Metabolic engineering of methylotrophic Pichia pastoris for the production of β-alanine

Abstract

Abstractβ-Alanine (3-aminopropionic acid) is the only naturally occurring β-amino acid and an important precursor for the synthesis of a variety of nitrogen-containing chemicals. Fermentative production of β-alanine from renewable feedstocks such as glucose has attracted significant interest in recent years. Methanol has become an emerging and promising renewable feedstock for biomanufacturing as an alternative to glucose. In this work, we demonstrated the feasibility of β-alanine production from methanol using Pichia pastoris (Komagataella phaffii) as a methylotrophic cell factory. L-Aspartate-α-decarboxylases (ADCs) from different sources were screened and expressed in P. pastoris, followed by the optimization of aspartate decarboxylation by increasing the ADC copy number and C4 precursor supply via the overexpression of aspartate dehydrogenase. The production potential of the best strain was further evaluated in a 1-L fermenter, and a β-alanine titer of 5.6 g/L was obtained. To our best knowledge, this is the highest metabolite production titer ever reached in P. pastoris using methanol as the substrate. Graphic abstract

Methylotrophy of Pichia pastoris: Current Advances, Applications, and Future Perspectives for Methanol-Based Biomanufacturing

“…Further optimization was carried out by enhancing ADC expression strength, together with improving supply of precursor aspartate via the overexpression of aspartate dehydrogenase. The final engineered P. pastoris could produce 5.6 g/L of β-alanine from methanol . This is the first study on the synthesis of amino acids from methanol by the engineering of P. pastoris , exhibiting its great potential in amino acid production from methanol …”

“…64 β-Alanine has been widely applied as a precursor to produce a variate of nitrogen-containing chemicals for applications in food additives, pharmaceuticals, or polymer materials. 65 Its production from methanol has been achieved in engineered P. pastoris by expressing different sources of L-aspartate-α-decarboxylase (ADC). Further optimization was carried out by enhancing ADC expression strength, together with improving supply of precursor aspartate via the overexpression of aspartate dehydrogenase.…”

“…The final engineered P. pastoris could produce 5.6 g/L of β-alanine from methanol. 65 This is the first study on the synthesis of amino acids from methanol by the engineering of P. pastoris, exhibiting its great potential in amino acid production from methanol. 65 Fatty Acid and Derivatives.…”

“…65 This is the first study on the synthesis of amino acids from methanol by the engineering of P. pastoris, exhibiting its great potential in amino acid production from methanol. 65 Fatty Acid and Derivatives. Fatty acids are important platform chemicals that could serve as precursors to produce biofuels or various valuable oleochemicals.…”

“…β-Alanine has been widely applied as a precursor to produce a variate of nitrogen-containing chemicals for applications in food additives, pharmaceuticals, or polymer materials . Its production from methanol has been achieved in engineered P. pastoris by expressing different sources of l -aspartate-α-decarboxylase (ADC).…”