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

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