Inhibition of AckA and Pta Genes Using Two Specific Antisense RNAs Reduced Acetate Accumulation in Batch Fermentation of E. coli BL21 (DE3)

Down Regulation of ackA-pta Pathway in Escherichia coli BL21 (DE3): A Step Toward Optimized Recombinant Protein Expression System

BACKGROUND One of the most important problems in production of recombinant protein is to attain over-expression of the target gene and high cell density. In such conditions, the secondary metabolites of bacteria become toxic for the medium and cause cells to die. One of these aforementioned metabolites is acetate, which enormously accumulated in the medium, so that both cell and protein yields ...

Effects of ackA, pta and poxB inhibition by antisense RNA on acetate excretion and recombinant beta interferon expression in Escherichia coli

Introduction: Escherichia coli (E.coli) is one of the most widely used hosts for the production of recombinant proteins. The main problem in getting high product yields and productivity is the accumulation of acetic acid (acetate) as an unwanted metabolic by-product. In this study, an antisense-based strategy as a metabolic engineering approach was employed to hamper the acetate excretion probl...

down-regulation of acetate pathway through antisense rna against acka and pta in escherichia coli: improved beta interferon production

down regulation of acka-pta pathway in escherichia coli bl21 (de3): a step toward optimized recombinant protein expression system

materials and methods: in order to achieve this, recombinant plasmid harboring antisense sequences targeting both of pta and acka was assembled, after transfecting to the cells, its effects on the cell growth and acetate accumulation in the minimal media was assessed and compared with the control, the plasmid without antisense cassette, in presence and absence of iptg in escherichia coli bl21 (...

Acetate fluxes in Escherichia coli are determined by the thermodynamic control of the Pta-AckA pathway

Escherichia coli excretes acetate upon growth on fermentable sugars, but the regulation of this production remains elusive. Acetate excretion on excess glucose is thought to be an irreversible process. However, dynamic 13C-metabolic flux analysis revealed a strong bidirectional exchange of acetate between E. coli and its environment. The Pta-AckA pathway was found to be central for both flux di...

Batch production of a silk-elastin-like protein in E. coli BL21(DE3): key parameters for optimisation

BACKGROUND Silk-elastin-like proteins (SELPs) combining the physicochemical and biological properties of silk and elastin have a high potential for use in the pharmaceutical, regenerative medicine and materials fields. Their development for use is however restrained by their production levels. Here we describe the batch production optimisation for a novel recently described SELP in the pET-E. c...