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Mutations in Escherichia coli polyphosphate kinase that lead to dramatically increased in vivo polyphosphate levels

J Bacteriol. 2018-02; 
Rudat AK, Pokhrel A, Green TJ, Gray MJ
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Bacterial Expression System The ppgK (all1371) coding sequence (717 bp) from Anabaena sp. strain PCC 7120 (17) was codon optimized for expression in E. coli (GenScript), yielding a ppgKopt. allele encoding the wild-type PPGK amino acid sequence in plasmid pPPGK1. Get A Quote

摘要

Bacteria synthesize inorganic polyphosphate (polyP) in response to a wide variety of stresses, and production of polyP is essential for stress response and survival in many important pathogens and bacteria used in biotechnological processes. However, surprisingly little is known about the molecular mechanisms that control polyP synthesis. We have therefore developed a novel genetic screen that specifically links growth of Escherichia coli to polyP synthesis, allowing us to isolate mutations leading to enhanced polyP production. Using this system, we have identified mutations in the polyP-synthesizing enzyme polyP kinase (PPK) that lead to dramatic increases in in vivo polyP synthesis but do not substantially af... More

关键词

bacterial genetics; enzyme engineering; inorganic polyphosphate; mutagenesis; stress response