Kümmel A, Behrens G, Gottschalk G
Arch Microbiol. 1975;102(2):111-6. doi: 10.1007/BF00428354.
Citrate lyase (EC 4.1.3.6) was purified 38-fold from cell-free extracts of Streptococcus diacetilactis. The enzyme was homogeneous in analytical ultracentrifugation and polyacrylamide gel electrophoresis. The final enzyme preparation contained acetate: HS-citrate lyase ligase--an acetylating enzyme which converts inactive HS-citrate lyase into enzymatically active acetyl-S-citrate lyase. This enzyme activity was purified 25-fold over the crude extract and seemed to be associated with citrate lyase. Partially purified citrate lyase from Leuconostoc citrovorum contained also its acetylating enzyme. Purified citrate lyases from Klebsiella aerogenes and Rhodopseudomonas gelatinosa were devoid of acetylating enzyme activity. The HS-form of citrate lyase from S. diacetilactis was completely acetylated and hence activated by incubation with ATP and acetate for 25 min at 25 degrees C. The enzyme did not acetylate the HS-lyases from R. gelatinosa and K. aerogenes. In contrast to the citrate lyases from R. gelatinosa and K. aerogenes the enzymes from S. diacetilactis and L. citrovorum showed only a very weak reaction inactivation. It is assumed that this is due to the association of the acetylating enzymes with these lyases.
从双乙酰乳酸链球菌的无细胞提取物中纯化出了38倍的柠檬酸裂解酶（EC 4.1.3.6）。该酶在分析超速离心和聚丙烯酰胺凝胶电泳中均呈均一状态。最终的酶制剂含有乙酸：HS-柠檬酸裂解酶连接酶——一种乙酰化酶，它将无活性的HS-柠檬酸裂解酶转化为具有酶活性的乙酰-S-柠檬酸裂解酶。这种酶活性比粗提取物纯化了25倍，且似乎与柠檬酸裂解酶相关。来自嗜柠檬酸明串珠菌的部分纯化的柠檬酸裂解酶也含有其乙酰化酶。来自产气克雷伯菌和胶状红假单胞菌的纯化柠檬酸裂解酶没有乙酰化酶活性。来自双乙酰乳酸链球菌的HS形式的柠檬酸裂解酶通过在25℃下与ATP和乙酸孵育25分钟而完全乙酰化并因此被激活。该酶不会使来自胶状红假单胞菌和气克雷伯菌的HS-裂解酶乙酰化。与来自胶状红假单胞菌和气克雷伯菌的柠檬酸裂解酶不同，来自双乙酰乳酸链球菌和嗜柠檬酸明串珠菌的酶仅表现出非常微弱的反应失活。据推测，这是由于乙酰化酶与这些裂解酶的结合所致。
