scholarly journals Lipid Production from Crude Glycerol by Newly Isolated Oleaginous Yeasts: Strain Selection, Molecular Identification and fatty acid analysis

Author(s):  
Derya Berikten ◽  
Emir Zafer Hosgun ◽  
Ayşe Gökdal Otuzbiroğlu ◽  
Berrin Bozan ◽  
Merih Kıvanç

Abstract Biodiesel is a renewable alternative fuel and glycerol as a main byproduct of the manufacturing process. Lipids could be produced from crude glycerol by using yeasts. The ability of 107 yeast strains to utilize glycerol was screened and 92 of these were selected. 60 strains were determined as a potential for lipid production by Sudan Black B staining. After secondary screening 25 of them showed specific growth rates (OD 600), high biomass production and lipid content. These strains were identified as Pichia cactophila, P. fermentans, P. anomala, Rhodotorula mucilaginosa, R. dairenensis, Clavispora lusitaniae, Saccharomyces cerevisiae, Wickerhamomyces anomalus, Candida glabrata, C. inconspicua, C. albicans, Yarrowia lipolytica with molecular identifications based on ITS and D1/D2 26S rDNA sequences. The results showed that P. cactophila accumulated lipid up to 64.94%, the highest lipid content. C16:0, C18:0, C18:1 and C18:2 essential fatty acids for biodiesel production were detected by GC-MS in the lipids accumulated by all strains. P. cactophila and C. lusitaniae were reported for the first time as lipid-producing yeasts. The results suggest that selected 25 isolates have the ability to grow on crude glycerol and especially P. cactophila produce lipid that has potential use as a feedstock for second generation biodiesel production.

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Rasool Kamal ◽  
Yuxue Liu ◽  
Qiang Li ◽  
Qitian Huang ◽  
Qian Wang ◽  
...  

Abstract Background Crude glycerol as a promising feedstock for microbial lipid production contains several impurities that make it toxic stress inducer at high amount. Under stress conditions, microorganisms can accumulate l-proline as a safeguard. Herein, l-proline was assessed as an anti-stress agent in crude glycerol media. Results Crude glycerol was converted to microbial lipids by the oleaginous yeast Rhodosporidium toruloides CGMCC 2.1389 in a two-staged culture mode. The media was supplied with exogenous l-proline to improve lipid production efficiency in high crude glycerol stress. An optimal amount of 0.5 g/L l-proline increased lipid titer and lipid yield by 34% and 28%, respectively. The lipid titer of 12.2 g/L and lipid content of 64.5% with a highest lipid yield of 0.26 g/g were achieved with l-proline addition, which were far higher than those of the control, i.e., lipid titer of 9.1 g/L, lipid content of 58% and lipid yield of 0.21 g/g. Similarly, l-proline also improved cell growth and glycerol consumption. Moreover, fatty acid compositional profiles of the lipid products was found suitable as a potential feedstock for biodiesel production. Conclusion Our study suggested that exogenous l-proline improved cell growth and lipid production on crude glycerol by R. toruloides. The fact that higher lipid yield as well as glycerol consumption indicated that l-proline might act as a potential anti-stress agent for the oleaginous yeast strain.


2020 ◽  
Author(s):  
Francesca Martani ◽  
Letizia Maestroni ◽  
Mattia Torchio ◽  
Diletta Ami ◽  
Antonino Natalello ◽  
...  

