Comparative transcriptome and metabolome analyses reveal the methanol dissimilation pathway of Pichia pastoris

Yu, Yifan; Yang, Jingli; Zhao, Fang; Lin, Ying; Han, Sang‐Ik

doi:10.1186/s12864-022-08592-8

Cited by 22 publications

(16 citation statements)

References 61 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In the later stage of fermentation, methanol was consumed as the carbon source. According to the metabolism way of methanol in the P. pastoris yeast, after entering the cell, it will be first oxidized to formaldehyde by alcohol oxidase, and then formaldehyde will be oxidized by formaldehyde dehydrogenase, S-hydroxymethyl glutathione hydrolase, and formate dehydrogenase in the dissimilation pathway to generate CO 2 . However, carbon balance analysis shows that the CO 2 emissions is very low when the strain is active in the early stages .…”

Section: Results and Discussionmentioning

confidence: 99%

“…According to the metabolism way of methanol in the P. pastoris yeast, after entering the cell, it will be first oxidized to formaldehyde by alcohol oxidase, and then formaldehyde will be oxidized by formaldehyde dehydrogenase, S-hydroxymethyl glutathione hydrolase, and formate dehydrogenase in the dissimilation pathway to generate CO 2 . 62 However, carbon balance analysis shows that the CO 2 emissions is very low when the strain is active in the early stages. 63 More CO 2 will be produced only when methanol is excessively supplied, and the strain performance declines in the later stages of fermentation.…”

Section: Effect Of Abts Concentration On Cell Performance For Electri...mentioning

confidence: 99%

See 1 more Smart Citation

Laccase-Mediated Oxygen Reduction in Liquid Flow Fuel Cells for Efficient Oxidation of Biomass-Derived Aldehydes with Co-Generation of Electricity

Liu,

Dai,

Ouyang

et al. 2024

ACS Sustainable Chem. Eng.

View full text Add to dashboard Cite

Laccase was produced by heterologous expression of gene lacc6 from Pleurotus otreatus HAUCC 162 and then employed as an efficient catalyst to increase the kinetics of the oxygen reduction reaction in a liquid flow fuel cell (LFFC). An electron transport chain (ETC) was constructed in the LFFC with Ag 2 O as an anode electrocatalyst and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and laccase as the cathode electron mediators. The LFFC could well convert furfural to furoic acid with co-generation of electricity. Under the optimized cathodic conditions of 40 °C, pH 4.0, 50 mM ABTS, and laccase loading of 300 U/mmol ABTS, a P max of 70.8 mW/cm 2 could be obtained. For furoic acid production, a 91% yield could be obtained under the anodic condition of 100 mM furfural and 1.5 M KOH. The electrons released from the oxidation of furfural on the Ag 2 O catalyst could be well transferred to that external circuit with a Faraday efficiency of 96.2%. This work thus can provide a novel route for clean production of biomass-based chemicals and electricity by a combination of chemical, electrochemical, and biological processes.

show abstract

Section: Results and Discussionmentioning

confidence: 99%

Section: Effect Of Abts Concentration On Cell Performance For Electri...mentioning

confidence: 99%

Laccase-Mediated Oxygen Reduction in Liquid Flow Fuel Cells for Efficient Oxidation of Biomass-Derived Aldehydes with Co-Generation of Electricity

Liu,

Dai,

Ouyang

et al. 2024

ACS Sustainable Chem. Eng.

View full text Add to dashboard Cite

show abstract

“…Methanol dissimilation pathway consumes formaldehyde toward CO 2 production with NADH regeneration, which might lead to the low efficiency of methanol bioconversion. , We thus endeavored to downregulate the methanol dissimilation pathway. It is impossible to completely block the methanol dissimilation pathway by deleting the key genes FLD1 encoding formaldehyde dehydrogenase or FDH1 encoding formate dehydrogenase (Figure A), which is essential for providing NADH to support cell growth of P.…”

Section: Resultsmentioning

confidence: 99%

Efficient Bioproduction of 3-Hydroxypropionic Acid from Methanol by a Synthetic Yeast Cell Factory

Cai

Gao

et al. 2023

ACS Sustainable Chem. Eng.

