OXIDATION OF D-LACTATE AND L-LACTATE BY NEISSERIA-MENINGITIDIS - PURIFICATION AND CLONING OF MENINGOCOCCAL D-LACTATE DEHYDROGENASE
OXIDATION OF D-LACTATE AND L-LACTATE BY NEISSERIA-MENINGITIDIS - PURIFICATION AND CLONING OF MENINGOCOCCAL D-LACTATE DEHYDROGENASE
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DOI：
10.1128/jb.175.20.6382-6391.1993
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发表时间：
1993-10-01
影响因子：
3.2
通讯作者：
GOTSCHLICH, EC
中科院分区：
文献类型：
--
作者：
ERWIN, AL;GOTSCHLICH, EC
发现脑膜炎奈瑟菌含有至少两种乳酸氧化酶。其中一种从原生质球膜中纯化 460 倍，发现主要对 D-乳酸具有特异性，对 L-乳酸具有低亲和力活性。克隆了该酶 (dld) 的基因，并通过插入失活该基因构建了 dld 突变体。该突变体无法在 D-乳酸上生长，但保留了在 L-乳酸上生长的能力，这为第二种对 L-乳酸具有特异性的乳酸氧化酶提供了证据。在 dld+ 和 dld 突变菌株的完整细菌中检测到高亲和力 L-乳酸氧化活性。这种 L-乳酸氧化活性也出现在超声处理的细菌中，但在洗涤剂溶解或原生质球膜制备时显着降低。
Neisseria meningitidis was found to contain at least two lactate-oxidizing enzymes. One of these was purified 460-fold from spheroplast membranes and found to be specific primarily for D-lactate, with low-affinity activity for L-lactate. The gene for this enzyme (dld) was cloned, and a dld mutant was constructed by insertional inactivation of the gene. The mutant was unable to grow on D-lactate but retained the ability to grow on L-lactate, providing evidence for a second lactate-oxidizing enzyme with specificity for L-lactate. High-affinity L-lactate-oxidizing activity was detected in intact bacteria of both the dld+ and dld mutant strains. This L-lactate-oxidizing activity was also seen in sonicated bacteria but was reduced substantially on detergent solubilization or on preparation of spheroplast membranes.