Metabolic pathway engineering using the central signal processor PII

Microbial Cell Factories

Research

Mark Stahl¹  Alicia Engelbrecht²  Iris Maldener²  Björn Watzer²  Waldemar Hauf²  Karl Forchhammer² 

[1] Central Facilities, Analytics, ZMBP, Eberhard-Karls-Universität Tübingen, Auf der Morgenstelle 32, 72076, Tübingen, Germany;Interfaculty Institute of Microbiology and Infection Medicine Tübingen, Eberhard-Karls-Universität Tübingen, Auf der Morgenstelle 28, 72076, Tübingen, Germany;

关键词: Cyanophycin;    Cyanobacteria;    -Arginine;    P protein;   

DOI  :  10.1186/s12934-015-0384-4

 received in 2015-07-28, accepted in 2015-11-06,  发布年份 2015

来源: Springer

【 摘 要 】

BackgroundPII signal processor proteins are wide spread in prokaryotes and plants where they control a multitude of anabolic reactions. Efficient overproduction of metabolites requires relaxing the tight cellular control circuits. Here we demonstrate that a single point mutation in the PII signaling protein from the cyanobacterium Synechocystis sp. PCC 6803 is sufficient to unlock the arginine pathway causing over accumulation of the biopolymer cyanophycin (multi-l-arginyl-poly-l-aspartate). This product is of biotechnological interest as a source of amino acids and polyaspartic acid. This work exemplifies a novel approach of pathway engineering by designing custom-tailored PII signaling proteins. Here, the engineered Synechocystis sp. PCC6803 strain with a PII-I86N mutation over-accumulated arginine through constitutive activation of the key enzyme N-acetylglutamate kinase (NAGK).ResultsIn the engineered strain BW86, in vivo NAGK activity was strongly increased and led to a more than tenfold higher arginine content than in the wild-type. As a consequence, strain BW86 accumulated up to 57 % cyanophycin per cell dry mass under the tested conditions, which is the highest yield of cyanophycin reported to date. Strain BW86 produced cyanophycin in a molecular mass range of 25 to >100 kDa; the wild-type produced the polymer in a range of 30 to >100 kDa.ConclusionsThe high yield and high molecular mass of cyanophycin produced by strain BW86 along with the low nutrient requirements of cyanobacteria make it a promising means for the biotechnological production of cyanophycin. This study furthermore demonstrates the feasibility of metabolic pathway engineering using the PII signaling protein, which occurs in numerous bacterial species.

【 授权许可】

CC BY   
© Watzer et al. 2015