Cargando…

Evaluation of synthetic formaldehyde and methanol assimilation pathways in Yarrowia lipolytica

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,...

Descripción completa

Detalles Bibliográficos
Autores principales: Vartiainen, Eija, Blomberg, Peter, Ilmén, Marja, Andberg, Martina, Toivari, Mervi, Penttilä, Merja
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6918578/
https://www.ncbi.nlm.nih.gov/pubmed/31890234
http://dx.doi.org/10.1186/s40694-019-0090-9
_version_ 1783480617882615808
author Vartiainen, Eija
Blomberg, Peter
Ilmén, Marja
Andberg, Martina
Toivari, Mervi
Penttilä, Merja
author_facet Vartiainen, Eija
Blomberg, Peter
Ilmén, Marja
Andberg, Martina
Toivari, Mervi
Penttilä, Merja
author_sort Vartiainen, Eija
collection PubMed
description 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 CO(2) 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.
format Online
Article
Text
id pubmed-6918578
institution National Center for Biotechnology Information
language English
publishDate 2019
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-69185782019-12-30 Evaluation of synthetic formaldehyde and methanol assimilation pathways in Yarrowia lipolytica Vartiainen, Eija Blomberg, Peter Ilmén, Marja Andberg, Martina Toivari, Mervi Penttilä, Merja Fungal Biol Biotechnol Research 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 CO(2) 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. BioMed Central 2019-12-17 /pmc/articles/PMC6918578/ /pubmed/31890234 http://dx.doi.org/10.1186/s40694-019-0090-9 Text en © The Author(s) 2019 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Vartiainen, Eija
Blomberg, Peter
Ilmén, Marja
Andberg, Martina
Toivari, Mervi
Penttilä, Merja
Evaluation of synthetic formaldehyde and methanol assimilation pathways in Yarrowia lipolytica
title Evaluation of synthetic formaldehyde and methanol assimilation pathways in Yarrowia lipolytica
title_full Evaluation of synthetic formaldehyde and methanol assimilation pathways in Yarrowia lipolytica
title_fullStr Evaluation of synthetic formaldehyde and methanol assimilation pathways in Yarrowia lipolytica
title_full_unstemmed Evaluation of synthetic formaldehyde and methanol assimilation pathways in Yarrowia lipolytica
title_short Evaluation of synthetic formaldehyde and methanol assimilation pathways in Yarrowia lipolytica
title_sort evaluation of synthetic formaldehyde and methanol assimilation pathways in yarrowia lipolytica
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6918578/
https://www.ncbi.nlm.nih.gov/pubmed/31890234
http://dx.doi.org/10.1186/s40694-019-0090-9
work_keys_str_mv AT vartiaineneija evaluationofsyntheticformaldehydeandmethanolassimilationpathwaysinyarrowialipolytica
AT blombergpeter evaluationofsyntheticformaldehydeandmethanolassimilationpathwaysinyarrowialipolytica
AT ilmenmarja evaluationofsyntheticformaldehydeandmethanolassimilationpathwaysinyarrowialipolytica
AT andbergmartina evaluationofsyntheticformaldehydeandmethanolassimilationpathwaysinyarrowialipolytica
AT toivarimervi evaluationofsyntheticformaldehydeandmethanolassimilationpathwaysinyarrowialipolytica
AT penttilamerja evaluationofsyntheticformaldehydeandmethanolassimilationpathwaysinyarrowialipolytica