Partial purification and properties of alanine racemase from the muscle of black tiger prawn Penaeus monodon.
Partial purification and properties of alanine racemase from the muscle of black tiger prawn Penaeus monodon.
复制标题
斑节对虾肌肉丙氨酸消旋酶的部分纯化及其性质。
DOI：
10.2331/fishsci.63.440
复制
发表时间：
1997
影响因子：
1.9
通讯作者：
H. Abe
中科院分区：
文献类型：
--
作者：
E. Fujita;E. Okuma;H. Abe
采用DEAE-纤维素、DEAE Toyopearl、羟基磷灰石、Phenyl和Butyl-Toyopearl以及Gel-Toyopearl HW柱层析从黑虎虾斑节对虾肌肉中纯化。最终的酶制剂不是均相的，但纯化高达约17，700倍，最终产率为2.5%。天然形式的酶的表观分子量为85，000。在35-40 ℃和pH 8.5左右达到最大活性。丙氨酸消旋酶在40 ~ 60 ℃范围内失活，在低温贮藏时也不稳定。该酶特异性地作用于作为底物的D-、L丙氨酸，但在本测定条件下不作用于其它氨基酸。该酶不需要吡哆醛5·β-磷酸或FAD作为辅因子。D-丙氨酸的代谢产物丙酮酸和L-丙氨酸强烈抑制该酶。
purified from the muscle of the black tiger prawn Penaeus monodon using DEAE-cellulose, DEAE Toyopearl, hydroxyapatite, Phenyl and Butyl-Toyopearl, and Gel-Toyopearl HW column chro matographies. The final enzyme preparation was not homogeneous but the purification was as high as about 17,700-fold with a final yield of 2.5%. Apparent molecular weight of the enzyme in its native form was 85,000. The maximal activity was attained at 35-40•Ž and at around pH 8.5. The alanine racemase was inactivated between 40 and 60•Ž and was also rather unstable during low temperature storage. The enzyme acts specifically on D-, L alanine as substrates, but not on the other amino acids in the present assay conditions. The enzyme did not require pyridoxal 5•Œ-phosphate or FAD as a cofactor. The enzyme was inhibited strongly with pyru vate and L-alanine, which are metabolites from D-alanine.