Kikuchi Yoshimi, Itaya Hiroshi, Date Masayo, Matsui Kazuhiko, Wu Long-Fei
Institute of Life Sciences, Ajinomoto Co., Inc., Kawasaki, Japan.
Appl Environ Microbiol. 2009 Feb;75(3):603-7. doi: 10.1128/AEM.01874-08. Epub 2008 Dec 12.
The twin-arginine translocation (Tat) pathway in Corynebacterium glutamicum has been described previously. The minimal functional Tat system in C. glutamicum required TatA and TatC but did not require TatB, although this component was required for maximal efficiency of Tat-dependent secretion. We previously demonstrated that Chryseobacterium proteolyticum pro-protein glutaminase (pro-PG) and Streptomyces mobaraensis pro-transglutaminase (pro-TG) could be secreted via the Tat pathway in C. glutamicum. Here we report that the amounts of pro-PG secreted were more than threefold larger when TatC or TatAC was overexpressed, and there was a further threefold increase when TatABC was overexpressed. These results show that the amount of TatC protein is the first bottleneck and the amount of TatB protein is the second bottleneck in Tat-dependent protein secretion in C. glutamicum. In addition, the amount of pro-TG that accumulated via the Tat pathway when TatABC was overexpressed with the TorA signal peptide in C. glutamicum was larger than the amount that accumulated via the Sec pathway. We concluded that TatABC overexpression improves Tat-dependent pro-PG and pro-TG secretion in C. glutamicum.
谷氨酸棒杆菌中的双精氨酸转运(Tat)途径此前已有描述。谷氨酸棒杆菌中最小的功能性Tat系统需要TatA和TatC,但不需要TatB,尽管该组分对于Tat依赖性分泌的最大效率是必需的。我们先前证明,解蛋白金黄杆菌的前体蛋白谷氨酰胺酶(pro-PG)和茂原链霉菌的前体转谷氨酰胺酶(pro-TG)可以通过谷氨酸棒杆菌中的Tat途径分泌。在此我们报告,当TatC或TatAC过表达时,分泌的pro-PG量增加了三倍以上,而当TatABC过表达时,又增加了三倍。这些结果表明,TatC蛋白的量是谷氨酸棒杆菌中Tat依赖性蛋白质分泌的第一个瓶颈,而TatB蛋白的量是第二个瓶颈。此外,当在谷氨酸棒杆菌中用TorA信号肽过表达TatABC时,通过Tat途径积累的pro-TG量大于通过Sec途径积累的量。我们得出结论,TatABC过表达可改善谷氨酸棒杆菌中Tat依赖性pro-PG和pro-TG的分泌。