Sakakibara R, Uemura M, Hirata T, Okamura N, Kato M
Department of Biochemistry, School of Pharmaceutical Sciences, Nagasaki University, Japan.
Biosci Biotechnol Biochem. 1997 Nov;61(11):1949-52. doi: 10.1271/bbb.61.1949.
The nucleotide sequence of 1981 bp cDNA containing the entire coding region of a human placental fructose-6-phosphate,2-kinase/fructose-2,6-bisphosphatase was determined. The sequence encodes 469 amino acids and, based on homology to the rat testis enzyme, appears to be the testis-type isozyme expressed in placenta. The enzyme was expressed in Escherichia coli BL21 (DE3) by using a T7 RNA polymerase-based expression system and purified to homogeneity. The expressed enzyme was bifunctional with specific activities of 75 and 80 mU/mg of kinase and phosphatase, respectively. Kinetic parameters of the expressed enzyme are similar to those of the rat testis enzyme.
测定了包含人胎盘果糖-6-磷酸-2-激酶/果糖-2,6-二磷酸酶完整编码区的1981 bp cDNA的核苷酸序列。该序列编码469个氨基酸，基于与大鼠睾丸酶的同源性，似乎是在胎盘中表达的睾丸型同工酶。通过使用基于T7 RNA聚合酶的表达系统在大肠杆菌BL21(DE3)中表达该酶，并纯化至同质。表达的酶具有双功能，激酶和磷酸酶的比活性分别为75和80 mU/mg。表达的酶的动力学参数与大鼠睾丸酶的相似。
