Separation of Proteinase from Embryos during Incubation of chicken Eggs and Control of the Activity by Endogenous Proteinase Inhibitor

两种蛋白质酶的数量很小（A和B）从50只鸡蛋的胚胎中分离出来，在第七天的孵化中，通过对ovoinhibitor-Sepharose和ovomucoid-Sepharose柱的反应性亲和色谱法。这些酶的活动通常非常弱，也可能几乎可以用Kunitz的外壳消化方法来确定反应时间为4小时或5小时。酶A的最佳pH值从8到10不等，酶B的值接近6.0。这些酶的最佳温度是42 ° C。Enzyme一种活动不受Mg、Cuion和EDTA的添加的影响，但受Fe^<2+>或Zn离子（10^<-3>M）的影响。45%;酶是由二异丙基氟磷酸盐修饰而不激活的，由内源性ovoinhibitor抑制，未由ovomucoid抑制。酶，变形，不是氢化的p-tosyl arg。甲基化，但氢化的苯唑醇。ethylester。从这些结果中，酶A被假定为类似酶的酶。从酶A和B中不同，酶A和B没有被ovoinhibitor抑制，但被ovomucoid抑制;它不是氢化BTEE，而是氢化TAME。因此，酶B被认为是trypsin--就像酶一样。这些酶在化学物质中有所不同。两种酶都是氢化的，非常缓慢地分解的蛋白解决方案（conc.）4%）加热后，内源性抑制剂（ovo-inhibitor和ovomucoid）的抑制作用。这些实验性结果提出了由内基因蛋白酶抑制剂在鸡胚胎生长期间对蛋白质酸的控制或调节。

Very small amounts of two kinds of proteinases (A and B) were separated from the extract of embryos of fifty chicken eggs at the 7th day of incubation by the respective affinity chromatography on ovoinhibitor-Sepharose and ovomucoid-Sepharose columns. The activities of these enzymes were respectively very weak and could be barely assayed using kunitz's casein digestion method by taking the reacting time of 4 or 5 hr. Optimum pH value of enzyme A for the reation was ranging from 8 to 10 and thet of enzyme B was approximately 6.0. Optimum temeratures of these enzymes were respectively 42゜C. Enzyme A activity was not affected by addition of Mg, Cu ions and EDTA, but inhibited by that of Fe^<2+> or Zn ion (10^<-3>M) by ca. 45 %. The enzyme was inactivated by modification with diisopropylflurophosphate and inhibited by endogenous ovoinhibitor, and not inhibited by ovomucoid. The enzyme, moreover, did not hydrolyzed p-tosyl arg.-methylester, but hydrolyzed benzoyltyr.-ethylester. From these results, enzyme A was presumed to be chymotrypsin-like enzyme.Differing from enzyma A, B was not inhibited by ovoinhibiter, but inhibited by ovomucoid; it did not hydrolyze BTEE, but hydrolyzed TAME. Therefore, enzyme B was presumed to be trypsin-like enzyme. These enzymes differed in the chemical constitutions. Both the enzymes hydrolyed very slowly diluted egg white solutions (concn.: 4 %) after inactivation of endogenous inhibitors (ovo-inhibitor and ovomucoid) by heating. These experimental results suggest the control or regulation of proteolysis by endogenous proteinase inhibitors during chicken embryonic growth.