scholarly journals Valorization of Brewers’ Spent Grain for the Production of Lipids by Oleaginous Yeast

Molecules ◽  
2018 ◽  
Vol 23 (12) ◽  
pp. 3052 ◽  
Author(s):  
Alok Patel ◽  
Fabio Mikes ◽  
Saskja Bühler ◽  
Leonidas Matsakas
Keyword(s):  
Lipid Content ◽  
Oleaginous Yeast ◽  
Lipid Production ◽  
Enzymatic Saccharification ◽  
Dry Weight ◽  
Lipid Concentration ◽  
Microwave Assisted ◽  
Spent Grain ◽  
Synthetic Media ◽  
By Products

Brewers’ spent grain (BSG) accounts for 85% of the total amount of by-products generated by the brewing industries. BSG is a lignocellulosic biomass that is rich in proteins, lipids, minerals, and vitamins. In the present study, BSG was subjected to pretreatment by two different methods (microwave assisted alkaline pretreatment and organosolv) and was evaluated for the liberation of glucose and xylose during enzymatic saccharification trials. The highest amount of glucose (46.45 ± 1.43 g/L) and xylose (25.15 ± 1.36 g/L) were observed after enzymatic saccharification of the organosolv pretreated BSG. The glucose and xylose yield for the microwave assisted alkaline pretreated BSG were 34.86 ± 1.27 g/L and 16.54 ± 2.1 g/L, respectively. The hydrolysates from the organosolv pretreated BSG were used as substrate for the cultivation of the oleaginous yeast Rhodosporidium toruloides, aiming to produce microbial lipids. The yeast synthesized as high as 18.44 ± 0.96 g/L of cell dry weight and 10.41 ± 0.34 g/L lipids (lipid content of 56.45 ± 0.76%) when cultivated on BSG hydrolysate with a C/N ratio of 500. The cell dry weight, total lipid concentration and lipid content were higher compared to the results obtained when grown on synthetic media containing glucose, xylose or mixture of glucose and xylose. To the best of our knowledge, this is the first report using hydrolysates of organosolv pretreated BSG for the growth and lipid production of oleaginous yeast in literature. The lipid profile of this oleaginous yeast showed similar fatty acid contents to vegetable oils, which can result in good biodiesel properties of the produced biodiesel.

scholarly journals Engineering cytoplasmic acetyl-CoA synthesis decouples lipid production from nitrogen starvation in the oleaginous yeast Rhodosporidium azoricum

2019 ◽  
Vol 18 (1) ◽  
Author(s):  
Silvia Donzella ◽  
Daniela Cucchetti ◽  
Claudia Capusoni ◽  
Aurora Rizzi ◽  
Silvia Galafassi ◽  
...  
Keyword(s):  
Lipid Content ◽  
Nitrogen Starvation ◽  
Neutral Lipids ◽  
Lipid Production ◽  
Industrial Process ◽  
Wild Type ◽  
Lipid Concentration ◽  
Atp Citrate Lyase ◽  
Acetyl Coa ◽  
Engineered Strain

Abstract Background Oleaginous yeasts are able to accumulate very high levels of neutral lipids especially under condition of excess of carbon and nitrogen limitation (medium with high C/N ratio). This makes necessary the use of two-steps processes in order to achieve high level of biomass and lipid. To simplify the process, the decoupling of lipid synthesis from nitrogen starvation, by establishing a cytosolic acetyl-CoA formation pathway alternative to the one catalysed by ATP-citrate lyase, can be useful. Results In this work, we introduced a new cytoplasmic route for acetyl-CoA (AcCoA) formation in Rhodosporidium azoricum by overexpressing genes encoding for homologous phosphoketolase (Xfpk) and heterologous phosphotransacetylase (Pta). The engineered strain PTAPK4 exhibits higher lipid content and produces higher lipid concentration than the wild type strain when it was cultivated in media containing different C/N ratios. In a bioreactor process performed on glucose/xylose mixture, to simulate an industrial process for lipid production from lignocellulosic materials, we obtained an increase of 89% in final lipid concentration by the engineered strain in comparison to the wild type. This indicates that the transformed strain can produce higher cellular biomass with a high lipid content than the wild type. The transformed strain furthermore evidenced the advantage over the wild type in performing this process, being the lipid yields 0.13 and 0.05, respectively. Conclusion Our results show that the overexpression of homologous Xfpk and heterologous Pta activities in R. azoricum creates a new cytosolic AcCoA supply that decouples lipid production from nitrogen starvation. This metabolic modification allows improving lipid production in cultural conditions that can be suitable for the development of industrial bioprocesses using lignocellulosic hydrolysates.

