Metabolic engineering of Pichia pastoris for production of isopentanol (3-Methyl-1-butanol)

Siripong, Wiparat; Angela, Clara; Tanapongpipat, Sutipa; Runguphan, Weerawat

doi:10.1016/j.enzmictec.2020.109557

Cited by 31 publications

(19 citation statements)

References 39 publications

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“…Notably, the isobutanol titers produced by our engineered strains PPY0311 and PPY0312 using sugarcane trash hydrolysates remain well below the grams/liter level obtained when using pure glucose as the carbon source described in our earlier work [ 14 , 15 ]. While product titers can typically be improved by increasing the amount of carbon source in the fermentation medium (though only to a certain point and product yield might be adversely affected) [ 32 ], in our case, however, increasing the concentration of sugarcane trash hydrolysates also means letting in higher concentrations of the toxic pretreatment inhibitors which would be detrimental to yeast growth.…”

Section: Resultsmentioning

confidence: 58%

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Engineered Production of Isobutanol from Sugarcane Trash Hydrolysates in Pichia pastoris

Bumrungtham

Promdonkoy

Prabmark

et al. 2022

JoF

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Concerns over climate change have led to increased interest in renewable fuels in recent years. Microbial production of advanced fuels from renewable and readily available carbon sources has emerged as an attractive alternative to the traditional production of transportation fuels. Here, we engineered the yeast Pichia pastoris, an industrial powerhouse in heterologous enzyme production, to produce the advanced biofuel isobutanol from sugarcane trash hydrolysates. Our strategy involved overexpressing a heterologous xylose isomerase and the endogenous xylulokinase to enable the yeast to consume both C5 and C6 sugars in biomass. To enable the yeast to produce isobutanol, we then overexpressed the endogenous amino acid biosynthetic pathway and the 2-keto acid degradation pathway. The engineered strains produced isobutanol at a titer of up to 48.2 ± 1.7 mg/L directly from a minimal medium containing sugarcane trash hydrolysates as the sole carbon source. To our knowledge, this is the first demonstration of advanced biofuel production using agricultural waste-derived hydrolysates in the yeast P. pastoris. We envision that our work will pave the way for a scalable route to this advanced biofuel and further establish P. pastoris as a versatile production platform for fuels and high-value chemicals.

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Section: Resultsmentioning

confidence: 58%

“…Primers used for plasmid construction are listed in Supplementary Materials . Yeast transformation was performed as previously described using an electroporator with the following parameters: 1.5 kV, 25 μF, 200 Ω [ 15 ]. The integrative plasmids were linearized with the restriction enzyme AvrII before electroporation.…”

Section: Methodsmentioning

confidence: 99%

Engineered Production of Isobutanol from Sugarcane Trash Hydrolysates in Pichia pastoris

Bumrungtham

Promdonkoy

Prabmark

et al. 2022

JoF

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“…The levels of 2-amino-3-methyl-1-butanol, N-acetyl- l -glutamine, N-acryloylglycine, phenyl propionyl glycine, and phenethyl acetate in PA treatment were significantly lower compared with those in the CON group (Figure A 5–9 ). According to the accumulation of isobutanol concerned with converting valine by the keto-acid degradation pathway, the higher content of 2-amino-3-methyl-1-butanol in the CON group demonstrated more serious amino acid degradation compared to that in PA treatment. In addition, the study found that amino acids were easy to form acyl-CoA thioester, followed by conjugation with glycine in the presence of microorganisms .…”

Section: Results and Discussionmentioning

confidence: 99%

Relationship between Micromolecules and Quality Changes of Tilapia Fillets after Partial Freezing Treatment with Polyphenols

Wei

Zhu

Cao

et al. 2021

J. Agric. Food Chem.

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The study investigated the main characteristic micromolecular changes in tilapia fillets after partial freezing treatment with polyphenols by ultrahigh-performance liquid chromatography−quadrupole time-of-flight mass spectrometry (UHPLC−Q-TOF-MS) analysis. A total of 2121 metabolite ion features were identified. The result suggested that procyanidin treatment increased the sweet, salty, and thick peptides' contents and suppressed the formation of bitter peptides. The levels of cis-4octenedioic acid, 2-amino-heptanoic acid, indoleacrylic acid, and 2-amino-3-methyl-1-butanol in polyphenol treatments were significantly lower compared to those in the control group (P < 0.05), which delayed the formation of micromolecule of acids and alcohols associated with spoilage and inhibited the protein and lipid oxidation and degradation. Polyphenol treatments suppressed the formation of biogenic amines (lower levels of spermidine and 1-naphthylacetylspermine) and reduced fillet quality deterioration. It provided critical novel insights into the understanding of the molecular mechanism for inhibiting the quality deterioration of fillets treated with polyphenols during storage.

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“…Reports on metabolite production in fermentations of engineered P. pastoris mainly remain at the proof-of-concept stage, with titers usually lower than 1 g/L, especially when methanol is used as substrate (Gao et al 2021). Several recent studies reported gram per liter metabolites production in shake asks, with examples including isobutanol (Siripong et al 2020;Siripong et al 2018), malic acid (Guo et al 2021), and D-lactic acid (Yamada et al 2019). Unfortunately, the chemicals production potential of these recombinant P. pastoris strains was not fully evaluated by cultivation on the fermenter scale.…”

Section: Discussionmentioning

confidence: 99%

Metabolic Engineering of the Methylotrophic Yeast Pichia Pastoris (Komagataella phaffii) for the Production of Β-alanine From Methanol

Miao

Zhu

2021

Preprint

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β-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. Aspartate 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-liter fermenter, and a β-alanine titer of 5.6 g/L was obtained. To our best knowledge, this is the highest chemical production titer ever reached in P. pastoris using methanol as the substrate.

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Metabolic engineering of Pichia pastoris for production of isopentanol (3-Methyl-1-butanol)

Cited by 31 publications

References 39 publications

Engineered Production of Isobutanol from Sugarcane Trash Hydrolysates in Pichia pastoris

Engineered Production of Isobutanol from Sugarcane Trash Hydrolysates in Pichia pastoris

Relationship between Micromolecules and Quality Changes of Tilapia Fillets after Partial Freezing Treatment with Polyphenols

Metabolic Engineering of the Methylotrophic Yeast Pichia Pastoris (Komagataella phaffii) for the Production of Β-alanine From Methanol

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