Towards the engineering of in vitro systems

Abstract

Synthetic biology aims at rationally implementing biological systems from scratch. Given the complexity of living systems and our current lack of understanding of many aspects of living cells, this is a major undertaking. The design of in vitro systems can be considerably easier, because they can consist of fewer constituents, are quasi time invariant, their parameter space can be better accessed and they can be much more easily perturbed and then analysed chemically and mathematically. However, even for simplified in vitro systems, following a comprehensively rational design procedure is still difficult. When looking at a comparatively simple system, such as a medium-sized enzymatic reaction network as it is represented by glycolysis, major issues such as a lack of comprehensive enzyme kinetics and of suitable knowledge on crucial design parameters remain. Nevertheless, in vitro systems are very suitable to overcome these obstacles and therefore well placed to act as a stepping stone to engineering living systems.

A synthetic biochemistry molecular purge valve module that maintains redox balance

“…Microbial systems are often hampered by a variety of technical challenges that make it hard to achieve cost competitiveness, including poor yields owing to competing pathways; low productivity caused by slow growth rates or difficulties in pathway optimization; contaminating microbial growth; product toxicity; and expensive product isolation 2,3 .…”

Cell-free Biosystems in the Production of Electricity and Bioenergy

“…Two ways for constructing cell-free biosystems are whole crude cell extracts and purified enzymes. The former is easily prepared but most enzyme components may not last long and the existing undesired pathways may cause side reactions [4,[104][105][106]. The latter requires more labor and cost for enzyme preparation but it has better access control and easy optimization of numerous building blocks with different characteristics.…”

Hazards, Risks, and Low Hazard Development Paths of Synthetic Biology

“…But, at least partially, mechanisms of self-repair can already be compensated by optimized reaction conditions, optimized amino acid sequences to minimize covalent interactions or even the additional implementation of repair mechanisms (Hold and Panke 2009).…”