可达一年，没有明显的催化活性损失。该酶存在于夹竹桃科产吲哚生物碱亚科Plumerioideae的9个不同属的15个种的悬浮培养中[Amsia salicifolia、A.taberna ememontana、长叶长春花、C.pusillus、C.roseus、C.trichop llus、Ervatamia divaricata、Ochrosia Eliptica、Rauwolfia verticillata、R.vomitoria、Rhazya orientalis、Stemmdenia tomentosa Palmeri、Vinca main、V.Minor和Voacanga africana]。这种酶负责合成严格的苦苷，这是植物界中大多数单萜类吲哚生物碱形成的关键中间体。
A new enzyme, strictosidine synthase, which catalyzes the synthesis of 3-.alpha.(S)-strictosidine from tryptamine and secologanin was isolated from the soluble protein extract of Catharanthus roseus [= Vinca rosea] cell suspension cultures and was purified approximately 50-fold by ammonium sulfate fractionation, column chromatography on DEAE-cellulose, Ultrogel AcA34 and isoelectric focusing. The apparent MW of the enzyme was 34,000. The pH optimum was 6.8, apparent Km values for tryptamine and secologanin were 2.3 mM and 3.4 mM, respectively, for the enzyme to synthesize strictosidine. Strictosidine synthase shows high substrate specificity. No apparent cofactor requirement could be demonstrated. Of several enzyme inhibitors tested, only p-chloromercuribenzoate inhibited the enzyme. The enzyme was relatively stable and could be stored at -20.degree. C for periods of up to 1 yr without appreciable loss of catalytic activity. The enzyme was demonstrated to occur in suspension cultures of 15 different species [Amsonia salicifolia, A. tabernaemontana, Catharanthus longifolius, C. pusillus, C. roseus, C. trichophyllus, Ervatamia divaricatum, Ochrosia eliptica, Rauwolfia verticillata, R. vomitoria, Rhazya orientalis, Stemmadenia tomentosa palmeri, Vinca major, V. minor and Voacanga africana] belonging to 9 different genera of the indole-alkaloid-producing subfamily Plumerioideae of the Apocynaceae family. This enzyme is responsible for the synthesis of strictosidine the key intermediate in the formation of the majority of monoterpenoid indole alkaloids occurring in the plant kingdom.