scholarly journals Exogenous l-proline improved Rhodosporidium toruloides lipid production on crude glycerol

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Rasool Kamal ◽  
Yuxue Liu ◽  
Qiang Li ◽  
Qitian Huang ◽  
Qian Wang ◽  
...  
Keyword(s):  
Cell Growth ◽  
Lipid Content ◽  
Crude Glycerol ◽  
Oleaginous Yeast ◽  
Lipid Production ◽  
Rhodosporidium Toruloides ◽  
Biodiesel Production ◽  
Lipid Yield ◽  
Stress Agent ◽  
Glycerol Consumption

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.

scholarly journals Distribution and isolation of microalgae for lipid production in selected freshwater reservoirs of northern Thailand

2018 ◽  
Vol 19 (1) ◽  
pp. 343-350
Author(s):  
TIPPAWAN PRASERTSIN ◽  
YUWADEE PEERAPORNPISAL
Keyword(s):  
Water Quality ◽  
Lipid Content ◽  
Trophic Status ◽  
Lipid Production ◽  
Dry Weight ◽  
Botryococcus Braunii ◽  
Chemical Parameters ◽  
Northern Thailand ◽  
Surrounding Areas ◽  
Physical And Chemical

Prasertsin T, Peerapornpisal Y. 2018. Distribution and isolation of microalgae for lipid production in selected freshwater reservoirs of northern Thailand. Biodiversitas 19: 343-350. Nong Bau Reservoir and Chiang Saen Lake are considered important freshwater reservoirs of Chiang Rai Province located in northern Thailand. The surrounding areas of these water bodies are host to a range of human activities that influence water quality. Moreover, to date, there are have not been any studies on the water quality and the distribution of microalgae in these places. The physical and chemical parameters of the water quality and microalgae were carried out in Nong Bua Reservoir and Chiang Saen Lake during the months of May, July, and October of 2015. Microalgae were isolated in order to investigate the lipid-producing abilities. Samples collected from Nong Bua Reservoir have revealed the presence of seven divisions, 90 species of algae. Six divisions, 55 species of algae were found in Chiang Saen Lake. The trophic status of the water was evaluated from the main parameters (AARL-PC Score), and it was determined that Nong Bua Reservoir was of meso-eutrophic status and Chiang Saen Lake was of mesotrophic status. Microalgae were isolated from Nong Bua Reservoir and Chiang Saen Lake for the purposes of studying lipid content; 25 and 6 isolations were identified, respectively. The lipid content was highest in Botryococcus braunii (39.25 ± 0.32% dry weight) followed by Ankistrodesmus sp. (26.80 ± 0.44% dry weight) and Coelastrum microsporum (24.95 ± 0.55% dry weight). The lowest lipid content was found in Planktolyngbya sp. (7.25 ± 0.43% dry weight). The indigenous strains of microalgae of Nong Bau Reservoir and Chiang Saen Lake’s can be considered quite promising as model strains in terms of the production of biofuel within the country.