Abstract BackgroundLipids from oleaginous yeasts emerged as a sustainable alternative to vegetable oils and animal fat to produce biodiesel, the biodegradable and environmentally friendly counterpart of petro-diesel fuel. To develop economically viable microbial processes, the use of residual feedstocks as growth and production substrates is required.ResultsIn this work we investigated sugar beet pulp (SBP) and molasses, the main residues of sugar beet processing, as sustainable substrates for the growth and lipid accumulation by the oleaginous yeast Lipomyces starkeyi. We observed that in hydrolysed SBP the yeast cultures reached a limited biomass, cellular lipid content, lipid production and yield (2.5 g/L, 19.2 %, 0.5 g/L and 0.08 g/g, respectively). To increase the initial sugar availability, cells were grown in SBP blended with molasses. Under batch cultivation, the cellular lipid content was more than doubled (47.2 %) in the presence of 6 % molasses. Under pulsed-feeding cultivation, final biomass, cellular lipid content, lipid production and lipid yield were further improved, reaching respectively 20.5 g/L, 49.2 %, 9.7 g/L and 0.178 g/g. Finally, we observed that SBP can be used instead of ammonium sulphate to fulfil yeasts nitrogen requirement in molasses-based media for microbial oil production. ConclusionsThis study demonstrates for the first time that SBP and molasses can be blended to create a feedstock for the sustainable production of lipids by L. starkeyi. The data obtained pave the way to further improve lipid production by designing a fed-batch process in bioreactor.


2020 ◽  
Author(s):  
Francesca Martani ◽  
Letizia Maestroni ◽  
Mattia Torchio ◽  
Diletta Ami ◽  
Antonino Natalello ◽  
...  

Abstract Background: Lipids from oleaginous yeasts emerged as a sustainable alternative to vegetable oils and animal fat to produce biodiesel, the biodegradable and environmentally friendly counterpart of petro-diesel fuel. To develop economically viable microbial processes, the use of residual feedstocks as growth and production substrates is required.Results: In this work we investigated sugar beet pulp (SBP) and molasses, the main residues of sugar beet processing, as sustainable substrates for the growth and lipid accumulation by the oleaginous yeast Lipomyces starkeyi. We observed that in hydrolysed SBP the yeast cultures reached a limited biomass, cellular lipid content, lipid production and yield (2.5 g/L, 19.2 %, 0.5 g/L and 0.08 g/g, respectively). To increase the initial sugar availability, cells were grown in SBP blended with molasses. Under batch cultivation, the cellular lipid content was more than doubled (47.2 %) in the presence of 6 % molasses. Under pulsed-feeding cultivation, final biomass, cellular lipid content, lipid production and lipid yield were further improved, reaching respectively 20.5 g/L, 49.2 %, 9.7 g/L and 0.178 g/g. Finally, we observed that SBP can be used instead of ammonium sulphate to fulfil yeasts nitrogen requirement in molasses-based media for microbial oil production. Conclusions: This study demonstrates for the first time that SBP and molasses can be blended to create a feedstock for the sustainable production of lipids by L. starkeyi. The data obtained pave the way to further improve lipid production by designing a fed-batch process in bioreactor.


2019 ◽  
Author(s):  
Francesca Martani ◽  
Letizia Maestroni ◽  
Mattia Torchio ◽  
Diletta Ami ◽  
Antonino Natalello ◽  
...  

Abstract Background: Lipids from oleaginous yeasts emerged as sustainable alternative to vegetable oils and animal fat to produce biodiesel, the biodegradable and environmentally friendly counterpart of diesel fuel. To develop economically viable microbial processes, the use of residual feedstocks as growth substrates is required.Results: In this work, we investigated sugar beet pulp (SBP) and molasses, the main residues of sugar beet processing, as sustainable substrates for the growth and lipid accumulation by the oleaginous yeast Lipomyces starkeyi. Initially, we observed that in hydrolyzed SBP the yeast cultures reached a limited biomass, cellular lipid content, lipid production and yield (2.5 g/L, 19.2 %, 0.5 g/L and 0.08 g/g, respectively). Secondly, to increase the initial sugar availability, cells were grown in SBP blended with molasses. Under batch cultivation, the cellular lipid content was more than doubled (47.2 %) in the presence of 6 % molasses. Under pulsed-feeding cultivation, final biomass, cellular lipid content, lipid production and lipid yield were further improved, reaching respectively 20.5 g/L, 49.2 %, 9.7 g/L and 0.178 g/g. Finally, we observed that SBP can be used instead of ammonium sulfate to fulfill yeasts nitrogen requirement in molasses-based media for microbial oil production. Conclusions: This study demonstrated for the first time that SBP and molasses can be blended to create a feedstock for the sustainable production of lipids by L. starkeyi. The data obtained pave the way to further improve lipid production by designing a fed-batch cultivation mode in bioreactor.


