The metabolic potential of Escherichia coli BL21 in defined and rich medium
نویسندگان
چکیده
BACKGROUND The proteome reflects the available cellular machinery to deal with nutrients and environmental challenges. The most common E. coli strain BL21 growing in different, commonly employed media was evaluated using a detailed quantitative proteome analysis. RESULTS The presence of preformed biomass precursor molecules in rich media such as Luria Bertani supported rapid growth concomitant to acetate formation and apparently unbalanced abundances of central metabolic pathway enzymes, e.g. high levels of lower glycolytic pathway enzymes as well as pyruvate dehydrogenase, and low levels of TCA cycle and high levels of the acetate forming enzymes Pta and AckA. The proteome of cells growing exponentially in glucose-supplemented mineral salt medium was dominated by enzymes of amino acid synthesis pathways, contained more balanced abundances of central metabolic pathway enzymes, and a lower portion of ribosomal and other translational proteins. Entry into stationary phase led to a reconstruction of the bacterial proteome by increasing e.g. the portion of proteins required for scavenging rare nutrients and general cell protection. This proteomic reconstruction during entry into stationary phase was more noticeable in cells growing in rich medium as they have a greater reservoir of recyclable proteins from the translational machinery. CONCLUSIONS The proteomic comparison of cells growing exponentially in different media reflected the antagonistic and competitive regulation of central metabolic pathways through the global transcriptional regulators Cra, Crp, and ArcA. For example, the proteome of cells growing exponentially in rich medium was consistent with a dominating role of phosphorylated ArcA most likely a result from limitations in reoxidizing reduced quinones in the respiratory chain under these growth conditions. The proteomic alterations of exponentially growing cells into stationary phase cells were consistent with stringent-like and stationary phase responses and a dominating control through DksA-ppGpp and RpoS.
منابع مشابه
Determination of the optimal conditions of cloning Aerolysin gene from the common carp pathogen Aeromonas hydrophila in Escherichia coli BL21
Aeromonas hydrophila is a gram-negative bacterium which associated with gastrointestinal diseases and septicaemia. This pathogenic bacterium has several virulence factors ranging from pili to the excreted protein which called (Aerolysin) with minor and major effects, respectively. Additionally, Aeromonas hydrophila is a widely distributed bacterium that commonly causes ulcers in cyprinid fish s...
متن کاملProduction of the short peptide surfactant DAMP4 from glucose or sucrose in high cell density cultures of Escherichia coli BL21(DE3)
BACKGROUND Peptides are increasingly used in industry as highly functional materials. Bacterial production of recombinant peptides has the potential to provide large amounts of renewable and low cost peptides, however, achieving high product titers from Chemically Defined Media (CDM) supplemented with simple sugars remains challenging. RESULTS In this work, the short peptide surfactant, DAMP4...
متن کاملActivation of the Glutamic Acid-Dependent Acid Resistance System in Escherichia coli BL21(DE3) Leads to Increase of the Fatty Acid Biotransformation Activity
The biosynthesis of carboxylic acids including fatty acids from biomass is central in envisaged biorefinery concepts. The productivities are often, however, low due to product toxicity that hamper whole-cell biocatalyst performance. Here, we have investigated factors that influence the tolerance of Escherichia coli to medium chain carboxylic acid (i.e., n-heptanoic acid)-induced stress. The met...
متن کاملExpression of Recombinant Protein B Subunit Pili from Vibrio Cholera
Background & Aims: Vibrio cholerae is a gram-negative bacterial pathogen that causes cholera disease. Following ingestion by a host and entry into the upper intestine, V. cholera colonizes and begins to emit enterotoxin. One of the most pathogenic factors of Vibrio cholera is toxin-coregulated pili (TCP). ToxinCoregulated pili is as the primary factor requiered for the colonization and insisten...
متن کاملTransformation and gene expression study of recombinant fish GnRH in E. coli BL21 in order to produce recombinant hormone
Gonadotropin releasing hormone (GnRH) is a neuropeptide known to regulate reproduction in vertebrates. Different analogues of synthetic GnRH are used to induce final sexual maturation in fish breeders. The purpose of this research was to evaluate the expression of recombinant GnRH (rGnRH) in Escherichia coli BL21 to produce recombinant hormone. In the present research, the sequence of DNA relat...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
دوره 13 شماره
صفحات -
تاریخ انتشار 2014