Lubuta, Patrice

Workman, Mhairi

With current global focus on renewable and sustainable technologies, there is a strong need to develop innovative solutions in industrial biotechnology. The non-conventional yeast Yarrowia lipolytica has considerable potential as a versatile cell factory, especially when compared to the typically applied organisms such as Saccharomyces cerevisiae which utilizes glycerol very slowly and requires genetic engineering for conversion of lignocellulosic sugar monomers xylose and arabinose. In this study we could show that Y. lipolytica IBT 446 is versatile and interesting as a biorefinery cell factory, as it is natural capable to utilize sugars typically found in plant hydrolysates (glucose, xylose, arabinose) together with glycerol in mixed carbon cultivations. Furthermore the strains W29 and H222 are naturally capable of conversion of glycerol, glucose and xylose

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Downloaded from orbit.dtu.dk on: Nov 08, 2017Characterizing Yarrowia lipolytica as a cell factory for sustainable bioprocessesLubuta, Patrice; Workman, MhairiPublication date:2016Document VersionPublisher's PDF, also known as Version of recordLink back to DTU OrbitCitation (APA):Lubuta, P., & Workman, M. (2016). Characterizing Yarrowia lipolytica as a cell factory for sustainablebioprocesses. Poster session presented at 12th International Conference on Renewable Resources andBiorefineries, Ghent, Belgium.Set up: Bioreactor batch cultivation, minimal media with 0,163 cmol L-1 (≈ 5 g L-1) of each carbon source. Characterizing Yarrowia lipolytica as a cell factory for sustainable bioprocesses Presented by Patrice Lubuta at Technical University of Denmark Email: palu@bio.dtu.dk Patrice Lubuta and Mhairi Workman  Fungal Physiology and Biotechnology Group Department of Systems Biology, Technical University of Denmark,  Building 223, 2800 Kgs. Lyngby, Denmark Background The utilization of an increasing amount of agro-waste and renewable low-cost substrates will be an ongoing challenge for the bio-based economy. Mobilization of these feedstocks is necessary for realization of biosustainability on an industrial scale. Cell factories traditionally applied In biotechnology are using a limited range of substrates (mainly glucose). However, a wider diversity in substrate range is highly desirable in developing biorefinery scenarios where seasonal variation in availability and composition of biomass feed-stocks typically appear.     Aim of the study This study evaluates the applicability of the non-conventional yeast Y. lipolytica as a cell factory for the conversion of glycerol and the lignocellulose hydrolysate sugars glucose, xylose and arabinose. A quantitative physiological investigation was carried out in order to assess the cellular performance of three different Y. ͏͏lipolytica wild type strains (IBT 446, W29 and H222) in single and mixed carbon source cultivations.  Summary and Conclusions With current global focus on renewable and sustainable technologies, there is a strong need to develop innovative solutions in industrial biotechnology. The non-conventional yeast Yarrowia lipolytica has considerable potential as a versatile cell factory, especially when compared to the typically applied organisms such as Saccharomyces cerevisiae which utilizes glycerol very slowly and requires genetic engineering for conversion of lignocellulosic sugar monomers xylose and arabinose. In this study we could show that Y. lipolytica IBT 446 is versatile and interesting as a biorefinery cell factory, as it is natural capable to utilize sugars typically found in plant hydrolysates (glucose, xylose, arabinose) together with glycerol in mixed carbon cultivations. Furthermore the strains W29 and H222 are naturally capable of conversion of glycerol, glucose and xylose.  Benchmarking strains on single carbon substrate  IBT 446 W29 H222 Glucose1 Glycerol1 Glucose Glycerol Glucose Glycerol Growth rate μmax (h-1) 0,24 ± 0,01 0,30 ± 0,01 0.28 ± 0.02 0.31 ± 0.02 0.28 ± 0.01 0.35 ± 0.01 Yield coefficients Ysx (cmol cmol-1) 0,69 ± 0,03 0,61 ± 0,01 0,67 ± 0,03 0,73 ± 0,01 0,64 ± 0,02 0.72 ± 0,01 Ysc (cmol cmol-1) 0,30 ± 0,01 0,23 ± 0,00 0,34 ± 0,02 0,30 ± 0,01 0,35 ± 0,004 0,21 ± 0,01 Ysm (cmol cmol-1) N/A* 0,17 ± 0,00 N/A* 0,015 ± 0,01 N/A* N/A** Total 0,99 ± 0,03 1,01 ± 0,01 1,02 ± 0,05 1,04 ± 0,01 0,98 ± 0,01 0,94 ± 0,01 1Workman 2013,  N/A: Not applicable: *below detection limit. **Carbon source not depleted at last sample point. Glucose vs Glycerol Growth experiments in shake flasks and defined minimal media with 0.65 cmol L-1 (≈ 20 g L-1) of the carbon source and cultivation time of 140 h: None of the strains showed growth on xylose or arabinose as the sole carbon source (verified by OD and HPLC measurements). Xylose and Arabinose IBT 446 - mixed substrates   Results:  Uptake cascade: 1. glycerol, 2. glucose, 3. xylose, 4. arabinose  Xylitol production and re-utilization  Growth on glycerol and glucose (primary carbon sources)  Maintenance of biomass after depletion of primary carbon sources W29 - mixed substrates   Results:  Uptake cascade: 1. glycerol, 2. glucose, 3. xylose (half consumed)  Co-consumption glycerol/glucose  Arabinose not consumed  Xylitol production but no re-utilization  Growth on glycerol and glucose (primary carbon sources)  Drop of biomass after depletion of primary carbon sources W29 - mixed substrates   Results:  Uptake cascade: 1. glycerol, 2. glucose, 3. xylose (half consumed)  Arabinose not consumed  Xylitol production but no re-utilization  Growth on glycerol and glucose (primary carbon sources)  Drop of biomass after depletion of primary carbon sources 

Characterizing <i>Yarrowia lipolytica</i> as a cell factory for sustainable bioprocesses

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Characterizing <i>Yarrowia lipolytica</i> as a cell factory for sustainable bioprocesses

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