Metabolic engineering of yeasts for production of bulk fermentation products from xylose

ethanol and lactic acid

C. Miller, V. Rajgarhia, Marja Ilmen, Kari Koivuranta, Laura Ruohonen, Aristos Aristidou, Merja Penttilä, Pirkko Suominen

Research output: Chapter in Book/Report/Conference proceedingConference abstract in proceedingsScientific

Abstract

Kluyveromyces marxianus, a yeast naturally assimilating but not fermenting xylose, was genetically engineered to produce ethanol from xylose efficiently. Genetic engineering included replacing the natural xylose utilization pathway via xylose reductase and xylitol dehydrogenase to xylulose by a fungal xylose isomerase converting xylose directly to xylulose. Furthermore xylulokinase was overexpressed to improve efficiency of xylose to ethanol fermentation. The resulting strain produced 37 g/L ethanol with a yield of 0.4 g/g xylose used and an ethanol production rate of 0.94 g/L*h in shake flask fermentation tests. The original strain produced xylitol very efficiently in these conditions but did not produce any ethanol. The above strain was further engineered to produce lactic acid from xylose by expressing the L-lactate dehydrogenase (LDH) from Lactobacillus helveticus. The resulting strain produced lactate from xylose in shake flask test conditions with 77% yield and 1 g/L*h productivity. Lactobacillus helveticus LDH was also introduced to Pichia stipitis, a yeast naturally fermenting xylose. Lactate was the main product from xylose in shake flask test and was produced with about 60% yield and 0.4 g/L*h productivity.
Original languageEnglish
Title of host publicationInternational Specialised Symposium on Yeasts ISSY25
Subtitle of host publicationSystems Biology of Yeasts - from Models to Applications
PublisherVTT Technical Research Centre of Finland
Pages45
ISBN (Electronic) 951-38-6308-5
ISBN (Print)951-38-6307-7
Publication statusPublished - 2006
EventInternational Specialised Symposium on Yeasts, ISSY 25 - Espoo, Finland
Duration: 18 Jun 200621 Jun 2006

Publication series

NameVTT Symposium
PublisherVTT
Number242
ISSN (Print)0357-9387
ISSN (Electronic)1455-0873

Conference

ConferenceInternational Specialised Symposium on Yeasts, ISSY 25
Abbreviated titleISSY 25
CountryFinland
CityEspoo
Period18/06/0621/06/06

Fingerprint

metabolic engineering
xylose
lactic acid
ethanol
fermentation
yeasts
xylulose
Lactobacillus helveticus
xylitol
lactate dehydrogenase
lactates
liquid state fermentation
xylose isomerase
Scheffersomyces stipitis
Kluyveromyces marxianus
ethanol fermentation
testing
ethanol production
genetic engineering

Cite this

Miller, C., Rajgarhia, V., Ilmen, M., Koivuranta, K., Ruohonen, L., Aristidou, A., ... Suominen, P. (2006). Metabolic engineering of yeasts for production of bulk fermentation products from xylose: ethanol and lactic acid. In International Specialised Symposium on Yeasts ISSY25: Systems Biology of Yeasts - from Models to Applications (pp. 45). [T30] VTT Technical Research Centre of Finland. VTT Symposium, No. 242
Miller, C. ; Rajgarhia, V. ; Ilmen, Marja ; Koivuranta, Kari ; Ruohonen, Laura ; Aristidou, Aristos ; Penttilä, Merja ; Suominen, Pirkko. / Metabolic engineering of yeasts for production of bulk fermentation products from xylose : ethanol and lactic acid. International Specialised Symposium on Yeasts ISSY25: Systems Biology of Yeasts - from Models to Applications. VTT Technical Research Centre of Finland, 2006. pp. 45 (VTT Symposium; No. 242).
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abstract = "Kluyveromyces marxianus, a yeast naturally assimilating but not fermenting xylose, was genetically engineered to produce ethanol from xylose efficiently. Genetic engineering included replacing the natural xylose utilization pathway via xylose reductase and xylitol dehydrogenase to xylulose by a fungal xylose isomerase converting xylose directly to xylulose. Furthermore xylulokinase was overexpressed to improve efficiency of xylose to ethanol fermentation. The resulting strain produced 37 g/L ethanol with a yield of 0.4 g/g xylose used and an ethanol production rate of 0.94 g/L*h in shake flask fermentation tests. The original strain produced xylitol very efficiently in these conditions but did not produce any ethanol. The above strain was further engineered to produce lactic acid from xylose by expressing the L-lactate dehydrogenase (LDH) from Lactobacillus helveticus. The resulting strain produced lactate from xylose in shake flask test conditions with 77{\%} yield and 1 g/L*h productivity. Lactobacillus helveticus LDH was also introduced to Pichia stipitis, a yeast naturally fermenting xylose. Lactate was the main product from xylose in shake flask test and was produced with about 60{\%} yield and 0.4 g/L*h productivity.",
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Miller, C, Rajgarhia, V, Ilmen, M, Koivuranta, K, Ruohonen, L, Aristidou, A, Penttilä, M & Suominen, P 2006, Metabolic engineering of yeasts for production of bulk fermentation products from xylose: ethanol and lactic acid. in International Specialised Symposium on Yeasts ISSY25: Systems Biology of Yeasts - from Models to Applications., T30, VTT Technical Research Centre of Finland, VTT Symposium, no. 242, pp. 45, International Specialised Symposium on Yeasts, ISSY 25 , Espoo, Finland, 18/06/06.

