title: Engineering Microbes to Synthesize Plant Isoprenoids.
abstract: Humans constantly look for faster, more economical, and more sustainable ways to produce chemicals that originally harvested from nature. Over the past two decades, substantial progress has been made toward this goal by harnessing enzymes and cells as biocatalysts. For example, enzymes of slow-growing plants can be reconstituted in microbes, which empower them with the ability to produce useful plant metabolic compounds from sugars faster than plants. In this chapter, we provide protocols for producing isoprenoids - a large group of useful natural products - in microbes. It has been found that expression of genes encoding plant enzymes and selected endogenous genes must be delicately adjusted in microbes, otherwise isoprenoid production is negatively affected. Therefore, we focus on how to balance gene expression in Escherichia coli and use process engineering to increase its isoprenoid production. We also introduce our recent work on the use of microbial consortia and provide protocols for using yeast to help E. coli functionalize its isoprenoid product. Together, the methods and protocols provided here should be useful to researchers who aim to use microbes to synthesize novel isoprenoids.
authors.affiliation: National University of Singapore, Singapore, Singapore. Electronic address: kang.zhou@nus.edu.sg.
authors.forename: K
authors.initials: K
authors.lastname: Zhou
authors.affiliation: Massachusetts Institute of Technology, Cambridge, MA, United States.
authors.forename: S
authors.initials: S
authors.lastname: Edgar
authors.affiliation: Massachusetts Institute of Technology, Cambridge, MA, United States. Electronic address: gregstep@mit.edu.
authors.forename: G
authors.initials: G
authors.lastname: Stephanopoulos
mesh_terms: D019149:Bioreactors / Q000382:microbiology; D053898:Biosynthetic Pathways; D018920:Coculture Techniques / Q000379:methods; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D017343:Genes, Plant; D007218:Industrial Microbiology / Q000379:methods; D060847:Metabolic Engineering / Q000379:methods*; D010944:Plants / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D013729:Terpenes / Q000378:metabolism*; D014170:Transformation, Genetic
chemical_list: D013729:Terpenes
keywords: Escherichia coli; Isoprenoids; Metabolic engineering; Microbial consortium; Nonmevalonate pathway; Saccharomyces cerevisiae