Evaluation of synthetic formaldehyde and methanol assimilation pathways in Yarrowia lipolytica

Eija Vartiainen, Peter Blomberg, Marja Ilmén, Martina Andberg, Mervi Toivari (Corresponding Author), Merja Penttilä

Research output: Contribution to journalArticleScientificpeer-review

Abstract

Background: Crude glycerol coming from biodiesel production is an attractive carbon source for biological production of chemicals. The major impurity in preparations of crude glycerol is methanol, which is toxic for most microbes. Development of microbes, which would not only tolerate the methanol, but also use it as co-substrate, would increase the feasibility of bioprocesses using crude glycerol as substrate.
Results: To prevent methanol conversion to CO2 via formaldehyde and formate, the formaldehyde dehydrogenase (FLD) gene was identified in and deleted from Yarrowia lipolytica. The deletion strain was able to convert methanol to formaldehyde without expression of heterologous methanol dehydrogenases. Further, it was shown that expression of heterologous formaldehyde assimilating enzymes could complement the deletion of FLD. The expression of either 3-hexulose-6-phosphate synthase (HPS) enzyme of ribulose monosphosphate pathway or dihydroxyacetone synthase (DHAS) enzyme of xylulose monosphosphate pathway restored the formaldehyde tolerance of the formaldehyde sensitive Δfld1 strain.
Conclusions: In silico, the expression of heterologous formaldehyde assimilation pathways enable Y. lipolytica to use methanol as substrate for growth and metabolite production. In vivo, methanol was shown to be converted to formaldehyde and the enzymes of formaldehyde assimilation were actively expressed in this yeast. However, further development is required to enable Y. lipolytica to efficiently use methanol as co-substrate with glycerol.
Original languageEnglish
Article number27
JournalFungal Biology and Biotechnology
Volume6
DOIs
Publication statusPublished - 17 Dec 2019
MoE publication typeA1 Journal article-refereed

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Yarrowia
Formaldehyde
Methanol
glutathione-independent formaldehyde dehydrogenase
Glycerol
formaldehyde transketolase
Enzymes
Formate Dehydrogenases
Xylulose
Biofuels
Poisons
Computer Simulation
Carbon
Yeasts

Keywords

  • Yarrowia lipolytica
  • Crude glycerol,
  • Methano
  • formaldehyde exposure

Cite this

@article{ab7723070d8b49abac1c2289d2238cf3,
title = "Evaluation of synthetic formaldehyde and methanol assimilation pathways in Yarrowia lipolytica",
abstract = "Background: Crude glycerol coming from biodiesel production is an attractive carbon source for biological production of chemicals. The major impurity in preparations of crude glycerol is methanol, which is toxic for most microbes. Development of microbes, which would not only tolerate the methanol, but also use it as co-substrate, would increase the feasibility of bioprocesses using crude glycerol as substrate.Results: To prevent methanol conversion to CO2 via formaldehyde and formate, the formaldehyde dehydrogenase (FLD) gene was identified in and deleted from Yarrowia lipolytica. The deletion strain was able to convert methanol to formaldehyde without expression of heterologous methanol dehydrogenases. Further, it was shown that expression of heterologous formaldehyde assimilating enzymes could complement the deletion of FLD. The expression of either 3-hexulose-6-phosphate synthase (HPS) enzyme of ribulose monosphosphate pathway or dihydroxyacetone synthase (DHAS) enzyme of xylulose monosphosphate pathway restored the formaldehyde tolerance of the formaldehyde sensitive Δfld1 strain.Conclusions: In silico, the expression of heterologous formaldehyde assimilation pathways enable Y. lipolytica to use methanol as substrate for growth and metabolite production. In vivo, methanol was shown to be converted to formaldehyde and the enzymes of formaldehyde assimilation were actively expressed in this yeast. However, further development is required to enable Y. lipolytica to efficiently use methanol as co-substrate with glycerol.",
keywords = "Yarrowia lipolytica, Crude glycerol,, Methano, formaldehyde exposure",
author = "Eija Vartiainen and Peter Blomberg and Marja Ilm{\'e}n and Martina Andberg and Mervi Toivari and Merja Penttil{\"a}",
year = "2019",
month = "12",
day = "17",
doi = "10.1186/s40694-019-0090-9",
language = "English",
volume = "6",
journal = "Fungal Biology and Biotechnology",
issn = "2054-3085",

