ModA

Background

Nitrogen fixation is a process in which nitrogen in the atmosphere is converted into ammonium (NH⁴⁺).No matter natural and synthetic, the process is essential for all forms of life because nitrogen is required to biosynthesize basic building blocks of plants, animals and other life forms, like nucleotides and amino acids.

All biological nitrogen fixation is done by way of nitrogenase, a metal-enzymes which contain iron, molybdenum, or vanadium. The enzyme is composed of the heterotetrameric MoFe protein that is transiently associated with the homodimeric Fe protein. Therefore, Molybdenum is an indispensable chemical element to nitrogen fixation and agriculture.

According to the Liebig's law of the minimum, which states that growth is controlled not by the total amount of resources available, but by the scarcest resource, molybdenum as a trace mineral, is identified as limiting the growth of peanuts in large field of China. So molybdenum powder is commonly used as a fertilizer in agriculture. And our project aims to establish an E.Coli strain that is able to capture Mo in the medium, directional deliver itself to root system and kill itself eventually.

ModA

The genes encoding a molybdate transport system (mod) have been isolated and sequenced from Escherichia coli[2–5] Four genes modABCD, organized in an operon, share strong structural similarity and sequence homology with genes encoding binding -protein dependent solute(or ABC-type) transport systems [2–5]. The first gene, modA, encodes a periplasmic binding protein. ModB and ModC have sequence homology with integral membrane and cytoplasmic ATP hydrolysis components of ABC transport systems, respectively. ModD does not show sequence similarity to any gene product in current databases and its role in transport is unclear. Two more genes, modEF, are also involved in molybdate transport. The modE gene encodes a regulatory protein that effects molybdenum-dependent repression at the modA promoter [6–8]. The exact role of modF is not yet clear [6].

The mod operon encodes a very high affinity(Km=15 nM) molybdate transport system. The low K values agree well with previous results obtained with the enteric diazotroph K. pneumoniae ([1]; Km=20 nM, J. Imperial, unpublished results). and adequately explain the high molybdenum scavenging capacity observed with most bacteria[11].

ModA, the periplasmic binding component of the mod system. had a K for molybdate of 3–5 mM. This is one of the highest substrate Kd values reported for any periplasmic binding protein [9,10] and contrasts with the low Kd values for molybdate uptake The Structure of modA[14]

How to bind molybdenum[15]

The Story of E.coli Prometheus

BNU-China