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Is-beta-alanine-specific opine dehydrogenase present?

是否存在β-丙氨酸特异性阿片脱氢酶？

基本信息

批准号：
08660258
负责人：
KAN-NO Nobuhiro
金额：
$ 1.54万
依托单位：
Kitasato University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

项目摘要

Distribution of opin production pathway was examined for a wide variety of marine invertebrates by a combination of activities of opine dehydrogenases (OpDH), band patterns on isoelectric focusing in polyacrylamide gel electrophoresis (IEF-PAG), and levels of free amino acids. In several animals, significant beta-alanopine dehydrogenase (beta-AIDH) activity was observed. However, there was no case that beta-AIDH activity alone was found in the species. beta-AIDH activity was found as a dominant OpDH activity in the two species of limpets. However, in many other species, b-ALDH activity was found rather minor OpDH activity. In those animals, higher activities of alanopine dehydrogenase (AIDH) and/or strombine dehydrogenase (StDH) in addition to beta-AIDH activity was observed. IEF-PAG analysis showed that band patterns for beta-AIDH activity and AIDH activity were nearly identical, suggesting that later enzyme was a broadly specific OpDH showing beta-AIDH activity. OpDHs were purified a … More nd characterized from the limpet Cellana grata and the gastropod Fusitriton oregonensis. The muscular tissues of the lipet C.geata exhibited beta-AIDH activity in addition to TaDH activity and weak lactate dehydrogenase activity. OpDHs were purified, and two different types of OpDH,TaDH and OpDH showing beta-AIDH activity, were isolated. From the specificity for amino acid and opine, OpDH showing beta-AIDH activity was concluded to be a true beta-AIDH showing strict substrate specificity for beta-alanine. While the catalytic properties of the two enzymes were essentially similar, they were distinct from each other with respect to the amino acid substrate specificity and the K_m values. Great similarities were found between beta-AIDH and TaDH with respect to molecular properties. These results clearly showed that beta-alanine specific OpDH,true beta-AIDH,was present in the limpet muscle. On the other hand, activities of AIDH,StDH,beta-AIDH,and TaDH were observed in the muscle of F.oregonensis. Four enzymes were isolated. The characterization study showed that F.oregonensis OpDHs were broadlyspecific enzymes. The catalytic properties of OpDHs showed that beta-AIDH activity had no physiological mean. Thus it was concluded that true beta-AIDH was present, but it was also true that all of beta-AIDH activity detected in invertebrate did not indicated the presence of beta-alanopine formation pathway in the animal. Less

通过结合OpDH活性、聚丙烯酰胺凝胶电泳（IEF-PAG）等电聚焦带模式和游离氨基酸水平，研究了多种海洋无脊椎动物的opin生产途径分布。在一些动物中，观察到显著的β -丙氨酸脱氢酶（β - aidh）活性。然而，在该物种中没有发现单独的β - aidh活性。在两种帽贝中发现- aidh活性是显性的OpDH活性。然而，在许多其他物种中，发现b-ALDH活性相对较小的OpDH活性。在这些动物中，除了β -AIDH活性外，还观察到较高的丙氨酸脱氢酶（AIDH）和/或基质脱氢酶（StDH）活性。IEF-PAG分析显示，β -AIDH活性和AIDH活性的条带模式几乎相同，表明后一种酶是具有β -AIDH活性的广泛特异性OpDH。从帽贝和俄勒冈赤足动物中纯化和鉴定了OpDHs。除TaDH活性外，脂肪肌肉组织还表现出β - aidh活性和较弱的乳酸脱氢酶活性。对OpDH进行纯化，分离得到两种不同类型的OpDH，TaDH和具有β - aidh活性的OpDH。从对氨基酸和视氨酸的特异性来看，显示β - aidh活性的OpDH是对β -丙氨酸具有严格底物特异性的真正的β - aidh。虽然两种酶的催化性能基本相似，但它们在氨基酸底物特异性和K_m值方面存在差异。β - aidh和TaDH在分子性质上有很大的相似之处。这些结果清楚地表明，在帽贝肌中存在β -丙氨酸特异性OpDH，即真正的β - aidh。另一方面，在赤羊肌肉中观察到AIDH、StDH、β -AIDH和TaDH的活性。分离到4种酶。表征研究表明，oregonensis OpDHs具有广泛的特异性。OpDHs的催化性能表明β - aidh活性没有生理平均值。因此，我们得出结论，确实存在β - aidh，但在无脊椎动物中检测到的所有β - aidh活性并不表明动物中存在β -丙氨酸形成途径。少