Replication data for research article "Engineering the synthetic β-alanine pathway in Komagataella phaffii for conversion of methanol into 3-hydroxypropionic acid"

Supplementary files of research article "Engineering the synthetic β-alanine pathway in Komagataella phaffii for conversion of methanol into 3-hydroxypropionic acid". This research paper performs a theoretical comparison between the the two main metabolic pathways towards 3-hydroxypropionic (3-HP) acid production, and describes the establishment of the synthetic β-alanine pathway in Komagataella phaffii for conversion of methanol into 3-HP. Several strains overexpressing the β-alanine pathway were constructed and tested on 24 deep-well plate cultivations containing buffered minimal medium (BMM) with methanol.Three independent transformants from each strain were tested. Final biomass concentration as well as methanol and 3-HP quantification were determined at the end of the cultures. Series of growth kinetics experiments were performed by cultivating a representative clone from each strain in shake-flask cultures using BMM to determine the maximum specific growth rate of the different strains.

The best performing strains were further evaluated in aerobic fed-batch reactors following a pre-programmed exponential feeding strategy for controlled growth rate. Offline and online state variables during the fed-batch phase of the bioreactor-scale experiments were monitored and recorded. Nuclear Magnetic Resonance (NMR) spectroscopy was used to perform the exometabolome profiling analyses of the supernatant samples from early, mid, and late methanol-feeding phase. All experiments were carried in duplicates.

The files attached herein contain: the diagrams and stoichiometric balances of the malonyl-CoA and β-alanine pathways, the raw data obtained from the 24 deep-well plate cultivations and cell growth kinetics experiments, the NMR spectra for different strains' exometabolome, the raw and processed data obtained from the bioreactor-scale experiments, the description of the molecular cloning materials and methods, and the raw data of the online monitored standard process parameters.