Engineering 3-Hydroxypropionic Acid Production from Glucose in Yarrowia lipolytica through Malonyl-CoA Pathway

Abstract

3-Hydroxypropionic acid (3-HP) is an important intermediate compound in the chemical industry. Green and environmentally friendly microbial synthesis methods are becoming increasingly popular in a range of industries. Compared to other chassis cells, <i>Yarrowia lipolytica</i> possesses advantages, such as high tolerance to organic acid and a sufficient precursor required to synthesize 3-HP. In this study, gene manipulations, including the overexpression of genes <i>MCR-NCa</i>, <i>MCR-CCa</i>, <i>GAPNSm</i>, <i>ACC1</i> and <i>ACSSeL641P</i> and knocking out bypass genes <i>MLS1</i> and <i>CIT2</i>, leading to the glyoxylate cycle, were performed to construct a recombinant strain. Based on this, the degradation pathway of 3-HP in <i>Y. lipolytica</i> was discovered, and relevant genes <i>MMSDH</i> and <i>HPDH</i> were knocked out. To our knowledge, this study is the first to produce 3-HP in <i>Y. lipolytica</i>. The yield of 3-HP in recombinant strain Po1f-NC-14 in shake flask fermentation reached 1.128 g·L^-1, and the yield in fed-batch fermentation reached 16.23 g·L^-1. These results are highly competitive compared to other yeast chassis cells. This study creates the foundation for the production of 3-HP in <i>Y. lipolytica</i> and also provides a reference for further research in the future.