View full text Add to dashboard Cite

Methanol is an ideal feedstock for bio-manufacturing chemicals without the dependence of sugars and the competition of arable lands. We here engineered an industrial yeast Pichia pastoris to efficiently produce 3-hydroxypropionic acid (3-HP) from sole methanol as a carbon source by using a malonyl-CoA-derived pathway. Optimizing the expression of malonyl-CoA reductase gene MCR from Chloroflexus aurantiacus and enhancing the supply of precursors and NADPH enabled 3-HP production of 1.5 g/L. To avoid the time-consuming genetic manipulation, metabolically transforming a free fatty acid (FFA)-overproducing strain toward 3-HP biosynthesis results in a 3-HP production of 1.9 g/L in a shake flask. Through further downregulation of methanol dissimilation, 3-HP production was improved to 2.2 g/L. Subsequent fed-batch cultivation in bioreactors achieved a remarkable 3-HP production of 48.2 g/L from minimal medium with a yield of 0.23 g/g methanol. Notably, this represents the highest reported 3-HP production from one-carbon (C1) feedstocks and is comparable to that from sugar in yeast. The high-level 3-HP production from methanol highlights the potential of P. pastoris as a workhorse for methanol biotransformation. Furthermore, the strategies presented in this study could be applied for production of other acetyl-CoA derivatives from methanol in P. pastoris.

show abstract

“…The natural methanol metabolism in P. pastoris involves carbon dioxide loss via the formaldehyde dissimilatory pathway through glutathione-dependent formaldehyde dehydrogenase (FLD), S-formyl glutathione hydrolase (FGH), and formate dehydrogenase (FDH), the main source of cellular redox power for support of cell growth. The inactivation of formaldehyde dissimilation for carbon conservation through deletion of FLD or FDH would signi cantly reduce the biomass yield of P. pastoris on methanol 30 . The transfer of the entire methanol utilization pathway in P. pastoris into other model yeasts including S. cerevisiae and Yarrowia lipolytica has been demonstrated to be a di cult and inconvenient strategy in developing full synthetic methylotrophic yeast.…”

Section: Resultsmentioning

confidence: 99%

Engineering Yeasts to Grow Solely on Methanol or Formic acid coupled with CO2 fixation

Guo

Zhang

Wang

et al. 2023

Preprint

View full text Add to dashboard Cite

Synthetic microorganisms capable of using one-carbon compounds, such as methanol, formic acid or carbon dioxide, are of increasing interest. In this study, we converted the yeasts of Pichia pastoris and Saccharomyces cerevisiae to both synthetic methylotroph and formatotroph, allowing them to grow on methanol and formic acid alone coupled with CO2 fixation through a synthetic C1-compound assimilation pathway (MFORG pathway). This pathway consists of a methanol-formic acid oxidation module and the reductive glycine pathway. We first assembled the MFORG pathway in P. pastoris using only native enzymes, followed by overexpression of genes in the reductive glycine pathway, blocking the native methanol assimilation pathway, and compartmentalizing the methanol oxidation module. These modifications successfully redesigned the native methylotrophic yeast P. pastoris to grow on both methanol and formic acid, where higher growth rate and yield on methanol was obtained compared to the wild-type strain. We then introduced the MFORG pathway from P. pastoris into the model yeast S. cerevisiae, establishing full synthetic methylotrophy and formatotrophy in this organism. The resulting strain was able to successfully grow on methanol or formic acid alone with consumption rates of 24 mg/L*h and 15.2 mg/L*h, respectively. The CO2 fixation ability of synthetic P. pastoris and S. cerevisiae through the MFORG pathway was confirmed by 13C-tracer analysis. Finally, production of 5-aminolevulinic acid and lactic acid with methanol as the sole carbon source was demonstrated using synthetic P. pastoris and S. cerevisiae, indicating the potential of yeasts as promising hosts for biochemical production from various one-carbon compounds.

show abstract

Comparative transcriptome and metabolome analyses reveal the methanol dissimilation pathway of Pichia pastoris

Cited by 22 publications

References 61 publications

Laccase-Mediated Oxygen Reduction in Liquid Flow Fuel Cells for Efficient Oxidation of Biomass-Derived Aldehydes with Co-Generation of Electricity

Laccase-Mediated Oxygen Reduction in Liquid Flow Fuel Cells for Efficient Oxidation of Biomass-Derived Aldehydes with Co-Generation of Electricity

Efficient Bioproduction of 3-Hydroxypropionic Acid from Methanol by a Synthetic Yeast Cell Factory

Engineering Yeasts to Grow Solely on Methanol or Formic acid coupled with CO2 fixation

Contact Info

Product

Resources

About