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Irradiation of Yarrowia lipolytica NRRL YB-567 creating novel strains with enhanced ammonia and oil production on protein and carbohydrate substrates.

Lindquist MR, López-Núñez JC, Jones MA, Cox EJ, Pinkelman RJ, Bang SS, Moser BR, Jackson MA, Iten LB, Kurtzman CP, Bischoff KM, Liu S, Qureshi N, Tasaki K, Rich JO, Cotta MA, Saha BC, Hughes SR - Appl. Microbiol. Biotechnol. (2015)

Bottom Line: 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.These mutant strains will be investigated further for potential application in commercial biodiesel production.

View Article: PubMed Central - PubMed

Affiliation: 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.

ABSTRACT
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 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.

No MeSH data available.


Related in: MedlinePlus

Scanning electron micrographs of Yarrowia lipolytica mutant strains A, B, B2, C, D, E, and F compared to wild-type strain (Yarrowia lipolytica NRRL YB-567) grown on LNG medium. Scale is indicated by a bar in the lower left corner of each micrograph
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Fig2: Scanning electron micrographs of Yarrowia lipolytica mutant strains A, B, B2, C, D, E, and F compared to wild-type strain (Yarrowia lipolytica NRRL YB-567) grown on LNG medium. Scale is indicated by a bar in the lower left corner of each micrograph

Mentions: Scanning electron micrographs and light microscopy photographs (Figs. 2 and 3, respectively) show the morphological differences in the mutant strains when compared to the wild-type strain and to one another. The scanning electron micrographs (Fig. 2) show cells from Y. lipolytica wild-type and mutant strains in liquid LNG medium inoculated with a culture in LNG medium and incubated for 24 h at 30 °C and 100 rpm. The most notable morphological differences were the much smaller cell size (yeast-like forms about 1–2 μm in length) of mutant strains D, E, and F compared to the other mutant strains, A, B, B2, and C, and to the wild-type strain (yeast forms about 5 μm in length) and the bumpier cell surface of mutant strain F compared to the other strains. The cells of the wild-type strain exhibited both yeast (mostly ellipsoidal) and pseudohyphal forms with the yeast forms similar in size and shape. The surfaces of the yeast forms were smoother than the pseudohyphal projections, and the cells had budding scars at the polar ends. Cells of mutant strain A varied widely in size and shape; mostly, yeast and pseudohyphal forms with budding scars at the ends were present, although a few somewhat distorted shapes were also seen. The surfaces of most cells were very slightly bumpy, although a few of the yeast forms had relatively smooth surfaces. The cells of mutant strain B consisted of numerous very elongated forms and a few other distorted shapes. Both yeast and pseudohyphal forms were present, and some smooth and some bumpy surfaces were observed for all forms. The cells of mutant strain B2 had the largest variety of shapes and sizes, including elongated, distorted, yeast, and pseudohyphal forms, mostly smooth, but also some with slightly bumpy and a few with very bumpy surfaces. Cells of mutant strain C were mainly yeast and pseudohyphal forms with a few distorted elongated forms; almost all have bumpy surfaces. Cells of mutant strains D and F showed extensive branching and were intertwined forming a layer-like assembly; the surfaces of all cells were very bumpy. It was difficult to distinguish individual yeast and pseudohyphal forms, although both seemed to be present. The cells of mutant strain E were all misshapen yeast and pseudohyphal forms with extremely irregular surfaces.Fig. 2


Irradiation of Yarrowia lipolytica NRRL YB-567 creating novel strains with enhanced ammonia and oil production on protein and carbohydrate substrates.

Lindquist MR, López-Núñez JC, Jones MA, Cox EJ, Pinkelman RJ, Bang SS, Moser BR, Jackson MA, Iten LB, Kurtzman CP, Bischoff KM, Liu S, Qureshi N, Tasaki K, Rich JO, Cotta MA, Saha BC, Hughes SR - Appl. Microbiol. Biotechnol. (2015)

Scanning electron micrographs of Yarrowia lipolytica mutant strains A, B, B2, C, D, E, and F compared to wild-type strain (Yarrowia lipolytica NRRL YB-567) grown on LNG medium. Scale is indicated by a bar in the lower left corner of each micrograph
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4628078&req=5

Fig2: Scanning electron micrographs of Yarrowia lipolytica mutant strains A, B, B2, C, D, E, and F compared to wild-type strain (Yarrowia lipolytica NRRL YB-567) grown on LNG medium. Scale is indicated by a bar in the lower left corner of each micrograph
Mentions: Scanning electron micrographs and light microscopy photographs (Figs. 2 and 3, respectively) show the morphological differences in the mutant strains when compared to the wild-type strain and to one another. The scanning electron micrographs (Fig. 2) show cells from Y. lipolytica wild-type and mutant strains in liquid LNG medium inoculated with a culture in LNG medium and incubated for 24 h at 30 °C and 100 rpm. The most notable morphological differences were the much smaller cell size (yeast-like forms about 1–2 μm in length) of mutant strains D, E, and F compared to the other mutant strains, A, B, B2, and C, and to the wild-type strain (yeast forms about 5 μm in length) and the bumpier cell surface of mutant strain F compared to the other strains. The cells of the wild-type strain exhibited both yeast (mostly ellipsoidal) and pseudohyphal forms with the yeast forms similar in size and shape. The surfaces of the yeast forms were smoother than the pseudohyphal projections, and the cells had budding scars at the polar ends. Cells of mutant strain A varied widely in size and shape; mostly, yeast and pseudohyphal forms with budding scars at the ends were present, although a few somewhat distorted shapes were also seen. The surfaces of most cells were very slightly bumpy, although a few of the yeast forms had relatively smooth surfaces. The cells of mutant strain B consisted of numerous very elongated forms and a few other distorted shapes. Both yeast and pseudohyphal forms were present, and some smooth and some bumpy surfaces were observed for all forms. The cells of mutant strain B2 had the largest variety of shapes and sizes, including elongated, distorted, yeast, and pseudohyphal forms, mostly smooth, but also some with slightly bumpy and a few with very bumpy surfaces. Cells of mutant strain C were mainly yeast and pseudohyphal forms with a few distorted elongated forms; almost all have bumpy surfaces. Cells of mutant strains D and F showed extensive branching and were intertwined forming a layer-like assembly; the surfaces of all cells were very bumpy. It was difficult to distinguish individual yeast and pseudohyphal forms, although both seemed to be present. The cells of mutant strain E were all misshapen yeast and pseudohyphal forms with extremely irregular surfaces.Fig. 2

Bottom Line: 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.These mutant strains will be investigated further for potential application in commercial biodiesel production.

View Article: PubMed Central - PubMed

Affiliation: 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.

ABSTRACT
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 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.

No MeSH data available.


Related in: MedlinePlus