Inverse metabolic engineering: A strategy for directed genetic engineering of useful phenotypes

TLDR

Classical metabolic engineering, which targets rate‑determining steps and overexpresses enzymes, has had limited practical success because other limiting steps, regulatory counter‑balancing, and unknown coupled pathways often confound the direct approach. The authors codify inverse metabolic engineering and illustrate its application with several examples. Inverse metabolic engineering involves first defining a desired phenotype, then identifying the genetic or environmental factors that produce it, and finally transferring that phenotype to another strain or organism through directed manipulation. This paradigm has been successfully applied to eliminate growth‑factor requirements in mammalian cell culture and to increase the energetic efficiency of microaerobic bacterial respiration. © 2002 Wiley Periodicals, Inc., Biotechnol Bioeng 79: 568–579.

Abstract

Abstract The classical method of metabolic engineering, identifying a rate‐determining step in a pathway and alleviating the bottleneck by enzyme overexpression, has motivated much research but has enjoyed only limited practical success. Intervention of other limiting steps, of counter‐balancing regulation, and of unknown coupled pathways often confounds this direct approach. Here the concept of inverse metabolic engineering is codified and its application is illustrated with several examples. Inverse metabolic engineering means the elucidation of a metabolic engineering strategy by: first, identifying, constructing, or calculating a desired phenotype; second, determining the genetic or the particular environmental factors conferring that phenotype; and third, endowing that phenotype on another strain or organism by directed genetic or environmental manipulation. This paradigm has been successfully applied in several contexts, including elimination of growth factor requirements in mammalian cell culture and increasing the energetic efficiency of microaerobic bacterial respiration. © 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 79: 568–579, 2002.