Lipid metabolism and cell composition of the oleaginous yeast Apiotrichum curvatum grown at different carbon to nitrogen ratios

1990 ◽  
Vol 36 (5) ◽  
pp. 318-326 ◽  
Author(s):  
Wan-Soo Park ◽  
Patricia A. Murphy ◽  
Bonita A. Glatz
Keyword(s):  
Lipid Content ◽  
Lipid Accumulation ◽  
Catalase Activity ◽  
Oleaginous Yeast ◽  
Specific Activity ◽  
Dry Weight ◽  
Marker Enzyme ◽  
Cell Composition ◽  
Endogenous Lipid ◽  
Apiotrichum Curvatum

Apiotrichum curvatum ATCC 20509, an oleaginous yeast that can accumulate up to 60% of its cellular dry weight as intracellular lipid when grown with excess carbon, was grown in nitrogen-limited, balanced, and lactose-free medium with asparagine as nitrogen source and lactose as carbon source. Biomass and lipid accumulation were measured, cell composition was analyzed, and catalase activity was followed as marker enzyme for peroxisomes. The organism accumulated 54% of its dry weight as total cellular lipid when grown under nitrogen limitation and accumulated only 20–25% of its dry weight as lipid when grown in balanced medium. When starved for carbon, cells utilized endogenous lipid and carbohydrate as carbon and energy sources; the intracellular contents of lipid and carbohydrate decreased by 31 and 26%, respectively. Intracellular carbohydrates also seemed to be used as intermediates for lipid accumulation and lipid turnover. Catalase activity was strongly induced (over 10-fold increase in specific activity) when cells metabolized endogenous lipid. The lipid content of cells was inversely related to catalase activity and to intracellular protein or total nitrogen content. Lipid content showed no correlation with intracellular carbohydrate content. Key words: lipid, peroxisomes, oleaginous, Apiotrichum curvatum.

scholarly journals Enhanced inhibitor tolerance and increased lipid productivity through adaptive laboratory evolution in the oleaginous yeast Metshnikowia pulcherrima

2020 ◽  
Author(s):  
Robert H. Hicks ◽  
Yuxin Sze ◽  
Christopher J. Chuck ◽  
Daniel A. Henk
Keyword(s):  
Formic Acid ◽  
Oleaginous Yeast ◽  
Lipid Production ◽  
Dry Weight ◽  
Lipid Productivity ◽  
Adaptive Laboratory Evolution ◽  
Stirred Tank Bioreactor ◽  
Inhibitor Tolerance ◽  
Laboratory Evolution ◽  
Wide Range

AbstractMicrobial lipid production from second generation feedstocks presents a sustainable route to future fuels, foods and bulk chemicals. The oleaginous yeast Metshnikowia pulcherrima has previously been investigated as a potential platform organism for lipid production due to its ability to be grown in non-sterile conditions and metabolising a wide range of oligo- and monosaccharide carbon sources within lignocellulosic hydrolysates. However, the generation of inhibitors from depolymerisation causes downstream bioprocessing complications, and despite M. pulcherrima’s comparative tolerance, their presence is deleterious to both biomass and lipid formation. Using either a single inhibitor (formic acid) or an inhibitor cocktail (formic acid, acetic acid, fufural and HMF), two strategies of adaptive laboratory evolution were performed to improve M. pulcherrima’s fermentation inhibitor tolerance. Using a sequential batch culturing approach, the resulting strains from both strategies had increased growth rates and reduced lag times under inhibiting conditions versus the progenitor. Interestingly, the lipid production of the inhibitor cocktail evolved strains markedly increased, with one strain producing 41% lipid by dry weight compared to 22% of the progenitor. The evolved species was cultured in a non-sterile 2L stirred tank bioreactor and accumulated lipid rapidly, yielding 6.1 g/L of lipid (35% cell dry weight) within 48 hours; a lipid productivity of 0.128 g L-1 h-1. Furthermore, the lipid profile was analogous to palm oil, consisting of 39% C16:0 and 56% C18:1 after 48 hours.

scholarly journals Cutaneotrichosporon oleaginosus: A Versatile Whole-Cell Biocatalyst for the Production of Single-Cell Oil from Agro-Industrial Wastes