2017 ◽  
Vol 5 (2) ◽  
pp. 88
Author(s):  
Bruno Loureiro Ladeira ◽  
Janaína Fernandes de Medeiros Burkert ◽  
Carlos André Burkert

Biodiesel production, which has been increasing worldwide, transformed crude glycerol, the main byproduct of the reaction, into a commodity of low commercial value, especially due to the high costs involved in its purification process and to the fact that the market cannot account for its generation. Therefore, this study aims at contributing to the search for technological alternatives to the use of surplus crude glycerol so as to add value to this byproduct by using it as the carbon source for the yeast Meyerozyma guilliermondii, in order to yield lipids. The Central Composite Rotational Design was proposed to establish empirical models, codified for lipid content, production and productivity as the result of concentrations of Magnesium Sulfate Heptahydrate (MgSO4.7H2O) and yeast extract. In the optimized conditions, the lipid content in dry basis, lipid production and lipid productivity were 8.25%, 1.10 g.L-1 and 0.0092 g.L-1.h-1, respectively obtained.


2021 ◽  
Vol 16 (10) ◽  
pp. 144-155
Author(s):  
Van Lal Michael Chhandama ◽  
Belur Kumudini Satyan

Microalgae emerged as a competent feedstock for biodiesel production because of high growth rate and lipid content. This work focuses on isolation of novel microalgal strain from different sources of water for the production of biodiesel. The isolated microalgae, Pleurastrum insigne possessed high lipid content (~28 % dcw), further optimized to 57.06 % dcw using a statistical design (CCD) under Response Surface Methodology. Lipid production was optimized by nutrient (nitrogen and phosphorus) and pH stress. The different type of fatty acids present in the optimized lipid was also profiled using GCMS. Biodiesel yield was found to be 82.14 % of the total lipid and the fuel properties tested have met IS, ASTM and EN biodiesel standards.


2020 ◽  
Vol 19 (1) ◽  
Author(s):  
Francesca Martani ◽  
Letizia Maestroni ◽  
Mattia Torchio ◽  
Diletta Ami ◽  
Antonino Natalello ◽  
...  

Abstract Background Lipids from oleaginous yeasts emerged as a sustainable alternative to vegetable oils and animal fat to produce biodiesel, the biodegradable and environmentally friendly counterpart of petro-diesel fuel. To develop economically viable microbial processes, the use of residual feedstocks as growth and production substrates is required. Results In this work we investigated sugar beet pulp (SBP) and molasses, the main residues of sugar beet processing, as sustainable substrates for the growth and lipid accumulation by the oleaginous yeast Lipomyces starkeyi. We observed that in hydrolysed SBP the yeast cultures reached a limited biomass, cellular lipid content, lipid production and yield (2.5 g/L, 19.2%, 0.5 g/L and 0.08 g/g, respectively). To increase the initial sugar availability, cells were grown in SBP blended with molasses. Under batch cultivation, the cellular lipid content was more than doubled (47.2%) in the presence of 6% molasses. Under pulsed-feeding cultivation, final biomass, cellular lipid content, lipid production and lipid yield were further improved, reaching respectively 20.5 g/L, 49.2%, 9.7 g/L and 0.178 g/g. Finally, we observed that SBP can be used instead of ammonium sulphate to fulfil yeasts nitrogen requirement in molasses-based media for microbial oil production. Conclusions This study demonstrates for the first time that SBP and molasses can be blended to create a feedstock for the sustainable production of lipids by L. starkeyi. The data obtained pave the way to further improve lipid production by designing a fed-batch process in bioreactor. Graphical abstract


2020 ◽  
Author(s):  
Nhung TT Pham ◽  
Maarten Reijnders ◽  
Maria Suarez-Diez ◽  
Bart Nijsse ◽  
Jan Springer ◽  
...  

