Comparative Lipid Production by Oleaginous Yeasts in Hydrolyzates of Lignocellulosic Biomass and Process Strategy for High Titers

Authors

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

Document Type

Article

Date of this Version

2016

Citation

Biotechnol. Bioeng. 2016;9999: pp. 1–15. DOI 10.1002/bit.25928

Comments

U. S. government work.

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 approximately 18–20% solids loading to provide approximately 110 g/L sugars at approximately 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 25–30 g/L lipid at 0.12– 0.22 g/L/h and 0.13–0.15 g/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 lipidproducing 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.