Catalysts ◽  
2021 ◽  
Vol 11 (11) ◽  
pp. 1291
Author(s):  
Nicola Di Fidio ◽  
Filippo Minonne ◽  
Claudia Antonetti ◽  
Anna Maria Raspolli Galletti
Keyword(s):  
Single Cell ◽  
Oleaginous Yeast ◽  
State Of The Art ◽  
Lipid Production ◽  
Industrial Applications ◽  
Industrial Wastes ◽  
Single Cell Oil ◽  
Whole Cell ◽  
Whole Cell Biocatalyst ◽  
By Products

Cutaneotrichosporon oleaginosus is an oleaginous yeast with several favourable qualities: It is fast growing, accumulates high amounts of lipids and has a very broad substrate spectrum. Its resistance to hydrolysis by-products makes it a promising biocatalyst for custom tailored microbial oils. C. oleaginosus can accumulate up to 60 wt.% of its biomass as lipids. This species is able to grow by using several compounds as a substrate, such as acetic acid, biodiesel-derived glycerol, N-acetylglucosamine, lignocellulosic hydrolysates, wastepaper and other agro-industrial wastes. This review is focused on state-of-the-art innovative and sustainable biorefinery schemes involving this promising yeast and second- and third-generation biomasses. Moreover, this review offers a comprehensive and updated summary of process strategies, biomass pretreatments and fermentation conditions for enhancing lipid production by C. oleaginosus as a whole-cell biocatalyst. Finally, an overview of the main industrial applications of single-cell oil is reported together with future perspectives.

BIOASSAY OF PROLACTIN

1969 ◽  
Vol 60 (1) ◽  
pp. 91-100 ◽  
Author(s):  
Charles S. Nicoll
Keyword(s):  
Epithelial Cells ◽  
Lipid Content ◽  
Dry Weight ◽  
Fat Content ◽  
Assay Sensitivity ◽  
Mucosal Epithelium ◽  
Wet Weight ◽  
Total Dry Weight

ABSTRACT The response of the pigeon crop-sac to systemically acting prolactin (injected subcutaneously) was evaluated by measuring the wet weight of the responsive lateral lobes of the organ and by determining the dry weight of a 4 cm diameter disc of mucosal epithelium taken from one hemicrop. Of several different injection schedules tested, administration of prolactin in four daily injections was found to yield optimal responses. When compared with a graded series of prolactin doses, measurement of the mucosal dry weight proved to be a better method of response quantification than determination of the crop-sac wet weight with respect to both assay sensitivity and precision. The submucosal tissue of the crop-sac was estimated to constitute about 64 % of the total dry weight of the unstimulated organ and it was found to be relatively unresponsive to prolactin stimulation in comparison with the mucosa. The lipid content of the mucosal epithelium was determined using unstimulated crop-sacs or tissues which showed varying degrees of prolactin-induced proliferation. The fat content of the mucosal epithelial cells increased only slightly more rapidly than the dry weight or the defatted dry weight of the mucosa. Suggestions are made for the further improvement of the systemic crop-sac assay for prolactin.

Regioselective Synthesis of Potent 4,5,6,7-Tetrahydroindazole Derivatives via Microwave-assisted Vilsmeier-Haack Reaction and their Antioxidant Activity Evaluation

2019 ◽  
Vol 16 (3) ◽  
pp. 194-201 ◽  
Author(s):  
Renu Bala ◽  
Vandana Devi ◽  
Pratibha Singh ◽  
Navjot Kaur ◽  
Pawandeep Kaur ◽  
...  
Keyword(s):  
Antioxidant Activity ◽  
Microwave Irradiation ◽  
Biological Activities ◽  
Reaction Times ◽  
Conventional Heating ◽  
High Chemical ◽  
Microwave Assisted ◽  
Work Up ◽  
Active Methylene ◽  
By Products

