Pathway construction

Steinbuchel, A. and Lutke-Eversloh, T. 2003. Metabolic engineering and pathway construction for biotechnological production of relevant polyhydroxyalkanoates in microorganisms. Biochemical Engineering Journal 16 81-96. 

These alterations stand as important examples of the flexibility of pathway construction when enough is known about the individual components of these pathways. 

In this recombinant pathway construct, under conditions of 50% atmospheric Oz, the pentachloroethane is reduced to TCE, which is subsequently oxidized by the constitutively expressed Tod. 

Non-catabolic genes for catabolic pathway constructions Bacterial hemoglobin... 

Despite these field trials, there has never been a release of a geneticaly engineered microorganism for bioremediation purposes. One should therefore bear in mind, when considering pathway construction as a route to bioremediation, that the constructed organism cannot carry out its programmed function without interacting with its environment. That we can carry out fairly complex alterations of the... 

Kwon SK, Lee MY, Ku B, Sherman DH, Dordick JS. High-throughput, microarray-based synthesis of natural product analogues via in vitro metabolic pathway construction. ACS Chem. Biol. 2007 2 419-425. 

How are specific pathways constructed from individual reactions A pathway must satisfy minimally two criteria (1) the individual reactions must be specific and (2) the entire set of reactions that constitute the pathway must be thermodynamically favored. A reaction that is specific will yield only one particular product or set of products from its reactants. As discussed in Chapter 8. a function of enzymes is to provide this specificity. The thermodynamics of metabolism is most readily approached in terms of free energy, which was discussed in Sections 1.3.3. 8.2.1, and 8.2.2. 

The procedure identify-nondissipative-pathways constructs the set of enabling conditions that lead to a top-level event. It takes as its input the variable-influence pathways, which it obtains from the procedure, construct-variable-influence-pathways. TLE variables are then identified, and each variable contained in the set is traced to identify potentially feasible roots for causing the TLE. When an achievable root is identified (i.e., an input disturbance, controlled or uncontrolled, which can enable the top-level event), it is collected into root causes and returned. The algorithm used by this procedure is shown below ... 

Miller CA 3rd. 1999. A human aryl hydrocarbon receptor signaling pathway constructed in yeast displays additive responses to ligand mixtures. Toxicol. Appl. Pharmacol. 160 297-303... 

Elimination of a branched pathway - construction of a LDH-A mutant E.coli... 

Steinbuchel, A., Lutke-Eversloh, T. (2003). Metabolic Engineering and Pathway Construction for Biotechnological Production of Relevant Poly Hydroxyalkanoates in Microorganisms. Biochem. Eng. J., 16, 81-96. 

Recent technical advances in strain/pathway construction have moved the limiting step from strain construction to screening. Those advances are, among others, affordable gene synthesis, synthetic construction of gene libraries (Collier et al., 2009 Bott et al., 2012), and novel techniques for DNA assembly (e.g., Engler et al., 2008 Gibson et al., 2009). 

Ui S, Takusagawa Y, Sato T, Ohtsuki T, Mimura A, Ohkuma M, Kudo T. (2004). Production of L-2, 3-butanediol by a new pathway constructed in Escherichia coli. Lett Appl Microbiol, 39, 533-537. 

Li S, Wen J, Jia X. (2011). Engineering Bacillus subtilis for isobutanol production by heterologous Ehrlich pathway construction and the biosynthetic 2-ketoisovalerate precursor pathway overexpression. Appl Microbiol Biotechnol, 91, 577-589. 

Carter OA, Peters RJ, Croteau R (2003) Monoter-pene biosynthesis pathway construction in Escherichia coli. Phytochemistry 64 425-433... 

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