}

TY - JOUR

T1 - Evaluation of synthetic formaldehyde and methanol assimilation pathways in Yarrowia lipolytica

AU - Vartiainen, Eija

AU - Blomberg, Peter

AU - Ilmén, Marja

AU - Andberg, Martina

AU - Toivari, Mervi

AU - Penttilä, Merja

PY - 2019/12/17

Y1 - 2019/12/17

N2 - Background: Crude glycerol coming from biodiesel production is an attractive carbon source for biological production of chemicals. The major impurity in preparations of crude glycerol is methanol, which is toxic for most microbes. Development of microbes, which would not only tolerate the methanol, but also use it as co-substrate, would increase the feasibility of bioprocesses using crude glycerol as substrate.Results: To prevent methanol conversion to CO2 via formaldehyde and formate, the formaldehyde dehydrogenase (FLD) gene was identified in and deleted from Yarrowia lipolytica. The deletion strain was able to convert methanol to formaldehyde without expression of heterologous methanol dehydrogenases. Further, it was shown that expression of heterologous formaldehyde assimilating enzymes could complement the deletion of FLD. The expression of either 3-hexulose-6-phosphate synthase (HPS) enzyme of ribulose monosphosphate pathway or dihydroxyacetone synthase (DHAS) enzyme of xylulose monosphosphate pathway restored the formaldehyde tolerance of the formaldehyde sensitive Δfld1 strain.Conclusions: In silico, the expression of heterologous formaldehyde assimilation pathways enable Y. lipolytica to use methanol as substrate for growth and metabolite production. In vivo, methanol was shown to be converted to formaldehyde and the enzymes of formaldehyde assimilation were actively expressed in this yeast. However, further development is required to enable Y. lipolytica to efficiently use methanol as co-substrate with glycerol.

AB - Background: Crude glycerol coming from biodiesel production is an attractive carbon source for biological production of chemicals. The major impurity in preparations of crude glycerol is methanol, which is toxic for most microbes. Development of microbes, which would not only tolerate the methanol, but also use it as co-substrate, would increase the feasibility of bioprocesses using crude glycerol as substrate.Results: To prevent methanol conversion to CO2 via formaldehyde and formate, the formaldehyde dehydrogenase (FLD) gene was identified in and deleted from Yarrowia lipolytica. The deletion strain was able to convert methanol to formaldehyde without expression of heterologous methanol dehydrogenases. Further, it was shown that expression of heterologous formaldehyde assimilating enzymes could complement the deletion of FLD. The expression of either 3-hexulose-6-phosphate synthase (HPS) enzyme of ribulose monosphosphate pathway or dihydroxyacetone synthase (DHAS) enzyme of xylulose monosphosphate pathway restored the formaldehyde tolerance of the formaldehyde sensitive Δfld1 strain.Conclusions: In silico, the expression of heterologous formaldehyde assimilation pathways enable Y. lipolytica to use methanol as substrate for growth and metabolite production. In vivo, methanol was shown to be converted to formaldehyde and the enzymes of formaldehyde assimilation were actively expressed in this yeast. However, further development is required to enable Y. lipolytica to efficiently use methanol as co-substrate with glycerol.

KW - Yarrowia lipolytica

KW - Crude glycerol,

KW - Methano

KW - formaldehyde exposure

U2 - 10.1186/s40694-019-0090-9

DO - 10.1186/s40694-019-0090-9

M3 - Article

VL - 6

JO - Fungal Biology and Biotechnology

JF - Fungal Biology and Biotechnology

SN - 2054-3085

M1 - 27

ER -