Background: Tetrahydroindazole, a member of the fused-pyrazole system, is a least studied class of heterocyclic compounds owing to its scarcity in nature. However, a large number of synthetically prepared tetrahydroindazoles are known to show a variety of biological activities such as interleukin- 2 inducible T-Cell kinase inhibitors, AMPA receptor positive allosteric modulators, antitumor, antituberculosis, anti-inflammatory and antimicrobial activities. Vilsmeier-Haack reaction is one of the most important chemical reactions used for formylation of electron rich arenes. Even though Vilsmeier- Haack reaction was studied on a wide variety of hydrazones derived from active methylene compounds, literature lacks the examples of the use of 4-substituted cyclohexanones as a substrate for the synthesis of 4,5,6,7-tetrahydroindazoles. The study of the reaction of Vilsmeier-Haack reagent with hydrazones derived from cyclic keto compounds having active methylene has been considered the interested topic of investigation. In the present study, ethyl cyclohexanone-4-carboxylate was treated with one equivalent of various hydrazines for two hours and the resulted hydrazones were further treated with an OPC-VH reagent (Vilsmeier-Haack reagent isolated from phthaloyl dichloride and N,Ndimethylformamide) afforded 4,5,6,7-tetrahydroindazoles in excellent yields. The synthesized compounds 4a-f and 5a-f were screened for their antioxidant activities using the DPPH radical scavenging assay. The target compounds were synthesized regioselectively using 4+1 approach in excellent yields. A number of experiments using both conventional heating as well as microwave irradiation methods were tried and on comparison, microwave irradiation method was found excellent in terms of easy work up, high chemical yields, shortened reaction times, clean and, no by-products formation. Some of the synthesized compounds showed significant antioxidant activity. The microwave assisted synthesis of 4,5,6,7-tetrahydroindazoles from ethyl cyclohexanone-4-carboxylate has been reported under mild conditions in excellent yield. Easy work up, high chemical yield, shortened reaction times, clean and no by-products formation are the major advantages of this protocol. These advantages may make this method useful for chemists who are interested in developing novel 4,5,6,7-tetrahydroindazole based drugs.

scholarly journals Valorization of sugarcane bagasse by chemical pretreatment and enzyme mediated deconstruction

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Vihang S. Thite ◽  
Anuradha S. Nerurkar
Keyword(s):  
Cell Wall ◽  
Sugarcane Bagasse ◽  
Structural Changes ◽  
Enzymatic Saccharification ◽  
Dry Weight ◽  
Gravimetric Analysis ◽  
Chemical Pretreatment ◽  
Cell Wall Degrading Enzymes ◽  
First Time ◽  
Soluble Components

Abstract After chemical pretreatment, improved amenability of agrowaste biomass for enzymatic saccharification needs an understanding of the effect exerted by pretreatments on biomass for enzymatic deconstruction. In present studies, NaOH, NH4OH and H2SO4 pretreatments effectively changed visible morphology imparting distinct fibrous appearance to sugarcane bagasse (SCB). Filtrate analysis after NaOH, NH4OH and H2SO4 pretreatments yielded release of soluble reducing sugars (SRS) in range of ~0.17–0.44%, ~0.38–0.75% and ~2.9–8.4% respectively. Gravimetric analysis of pretreated SCB (PSCB) biomass also revealed dry weight loss in range of ~25.8–44.8%, ~11.1–16.0% and ~28.3–38.0% by the three pretreatments in the same order. Release of soluble components other than SRS, majorly reported to be soluble lignins, were observed highest for NaOH followed by H2SO4 and NH4OH pretreatments. Decrease or absence of peaks attributed to lignin and loosened fibrous appearance of biomass during FTIR and SEM studies respectively further corroborated with our observations of lignin removal. Application of commercial cellulase increased raw SCB saccharification from 1.93% to 38.84%, 25.56% and 9.61% after NaOH, H2SO4 and NH4OH pretreatments. Structural changes brought by cell wall degrading enzymes were first time shown visually confirming the cell wall disintegration under brightfield, darkfield and fluorescence microscopy. The microscopic evidence and saccharification results proved that the chemical treatment valorized the SCB by making it amenable for enzymatic saccharification.

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