Comparative lipid production by oleaginous yeasts in hydrolyzates of lignocellulosic biomass and process strategy for high titers

Patricia J. Slininger, Bruce S. Dien, Cletus P. Kurtzman, Bryan R. Moser, Erica L. Bakota, Stephanie R. Thompson, Patricia J. O'Bryan, Michael A. Cotta, Venkatesh Balan, Mingjie Jin, Leonardo da Costa Sousa, Bruce E. Dale

Research output: Contribution to journalArticle

  • 11 Citations

Abstract

Oleaginous yeasts can convert sugars to lipids with fatty acid profiles similar to those of vegetable oils, making them attractive for production of biodiesel. Lignocellulosic biomass is an attractive source of sugars for yeast lipid production because it is abundant, potentially low cost, and renewable. However, lignocellulosic hydrolyzates are laden with byproducts which inhibit microbial growth and metabolism. With the goal of identifying oleaginous yeast strains able to convert plant biomass to lipids, we screened 32 strains from the ARS Culture Collection, Peoria, IL to identify four robust strains able to produce high lipid concentrations from both acid and base-pretreated biomass. The screening was arranged in two tiers using undetoxified enzyme hydrolyzates of ammonia fiber expansion (AFEX)-pretreated cornstover as the primary screening medium and acid-pretreated switch grass as the secondary screening medium applied to strains passing the primary screen. Hydrolyzates were prepared at ∼18-20% solids loading to provide ∼110g/L sugars at ∼56:39:5 mass ratio glucose:xylose:arabinose. A two stage process boosting the molar C:N ratio from 60 to well above 400 in undetoxified switchgrass hydrolyzate was optimized with respect to nitrogen source, C:N, and carbon loading. Using this process three strains were able to consume acetic acid and nearly all available sugars to accumulate 50-65% of cell biomass as lipid (w/w), to produce 25-30g/L lipid at 0.12-0.22g/L/h and 0.13-0.15g/g or 39-45% of the theoretical yield at pH 6 and 7, a performance unprecedented in lignocellulosic hydrolyzates. Three of the top strains have not previously been reported for the bioconversion of lignocellulose to lipids. The successful identification and development of top-performing lipid-producing yeast in lignocellulose hydrolyzates is expected to advance the economic feasibility of high quality biodiesel and jet fuels from renewable biomass, expanding the market potential for lignocellulose-derived fuels beyond ethanol for automobiles to the entire U.S. transportation market.

Original languageEnglish (US)
JournalBiotechnology and Bioengineering
DOIs
StateAccepted/In press - 2016
Externally publishedYes

Profile

Biomass
Yeasts
Lipids
Panthera
Yeast
HLA Antigens
Carbohydrates
Sugars
Carbamoyl-Phosphate Synthase (Glutamine-Hydrolyzing)
Screening
Dental Materials
Biofuels
Acids
Biodiesel
Automobiles
Arabinose
Plant Oils
Xylose
Poaceae
Ammonia

Keywords

  • AFEX-pretreated corn stover
  • Biodiesel
  • Dilute acid-pretreated switchgrass
  • Renewable energy
  • Triacylglycerol

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Cite this

Comparative lipid production by oleaginous yeasts in hydrolyzates of lignocellulosic biomass and process strategy for high titers. / Slininger, Patricia J.; Dien, Bruce S.; Kurtzman, Cletus P.; Moser, Bryan R.; Bakota, Erica L.; Thompson, Stephanie R.; O'Bryan, Patricia J.; Cotta, Michael A.; Balan, Venkatesh; Jin, Mingjie; Sousa, Leonardo da Costa; Dale, Bruce E.

In: Biotechnology and Bioengineering, 2016.

Research output: Contribution to journalArticle

Slininger, PJ, Dien, BS, Kurtzman, CP, Moser, BR, Bakota, EL, Thompson, SR, O'Bryan, PJ, Cotta, MA, Balan, V, Jin, M, Sousa, LDC & Dale, BE 2016, 'Comparative lipid production by oleaginous yeasts in hydrolyzates of lignocellulosic biomass and process strategy for high titers' Biotechnology and Bioengineering. DOI: 10.1002/bit.25928

Slininger, Patricia J.; Dien, Bruce S.; Kurtzman, Cletus P.; Moser, Bryan R.; Bakota, Erica L.; Thompson, Stephanie R.; O'Bryan, Patricia J.; Cotta, Michael A.; Balan, Venkatesh; Jin, Mingjie; Sousa, Leonardo da Costa; Dale, Bruce E. / Comparative lipid production by oleaginous yeasts in hydrolyzates of lignocellulosic biomass and process strategy for high titers.

