Modular Pathway Rewiring of Yeast for Amino Acid Production
Book chapter, 2018

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.

Saccharomyces cerevisiae

Modular pathway engineering

Amino acids

Metabolic flux

L-Ornithine

Author

Quanli Liu

Novo Nordisk

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Tao Yu

Novo Nordisk

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Kate Campbell

Novo Nordisk

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Jens Christian Froslev Nielsen

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Novo Nordisk

Yun Chen

Novo Nordisk

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Methods in Enzymology

417-439

Subject Categories

Microbiology

Biocatalysis and Enzyme Technology

Organic Chemistry

DOI

10.1016/bs.mie.2018.06.009

PubMed

30173772

More information

Latest update

3/19/2019