Isolation and characterization of a pyrophosphate-dependent phosphofructokinase from Propionibacterium shermanii.
Isolation and characterization of a pyrophosphate-dependent phosphofructokinase from Propionibacterium shermanii.
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从谢尔曼丙酸杆菌中分离和表征焦磷酸依赖性磷酸果糖激酶。
DOI：
10.1016/s0021-9258(19)40727-8
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发表时间：
1975
期刊：
影响因子：
--
通讯作者：
Wood
中科院分区：
文献类型：
--
作者：
William;E.;O’BRIEN;Susanne;Bowien;Harland;G.;Wood
一种焦磷酸依赖性磷酸果糖激酶（焦磷酸; D-果糖-6-磷酸-1-磷酸转移酶）已从谢氏丙酸杆菌的提取物中纯化并表征。该酶催化磷酸从焦磷酸转移到果糖6-磷酸，以产生果糖-1，6-P2和磷酸。最初在溶组织内阿米巴中观察到这种独特的酶活性（Reeves，R.E.，南，D. J.，Blytt，H. G.，和Warren，L. G.（1974）J.Biol.Chem.249，7734-7741）。这是第三焦磷酸盐利用酶，这两个不同的生物体有共同之处。其他的是磷酸烯醇丙酮酸羧基转磷酸酶和丙酮酸磷酸二激酶。来自P. shermanii的PPi-磷酸果糖激酶特异性针对果糖-6-P和果糖-1，6-P2，不利用其他磷酸化糖。磷酸盐可以被砷酸盐取代。Km值为：磷酸盐，6.0 X 10（-4）M;果糖-1，6-P2，5.1 X 10（-5）M;焦磷酸盐，6.9 X 10（-5）M;果糖-6-P，1.0 X 10（-4）M。S20 w为5.1 S。天然酶的分子量为95，000。酶的十二烷基硫酸钠电泳显示迁移的单一条带，其Rf对应于48，000的分子量。P. shermanii的提取物具有比ATP-磷酸果糖激酶高约6倍的PPi-磷酸果糖激酶活性和比果糖二磷酸酶活性高15至20倍的PPi-磷酸果糖激酶活性。提出（a）当生物体在葡萄糖上生长时，PPi可以在果糖-1-6-P2的形成中取代ATP，和（B）当生物体在乳酸盐或甘油上生长时，在酵母发生期间，果糖-1，6-P2向果糖-6-P的转化可以通过磷酸解而不是水解发生。
A pyrophosphate-dependent phosphofructokinase (pyrophosphate; D-fructose-6-phosphate-1-phosphotransferase) has been purified and characterized from extracts of Propionibacterium shermanii. The enzyme catalyzes the transfer of phosphate from pyrophosphate to fructose 6-phosphate to yield fructose-1,6-P2 and phosphate. This unique enzymatic activity was observed initially in Entamoeba histolytica (Reeves, R.E., South, D.J., Blytt, H.G., and Warren, L. G. (1974) J. Biol. Chem. 249, 7734-7741). This is the third pyrophosphate-utilizing enzyme that these two diverse organisms have in common. The others are phosphoenolpyruvate carboxytransphosphorylase and pyruvate phosphate dikinase. The PPi-phosphofructokinase from P. shermanii is specific for fructose-6-P and fructose-1,6-P2, no other phosphorylated sugars were utilized. Phosphate could be replaced by arsenate. The Km values are: phosphate, 6.0 X 10(-4) M; fructose-1, 6-P2, 5.1 X 10(-5) M; pyrophosphate, 6.9 X 10(-5) M; and fructose-6-P, 1.0 X 10(-4) M. The S20w is 5.1 S. The molecular weight of the native enzyme is 95,000. Sodium dodecyl sulfate electrophoresis of the enzyme showed a single band migrating with an Rf corresponding to a molecular weight of 48,000. Extracts of P. shermanii have PPi-phosphofructokinase activity approximately 6 times greater than ATP-phosphofructokinase and 15 to 20 times greater than fructose diphosphatase activities. It is proposed that (a) PPi may replace ATP in the formation of fructose-1-6-P2 when the organism is grown on glucose and (b) when the organism is grown on lactate or glycerol the conversion of fructose-1,6-P2 to fructose-6-P during gluconeogenesis may occur by phosphorolysis rather than hydrolysis.