Hirasawa T, Wachi M, Nagai K
Department of Bioengineering, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama, Japan.
BMC Biotechnol. 2001;1:9. doi: 10.1186/1472-6750-1-9. Epub 2001 Oct 16.
A non-pathogenic species of coryneform bacteria, Corynebacterium glutamicum, was originally isolated as an L-glutamate producing bacterium and is now used for fermentative production of various amino acids. A mutation in the C. glutamicum ltsA gene caused susceptibility to lysozyme, temperature-sensitive growth, and L-glutamate production.
The characteristics of eight lysozyme-sensitive mutants which had been isolated after N-methyl-N'-nitro-N-nitrosoguanidine mutagenesis were examined. Complementation analysis with the cloned wild-type ltsA gene and DNA sequencing of the ItsA region revealed that four mutants had a mutation in the ltsA gene. Among them, two mutants showed temperature-sensitive growth and overproduced L-glutamate at higher temperatures, as well as the previously reported ltsA mutant. Other two showed temperature-resistant growth: one missense mutant produced L-glutamate to some extent but the other nonsense mutant did not. These two mutants remained temperature-resistant in spite of introduction of ltsA::kan mutation that causes temperature-sensitive growth in the wild-type background.
These results indicate that a defect caused by the ltsA mutations is responsible for temperature-sensitive growth and L-glutamate overproduction by C. glutamicum. The two temperature-resistant mutants seem to carry suppressor mutations that rendered cells temperature-resistance and abolished L-glutamate overproduction.
棒状杆菌属的一种非致病性细菌,谷氨酸棒杆菌,最初作为一种产生L-谷氨酸的细菌被分离出来,现在用于多种氨基酸的发酵生产。谷氨酸棒杆菌ltsA基因的突变导致对溶菌酶敏感、温度敏感生长以及L-谷氨酸的产生。
对经N-甲基-N'-硝基-N-亚硝基胍诱变后分离得到的八个溶菌酶敏感突变体的特性进行了研究。用克隆的野生型ltsA基因进行互补分析以及对ltsA区域进行DNA测序表明,四个突变体在ltsA基因中有突变。其中,两个突变体表现出温度敏感生长,并且在较高温度下过量产生L-谷氨酸,与之前报道的ltsA突变体一样。另外两个表现出温度抗性生长:一个错义突变体在一定程度上产生L-谷氨酸,但另一个无义突变体则不产生。尽管引入了ltsA::kan突变(该突变在野生型背景下会导致温度敏感生长),这两个突变体仍然具有温度抗性。
这些结果表明,ltsA突变引起的缺陷是谷氨酸棒杆菌温度敏感生长和L-谷氨酸过量产生的原因。这两个温度抗性突变体似乎携带抑制突变,使细胞具有温度抗性并消除了L-谷氨酸的过量产生。