Lawrence Andrew D, Frank Stefanie, Newnham Sarah, Lee Matthew J, Brown Ian R, Xue Wei-Feng, Rowe Michelle L, Mulvihill Daniel P, Prentice Michael B, Howard Mark J, Warren Martin J
School of Biosciences, University of Kent, Giles Lane, Canterbury, Kent, CT2 7NJ, UK.
Department of Microbiology, University College Cork, Cork, Ireland.
ACS Synth Biol. 2014 Jul 18;3(7):454-465. doi: 10.1021/sb4001118. Epub 2014 Feb 24.
Targeting of proteins to bacterial microcompartments (BMCs) is mediated by an 18-amino-acid peptide sequence. Herein, we report the solution structure of the N-terminal targeting peptide (P18) of PduP, the aldehyde dehydrogenase associated with the 1,2-propanediol utilization metabolosome from Citrobacter freundii. The solution structure reveals the peptide to have a well-defined helical conformation along its whole length. Saturation transfer difference and transferred NOE NMR has highlighted the observed interaction surface on the peptide with its main interacting shell protein, PduK. By tagging both a pyruvate decarboxylase and an alcohol dehydrogenase with targeting peptides, it has been possible to direct these enzymes to empty BMCs in vivo and to generate an ethanol bioreactor. Not only are the purified, redesigned BMCs able to transform pyruvate into ethanol efficiently, but the strains containing the modified BMCs produce elevated levels of alcohol.
蛋白质靶向细菌微区室（BMCs）是由一个18个氨基酸的肽序列介导的。在此，我们报告了来自弗氏柠檬酸杆菌的与1,2-丙二醇利用代谢体相关的醛脱氢酶PduP的N端靶向肽（P18）的溶液结构。该溶液结构显示该肽在其全长上具有明确的螺旋构象。饱和转移差异和转移NOE NMR突出了该肽与其主要相互作用外壳蛋白PduK上观察到的相互作用表面。通过用靶向肽标记丙酮酸脱羧酶和醇脱氢酶，已能够在体内将这些酶引导至空的BMCs并生成乙醇生物反应器。不仅纯化的、重新设计的BMCs能够有效地将丙酮酸转化为乙醇，而且含有修饰的BMCs的菌株产生的酒精水平升高。