In: Biotechnology and Bioengineering, 2016.

Research output: Contribution to journalArticle

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abstract = "Oleaginous yeasts can convert sugars to lipids with fatty acid profiles similar to those of vegetable oils, making them attractive for production of biodiesel. Lignocellulosic biomass is an attractive source of sugars for yeast lipid production because it is abundant, potentially low cost, and renewable. However, lignocellulosic hydrolyzates are laden with byproducts which inhibit microbial growth and metabolism. With the goal of identifying oleaginous yeast strains able to convert plant biomass to lipids, we screened 32 strains from the ARS Culture Collection, Peoria, IL to identify four robust strains able to produce high lipid concentrations from both acid and base-pretreated biomass. The screening was arranged in two tiers using undetoxified enzyme hydrolyzates of ammonia fiber expansion (AFEX)-pretreated cornstover as the primary screening medium and acid-pretreated switch grass as the secondary screening medium applied to strains passing the primary screen. Hydrolyzates were prepared at ∼18-20% solids loading to provide ∼110g/L sugars at ∼56:39:5 mass ratio glucose:xylose:arabinose. A two stage process boosting the molar C:N ratio from 60 to well above 400 in undetoxified switchgrass hydrolyzate was optimized with respect to nitrogen source, C:N, and carbon loading. Using this process three strains were able to consume acetic acid and nearly all available sugars to accumulate 50-65% of cell biomass as lipid (w/w), to produce 25-30g/L lipid at 0.12-0.22g/L/h and 0.13-0.15g/g or 39-45% of the theoretical yield at pH 6 and 7, a performance unprecedented in lignocellulosic hydrolyzates. Three of the top strains have not previously been reported for the bioconversion of lignocellulose to lipids. The successful identification and development of top-performing lipid-producing yeast in lignocellulose hydrolyzates is expected to advance the economic feasibility of high quality biodiesel and jet fuels from renewable biomass, expanding the market potential for lignocellulose-derived fuels beyond ethanol for automobiles to the entire U.S. transportation market.",
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AU - Moser,Bryan R.

AU - Bakota,Erica L.

AU - Thompson,Stephanie R.

AU - O'Bryan,Patricia J.

AU - Cotta,Michael A.

AU - Balan,Venkatesh

AU - Jin,Mingjie

AU - Sousa,Leonardo da Costa

AU - Dale,Bruce E.

PY - 2016

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N2 - Oleaginous yeasts can convert sugars to lipids with fatty acid profiles similar to those of vegetable oils, making them attractive for production of biodiesel. Lignocellulosic biomass is an attractive source of sugars for yeast lipid production because it is abundant, potentially low cost, and renewable. However, lignocellulosic hydrolyzates are laden with byproducts which inhibit microbial growth and metabolism. With the goal of identifying oleaginous yeast strains able to convert plant biomass to lipids, we screened 32 strains from the ARS Culture Collection, Peoria, IL to identify four robust strains able to produce high lipid concentrations from both acid and base-pretreated biomass. The screening was arranged in two tiers using undetoxified enzyme hydrolyzates of ammonia fiber expansion (AFEX)-pretreated cornstover as the primary screening medium and acid-pretreated switch grass as the secondary screening medium applied to strains passing the primary screen. Hydrolyzates were prepared at ∼18-20% solids loading to provide ∼110g/L sugars at ∼56:39:5 mass ratio glucose:xylose:arabinose. A two stage process boosting the molar C:N ratio from 60 to well above 400 in undetoxified switchgrass hydrolyzate was optimized with respect to nitrogen source, C:N, and carbon loading. Using this process three strains were able to consume acetic acid and nearly all available sugars to accumulate 50-65% of cell biomass as lipid (w/w), to produce 25-30g/L lipid at 0.12-0.22g/L/h and 0.13-0.15g/g or 39-45% of the theoretical yield at pH 6 and 7, a performance unprecedented in lignocellulosic hydrolyzates. Three of the top strains have not previously been reported for the bioconversion of lignocellulose to lipids. The successful identification and development of top-performing lipid-producing yeast in lignocellulose hydrolyzates is expected to advance the economic feasibility of high quality biodiesel and jet fuels from renewable biomass, expanding the market potential for lignocellulose-derived fuels beyond ethanol for automobiles to the entire U.S. transportation market.

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