AUTHOR=Hung Chun-Hsien , Kanehara Kazue , Nakamura Yuki 

TITLE=In vivo Reconstitution of Algal Triacylglycerol Production in Saccharomyces cerevisiae

JOURNAL=Frontiers in Microbiology

VOLUME=7

YEAR=2016

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2016.00070

DOI=10.3389/fmicb.2016.00070

ISSN=1664-302X

ABSTRACT=<p>The current fascination with algal biofuel production stems from a high lipid biosynthetic capacity and little conflict with land plant cultivation. However, the mechanisms which enable algae to accumulate massive oil remain elusive. An enzyme for triacylglycerol (TAG) biosynthesis in <italic>Chlamydomonas reinhardtii</italic>, CrDGTT2, can produce a large amount of TAG when expressed in yeast or higher plants, suggesting a unique ability of CrDGTT2 to enhance oil production in a heterologous system. Here, we performed metabolic engineering in <italic>Saccharomyces cerevisiae</italic> by taking advantage of CrDGTT2. We suppressed membrane phospholipid biosynthesis at the log phase by mutating <italic>OPI3</italic>, enhanced TAG biosynthetic pathway at the stationary phase by overexpressing <italic>PAH1</italic> and <italic>CrDGTT2</italic>, and suppressed TAG hydrolysis on growth resumption from the stationary phase by knocking out <italic>DGK1</italic>. The resulting engineered yeast cells accumulated about 70-fold of TAG compared with wild type cells. Moreover, TAG production was sustainable. Our results demonstrated the enhanced and sustainable TAG production in the yeast synthetic platform.</p>