Appl Biochem Biotechnol. 2013 Dec;171(8):2040-52. doi: 10.1007/s12010-013-0491-9.
The purification and characterization of intracellular tyrosine phenol lyase from Citrobacter freundii has been carried out. The enzyme was purified 35-fold to homogeneity by ammonium sulphate precipitation and hydrophobic interaction chromatography. Its subunit molecular weight was found to be 52 kDa on sodium dodecyl sulphate polyacrylamide gel electrophoresis. The purified tyrosine phenol lyase showed maximum activity in borate buffer (0.05 M at pH 8.5) at 45 °C after 20 min of incubation. The Km and Vmax values of purified enzyme were found to be 0.446 mm and 0.342 mM/min/mg. This enzyme exhibits t1/2 of 10, 52 and 130 min at 55, 45 and 35 °C, respectively. The N-terminal amino acid sequence was determined as MET-ASN-TYR-PRO-ALA-GLU-PRO-PHE-ARG-ILETRP- TRP-VAL-GLY.
已对弗氏柠檬酸杆菌细胞内酪氨酸酚裂解酶进行了纯化和特性分析。该酶通过硫酸铵沉淀和疏水相互作用层析进行了 35 倍的纯化，达到了均一性。在十二烷基硫酸钠聚丙烯酰胺凝胶电泳中，其亚基分子量为 52 kDa。纯化后的酪氨酸酚裂解酶在硼酸缓冲液（pH 8.5 的 0.05 M）中，在 45°C 下孵育 20 分钟后表现出最大活性。该酶的 Km 和 Vmax 值分别为 0.446 mm 和 0.342 mM/min/mg。该酶在 55°C、45°C 和 35°C 下的 t1/2 值分别为 10、52 和 130 分钟。N 端氨基酸序列测定为 MET-ASN-TYR-PRO-ALA-GLU-PRO-PHE-ARG-ILETRP-TRP-VAL-GLY。
