Tanaka Yukinori, Kasahara Ken, Izawa Masumi, Ochi Kozo
a Department of Life Sciences , Hiroshima Institute of Technology , Hiroshima , Japan.
b Chitose Laboratory Corp. , Biotechnology Research Center , Miyamae-ku , Japan.
Biosci Biotechnol Biochem. 2017 Aug;81(8):1636-1641. doi: 10.1080/09168451.2017.1329619. Epub 2017 May 22.
Ribosome engineering has been widely utilized for strain improvement, especially for the activation of bacterial secondary metabolism. This study assessed ribosome engineering technology to modulate primary metabolism, taking vitamin B12 production as a representative example. The introduction into Propionibacterium shermanii of mutations conferring resistance to rifampicin, gentamicin, and erythromycin, respectively, increased per cell production (μg/L/OD) of vitamin B12 5.2-fold, although net production (μg/L) was unchanged, as the cell mass of the mutants was reduced. Real-time qPCR analysis demonstrated that the genes involved in vitamin B12 fermentation by P. shermanii were activated at the transcriptional level in the drug-resistant mutants, providing a mechanism for the higher yields of vitamin B12 by the mutants. These results demonstrate the efficacy of ribosome engineering for the production of not only secondary metabolites but of industrially important primary metabolites.
核糖体工程已被广泛用于菌株改良,特别是用于激活细菌的次级代谢。本研究以维生素B12的生产为例,评估了核糖体工程技术对初级代谢的调节作用。分别向谢氏丙酸杆菌引入赋予对利福平、庆大霉素和红霉素抗性的突变,维生素B12的每细胞产量(μg/L/OD)提高了5.2倍,尽管净产量(μg/L)没有变化,因为突变体的细胞量减少了。实时定量PCR分析表明,谢氏丙酸杆菌中参与维生素B12发酵的基因在耐药突变体中在转录水平上被激活,这为突变体中维生素B12产量更高提供了一种机制。这些结果证明了核糖体工程不仅对次级代谢产物的生产有效,而且对工业上重要的初级代谢产物的生产也有效。