Abstract Background: Cutaneotrichosporon oleaginosus ATCC 20509 is a fast growing oleaginous basidiomycete yeast that is able to grow in a wide range of low-cost carbon sources including crude glycerol, a byproduct of biodiesel production. When glycerol is used as a carbon source, this yeast can accumulate more than 50% lipids (w/w) with high concentrations of mono-unsaturated fatty acids.Results: To increase our understanding of this yeast and to provide a knowledge base for further industrial use, a FAIR re-annotated genome was used to build a genome-scale, constraint-based metabolic model containing 1553 reactions involving 1373 metabolites in 11 compartments. A new description of the biomass synthesis reaction was introduced to account for massive lipid accumulation in conditions with high carbon to nitrogen (C/N) ratio in the media. This condition-specific biomass objective function is shown to better predict conditions with high lipid accumulation using glucose, fructose, sucrose, xylose, and glycerol as sole carbon source.Conclusion: Contributing to the economic viability of biodiesel as renewable fuel, C. oleaginosus ATCC 20509 can effectively convert crude glycerol waste streams in lipids as a potential bioenergy source. Performance simulations are essential to identify optimal production conditions and to develop and fine tune a cost-effective production process. Our model suggests ATP-citrate lyase as a possible target to further improve lipid production.


Author(s):  
Jiali Gui ◽  
Shuangshuang Chen ◽  
Guiying Luo ◽  
Zixiang Wu ◽  
Yongxian Fan ◽  
...  

The lipid production potential of 8 microalgae species was investigated. Among these eight species, the best strain was a dominant bloom-causing dinoflagellate, Prorocentrum donghaiense ; this species had a lipid content of 49.32±1.99% and exhibited a lipid productivity of 95.47±0.99 mg L −1 d −1 , which was 2-fold higher than the corresponding values obtained for the oleaginous microalgae Nannochloropsis gaditana and Phaeodactylum tricornutum . P. donghaiense, which is enriched in C16:0 and C22:6, is appropriate for commercial DHA production. Nitrogen or phosphorus stress markedly induced lipid accumulation to levels surpassing 75% of the dry weight, increased the C18:0 and C17:1 contents, and decreased the C18:5 and C22:6 contents, and these effects resulted in decreases in the unsaturated fatty-acid levels and changes in the lipid properties of P. donghaiense such that the species met the biodiesel specification standards. Compared with the results obtained under N-deficient conditions, the enhancement in the activity of alkaline phosphatase of P. donghaiense observed under P-deficient conditions could partly alleviate the adverse effects on the photosynthetic system exerted by P deficiency to induce the production of more carbohydrates for lipogenesis. The supernatant of the algicidal bacterium Paracoccus sp. Y42 culture lysed P. donghaiense without decreasing its lipid content, which resulted in facilitation of the downstream oil extraction process and energy savings through the lysis of algal cells. The Y42 supernatant treatment improved the lipid profiles of algal cells by increasing their C16:0, C18:0 and C18:1 contents and decreasing their C18:5 and C22:6 contents, which is favourable for biodiesel production. IMPORTANCE This study demonstrates the high potential of P. donghaiense , a dominant bloom-causing dinoflagellate, for lipid production. Compared with previously studied oleaginous microalgae, P. donghaiense exhibit greater potential for practical application due to its higher biomass and lipid contents. Nutrient deficiency and the algicidal bacterium Paracoccus sp. Y42 could improve the suitability of the lipid profile of P. donghaiense for biodiesel production. Furthermore, Paracoccus sp. Y42 effectively lyse algal cells, which facilitates the downstream oil extraction process for biodiesel production and results in energy savings through the lysing of algal cells. This study provides a more promising candidate for the production of DHA for human nutritional products and of microalgal biofuel, as well as a more cost-effective method for breaking algal cells. The high lipid productivity of P. donghaiense and algal cell lysis by algicidal bacteria contribute to reductions in the production cost of microalgal oil.


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