Metabolic engineering of yeasts for production of bulk fermentation products from xylose : ethanol and lactic acid. / Miller, C.; Rajgarhia, V.; Ilmen, Marja; Koivuranta, Kari; Ruohonen, Laura; Aristidou, Aristos; Penttilä, Merja; Suominen, Pirkko.

International Specialised Symposium on Yeasts ISSY25: Systems Biology of Yeasts - from Models to Applications. VTT Technical Research Centre of Finland, 2006. p. 45 T30 (VTT Symposium; No. 242).

Research output: Chapter in Book/Report/Conference proceedingConference abstract in proceedingsScientific

TY - CHAP

T1 - Metabolic engineering of yeasts for production of bulk fermentation products from xylose

T2 - ethanol and lactic acid

AU - Miller, C.

AU - Rajgarhia, V.

AU - Ilmen, Marja

AU - Koivuranta, Kari

AU - Ruohonen, Laura

AU - Aristidou, Aristos

AU - Penttilä, Merja

AU - Suominen, Pirkko

PY - 2006

Y1 - 2006

N2 - Kluyveromyces marxianus, a yeast naturally assimilating but not fermenting xylose, was genetically engineered to produce ethanol from xylose efficiently. Genetic engineering included replacing the natural xylose utilization pathway via xylose reductase and xylitol dehydrogenase to xylulose by a fungal xylose isomerase converting xylose directly to xylulose. Furthermore xylulokinase was overexpressed to improve efficiency of xylose to ethanol fermentation. The resulting strain produced 37 g/L ethanol with a yield of 0.4 g/g xylose used and an ethanol production rate of 0.94 g/L*h in shake flask fermentation tests. The original strain produced xylitol very efficiently in these conditions but did not produce any ethanol. The above strain was further engineered to produce lactic acid from xylose by expressing the L-lactate dehydrogenase (LDH) from Lactobacillus helveticus. The resulting strain produced lactate from xylose in shake flask test conditions with 77% yield and 1 g/L*h productivity. Lactobacillus helveticus LDH was also introduced to Pichia stipitis, a yeast naturally fermenting xylose. Lactate was the main product from xylose in shake flask test and was produced with about 60% yield and 0.4 g/L*h productivity.

AB - Kluyveromyces marxianus, a yeast naturally assimilating but not fermenting xylose, was genetically engineered to produce ethanol from xylose efficiently. Genetic engineering included replacing the natural xylose utilization pathway via xylose reductase and xylitol dehydrogenase to xylulose by a fungal xylose isomerase converting xylose directly to xylulose. Furthermore xylulokinase was overexpressed to improve efficiency of xylose to ethanol fermentation. The resulting strain produced 37 g/L ethanol with a yield of 0.4 g/g xylose used and an ethanol production rate of 0.94 g/L*h in shake flask fermentation tests. The original strain produced xylitol very efficiently in these conditions but did not produce any ethanol. The above strain was further engineered to produce lactic acid from xylose by expressing the L-lactate dehydrogenase (LDH) from Lactobacillus helveticus. The resulting strain produced lactate from xylose in shake flask test conditions with 77% yield and 1 g/L*h productivity. Lactobacillus helveticus LDH was also introduced to Pichia stipitis, a yeast naturally fermenting xylose. Lactate was the main product from xylose in shake flask test and was produced with about 60% yield and 0.4 g/L*h productivity.

M3 - Conference abstract in proceedings

SN - 951-38-6307-7

T3 - VTT Symposium

SP - 45

BT - International Specialised Symposium on Yeasts ISSY25

PB - VTT Technical Research Centre of Finland

ER -

Miller C, Rajgarhia V, Ilmen M, Koivuranta K, Ruohonen L, Aristidou A et al. Metabolic engineering of yeasts for production of bulk fermentation products from xylose: ethanol and lactic acid. In International Specialised Symposium on Yeasts ISSY25: Systems Biology of Yeasts - from Models to Applications. VTT Technical Research Centre of Finland. 2006. p. 45. T30. (VTT Symposium; No. 242).