Irradiation of Yarrowia lipolytica NRRL YB-567 creating novel strains with enhanced ammonia and oil production on protein and carbohydrate substrates
Irradiation of Yarrowia lipolytica NRRL YB-567 creating novel strains with enhanced ammonia and oil production on protein and carbohydrate substrates
Mitch R. Lindquist 0 1 2 3 4 5 6
Juan Carlos López-Núñez 0 1 2 3 4 5 6
Marjorie A. Jones 0 1 2 3 4 5 6
Elby J. Cox 0 1 2 3 4 5 6
Rebecca J. Pinkelman 0 1 2 3 4 5 6
Sookie S. Bang 0 1 2 3 4 5 6
Bryan R. Moser 0 1 2 3 4 5 6
Michael A. Jackson 0 1 2 3 4 5 6
Loren B. Iten 0 1 2 3 4 5 6
Cletus P. Kurtzman 0 1 2 3 4 5 6
Kenneth M. Bischoff 0 1 2 3 4 5 6
Siqing Liu 0 1 2 3 4 5 6
Nasib Qureshi 0 1 2 3 4 5 6
Kenneth Tasaki 0 1 2 3 4 5 6
Joseph O. Rich 0 1 2 3 4 5 6
Michael A. Cotta 0 1 2 3 4 5 6
Badal C. Saha 0 1 2 3 4 5 6
Stephen R. Hughes 0 1 2 3 4 5 6
0 South Dakota School of Mines & Technology, Chemical and Biological Engineering , 501 East Saint Joseph Street, Rapid City, SD 57701-3995 , USA
1 National Coffee Research Centre - Cenicafe, National Federation of Coffee Growers of Colombia - FNC , Cenicafé Planalto Km 4 vía Antigua Chinchiná, Manizales, Caldas , Colombia
2 United States Department of Agriculture (USDA), Agricultural Research Service (ARS), National Center for Agricultural Utilization Research (NCAUR), Renewable Product Technology Research Unit , 1815 North University Street, Peoria, IL 61604 , USA
3 Mitsubishi Chemical, USMC Research & Innovation , 410 Palos Verdes Blvd, Redondo Beach, CA 90277 , USA
4 USDA, ARS, NCAUR, Bacterial Foodborne Pathogens and Mycology Research Unit , 1815 North University Street, Peoria, IL 61604 , USA
5 USDA, ARS, NCAUR, Bioenergy Research Unit , 1815 North University Street, Peoria, IL 61604 , USA
6 USDA, ARS, NCAUR, Bio-oils Research Unit , 1815 North University Street, Peoria, IL 61604 , USA
Increased interest in sustainable production of renewable diesel and other valuable bioproducts is redoubling efforts to improve economic feasibility of microbial-based oil production. Yarrowia lipolytica is capable of employing a wide variety of substrates to produce oil and valuable co-products. We irradiated Y. lipolytica NRRL YB-567 with UV-C to enhance ammonia (for fertilizer) and lipid (for biodiesel) production on low-cost protein and carbohydrate substrates. The resulting strains were screened for ammonia and oil production using color intensity of indicators on plate assays. Seven mutant strains were selected (based on ammonia assay) and further evaluated for growth rate, ammonia and oil production, soluble protein content, and morphology when grown on liver infusion medium (without sugars), and for growth on various Mention of trade names or commercial products in this article is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the US Department of Agriculture. USDA is an equal opportunity provider and employer.
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* Stephen R. Hughes
substrates. Strains were identified among these mutants
that had a faster doubling time, produced higher maximum
ammonia levels (enzyme assay) and more oil (Sudan Black
assay), and had higher maximum soluble protein levels
(Bradford assay) than wild type. When grown on plates
with substrates of interest, all mutant strains showed
similar results aerobically to wild-type strain. The mutant
strain with the highest oil production and the fastest
doubling time was evaluated on coffee waste medium. On this
medium, the strain produced 0.12 g/L ammonia and 0.20 g/
L 2-phenylethanol, a valuable fragrance/flavoring, in
addition to acylglycerols (oil) containing predominantly C16
and C18 residues. These mutant strains will be investigated
further for potential application in commercial biodiesel
production.
One of the major challenges facing commercial production of
biofuels and bioproducts is cost-effective utilization,
detoxification, and processing of biomass and other inexpensive carbon
sources such as coffee and fruit processing wastes and other
agricultural and food waste. The efficient conversion of
lowcost substrates to advanced biofuels requires development of
improved microbial catalysts (Hughes and Riedmuller 2014;
Koutinas et al. 2014; Peralta-Yahya et al. 2012). Economic
feasibility of biosynthetic fuel and chemical production depends on
optimization of these biocatalysts to achieve high yields of the
desired products. Saccharomyces cerevisiae is currently the
most employed microbial catalyst in the biotechnology industry,
but this yeast is limited in its range of substrates for producing
fuel ethanol, and although genetic engineering has improved its
utilization of the constituent pentose sugars of lignocellulosic
materials, development of a recombinant S. cerevisiae strain
capable of efficient pentose utilization remains a challenge
(Casey et al. 2013; Garcia Sanchez et al. 2010; Hughes et al.
2009a, b; Kim et al. 2013a, b; Matsushika et al. 2014; Nielsen
et al. 2013; Oreb et al. 2012; Zhou et al. 2012). Other microbial
catalysts are being investigated for the production of biofuels
and value-added bioproducts. One cand (...truncated)