ACKA E.Coli
Description
The ACKA is fused to a 23 amino acid His-Tag at N-terminus and purified by proprietary chromatographic techniques.
Product Specs
Product Science Overview
Acetate kinase catalyzes the reversible transfer of a phosphate group from ATP to acetate, forming acetyl phosphate and ADP. This reaction is a key step in the metabolic pathway that allows E. coli to utilize acetate as a carbon source. The enzyme is composed of two identical subunits, each with a molecular weight of approximately 43 kDa. The active site of acetate kinase binds both ATP and acetate, facilitating the transfer of the phosphate group.
In E. coli, acetate kinase is part of the central metabolic pathway that connects glycolysis, the tricarboxylic acid (TCA) cycle, and the glyoxylate shunt. During aerobic growth on glucose, E. coli can produce acetate as a by-product through a process known as overflow metabolism. This occurs when the rate of glucose uptake exceeds the capacity of the TCA cycle, leading to the accumulation of acetyl-CoA, which is then converted to acetate by the ackA-pta pathway .
The recombinant production of acetate kinase involves cloning the gene encoding the enzyme into a suitable expression vector, which is then introduced into an E. coli host strain. The recombinant E. coli cells are cultured under conditions that induce the expression of the acetate kinase gene, leading to the production of large quantities of the enzyme. This recombinant enzyme can be purified using standard biochemical techniques, such as affinity chromatography.
Recombinant acetate kinase from E. coli has several applications in research and industry:
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Metabolic Engineering: Acetate kinase is used in metabolic engineering to enhance the production of various biochemicals. For example, overexpression of the ackA-pta pathway can improve the assimilation of acetate and increase the yield of products such as polyhydroxyalkanoates (PHAs), which are biodegradable plastics .
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Biochemical Studies: The enzyme is used to study the regulation of acetate metabolism and its impact on cellular physiology. Understanding the role of acetate kinase in E. coli can provide insights into the mechanisms of overflow metabolism and the strategies to minimize acetate accumulation during high-density fermentations .
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Industrial Biotechnology: Acetate kinase is employed in the production of recombinant proteins and other bioproducts. By optimizing the expression and activity of acetate kinase, it is possible to reduce the inhibitory effects of acetate on cell growth and protein production, thereby improving the overall efficiency of bioprocesses .
One of the main challenges in using acetate kinase in industrial applications is the inhibitory effect of acetate on E. coli growth. High concentrations of acetate can lead to reduced cell density and lower yields of recombinant proteins. To address this issue, researchers are exploring various strategies, such as metabolic engineering and process optimization, to enhance acetate tolerance and minimize its accumulation .
Future research on acetate kinase may focus on:
- Engineering more efficient enzyme variants: By modifying the enzyme’s structure, it may be possible to improve its catalytic efficiency and stability under industrial conditions.
- Developing novel expression systems: Using alternative host organisms or expression vectors could enhance the production and activity of recombinant acetate kinase.
- Integrating acetate kinase into synthetic biology circuits: This could enable the design of more robust and efficient microbial cell factories for the production of biofuels, chemicals, and materials.
In conclusion, acetate kinase from E. coli is a versatile enzyme with significant potential in various biotechnological applications. Continued research and development in this area will likely lead to new and improved methods for harnessing the power of this enzyme in industrial processes.
