Establishing a synthetic pathway for high-level production of 3-hydroxypropionic acid in Saccharomyces cerevisiae via β-alanine

Abstract

Microbial fermentation of renewable feedstocks into plastic monomers can decrease our fossil dependence and reduce global CO2 emissions. 3-Hydroxypropionic acid (3HP) is a potential chemical building block for sustainable production of superabsorbent polymers and acrylic plastics. With the objective of developing Saccharomyces cerevisiae as an efficient cell factory for high-level production of 3HP, we identified the β-alanine biosynthetic route as the most economically attractive according to the metabolic modeling. We engineered and optimized a synthetic pathway for de novo biosynthesis of β-alanine and its subsequent conversion into 3HP using a novel β-alanine-pyruvate aminotransferase discovered in Bacillus cereus. The final strain produced 3HP at a titer of 13.7±0.3gL(-1) with a 0.14±0.0C-molC-mol(-1) yield on glucose in 80h in controlled fed-batch fermentation in mineral medium at pH 5, and this work therefore lays the basis for developing a process for biological 3HP production.

Advances in Metabolic Engineering of Saccharomyces cerevisiae for the Production of Industrially and Clinically Important Chemicals

“…The inal strain yielded 3-HP at a titer of 13.7 g/L in glucose-limited fed-batch cultivation. In a similar fashion, production of 3-HP via both malonyl-CoA and β-alanine pathway was reported in a xylose-utilizing S. cerevisiae [24].…”

“…Although there are biological pathways to 3-HP via either glycerol, lactate, malonyl-CoA or β-alanine intermediates, no organism is known to produce it as an end product [22]. The pathways based on malonyl-CoA and β-alanine have been constructed in S. cerevisiae [23,24]. Chen et al evaluated diferent malonyl-CoA reductases.…”

iSCHRUNK – In Silico Approach to Characterization and Reduction of Uncertainty in the Kinetic Models of Genome-scale Metabolic Networks

“…Genome--scale stoichiometric models (GEMs) of metabolism are now widely available for many organisms (Henry et al, 2010;Herrgard et al, 2008;Orth et al, 2011;Osterlund et al, 2013;Sohn et al, 2010;Thiele et al, 2013) and they have been used in studies of cellular physiology and metabolic engineering (Asadollahi et al, 2009;Borodina et al, 2015;Bro et al, 2006;Dash et al, 2014;King and Feist, 2014;Snitkin et al, 2008). However, these models are not suitable for predicting the responses of metabolism to changes in enzyme expression because they are lacking information about enzyme kinetics (Miskovic and Hatzimanikatis, 2010).…”

Identification of metabolic engineering targets for the enhancement of 1,4-butanediol production in recombinant E. coli using large-scale kinetic models

“…Microbial cell factories are becoming a norm for commercially viable production of chemicals for pharmaceutical, biotechnology, food and beverage industries (Borodina et al, 2015;Chen and Nielsen, 2013;Choi et al, 2015;Lee et al, 2012;Pfleger et al, 2015). However, engineering of microbial cell factories requires a simultaneous optimization of several criteria such as productivity, yield, titer, stress tolerance, all the while retaining the efficient, cost-effective and robust process.…”