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- Liu, Quanli, 1988, et al.
(author)
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Modular Pathway Rewiring of Yeast for Amino Acid Production
- 2018
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In: Methods in Enzymology. - : Elsevier. - 1557-7988 .- 0076-6879. ; 608, s. 417-439
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Book chapter (other academic/artistic)abstract
- Amino acids find various applications in biotechnology in view of their importance in the food, feed, pharmaceutical, and personal care industries as nutrients, additives, and drugs, respectively. For the large-scale production of amino acids, microbial cell factories are widely used and the development of amino acid-producing strains has mainly focused on prokaryotes Corynebacterium glutamicum and Escherichia coli. However, the eukaryote Saccharomyces cerevisiae is becoming an even more appealing microbial host for production of amino acids and derivatives because of its superior molecular and physiological features, such as amenable to genetic engineering and high tolerance to harsh conditions. To transform S. cerevisiae into an industrial amino acid production platform, the highly coordinated and multiple layers regulation in its amino acid metabolism should be relieved and reconstituted to optimize the metabolic flux toward synthesis of target products. This chapter describes principles, strategies, and applications of modular pathway rewiring in yeast using the engineering of L-ornithine metabolism as a paradigm. Additionally, detailed protocols for in vitro module construction and CRISPR/Cas-mediated pathway assembly are provided.
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| 2. |
- Campbell, Kate, 1987, et al.
(author)
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Biochemical principles enabling metabolic cooperativity and phenotypic heterogeneity at the single cell level
- 2018
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In: Current Opinion in Systems Biology. - : Elsevier BV. - 2452-3100. ; 8, s. 97-108
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Research review (peer-reviewed)abstract
- All biosynthetically active cells release metabolites, in part due to membrane leakage and cell lysis, but also in part due to overflow metabolism and ATP-dependent membrane export. At the same time, cells are adapted to sense and take up extracellular nutrients when available, to minimize the number of biochemical reactions that have to operate within a cell in parallel, and ultimately, to gain metabolic efficiency and biomass. Within colonies, biofilms or tissues, the co-occurrence of metabolite export and import enables the sharing of metabolites as well as metabolic specialization of single cells. In this review we discuss emerging biochemical concepts that give reasoning for why cells overproduce and release metabolites, and how these form the foundations for cooperative metabolite exchange activity between cells. We place particular emphasis on discussing the role of overflow metabolism in cells that exhibit either the Warburg or Crabtree effect. Furthermore, we discuss the profound physiological changes that cells undergo when their metabolism switches from metabolite synthesis to uptake, providing an explanation why metabolic specialization results in non-genotypic heterogeneity at the single cell level.
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