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Poplar wood hemicellulose prehydrolysate was used for microbial oil production using an oleaginous microorganism Cryptococcus curvatus. Initially, the effect of substrate concentration and nitrogen content was investigated on synthetic media. Then poplar wood prehydrolysate without detoxification was used as substrate in the fermentation. The result showed that this strain is capable of consuming both hexose and pentose sugars, a challenge in fermentation of hemicellulosic streams. It was able to accumulate 36.98% of lipid and the fermentation resulted in 13.78 g/L of biomass and 5.13 g/L of lipid under optimum conditions after 164 h of fermentation. The lipid product obtained was characterized in terms of their fatty acid profiles. Overall, this study shows that it is possible to produce microbial oil from a sustainable renewable feedstock like poplar wood hemicellulose prehydrolysate. This robust strain used has the ability to grow on industrially produced hemicellulose which can help in the development of an integrated biorefinery, where all the three components of lignocellulosic biomass are utilized.

N-acetylglucosamine (GlcNAc), the monomeric constituent of chitin, is rarely studied for lipid production by oleaginous species. This study demonstrated that Cryptococcus curvatus had a great capacity to convert GlcNAc into lipid with high yield using a two-stage production process. Optimal inoculum age and inoculation size strongly improved the two-stage lipid production efficiency. More interestingly, this process rendered superior lipid production under non-sterile condition. The acetate liberated from GlcNAc was consumed timely, while the NH 4 + released was rarely assimilated. Lipid titre, lipid content and lipid yield reached 9.9 g/L, 56.9% and 0.23 g/g, respectively, which were significantly higher than those from the conventional process where cell growth and lipid accumulation were coupled. The resulting lipid samples had similar fatty acid compositional profiles to those of vegetable oil, suggesting their potential for biodiesel production. These findings strongly supported the two-stage process as an attractive strategy for better techno-economics of the chitin-to-biodiesel routes., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Tailoring lipids from oleaginous yeasts to contain specific types of fatty acid is of considerable interest to food, fuel, and pharmaceutical industries. In this study, the essential oil obtained from Citrus sinesus L. has been used to alter the fatty acid composition of two common oleaginous yeasts, Rhodosporidium toruloides and Cryptococcus curvatus. With increasing levels of essential oil in the medium, the metabolic flux of the fatty acid biosynthesis pathway shifted towards saturated fatty acid production. Essential oil reduced the activities of elongase and ∆9 desaturase. This made the lipid obtained from both these yeasts rich in saturated fatty acids. At certain specific concentrations of the essential oil in the medium, the lipid obtained from R. toruloides and C. curvatus cultures was similar to mahuwa butter and palm oil, respectively. Limonene is the major constituents of orange essential oil. Its effect on one of the oleaginous yeasts, R. toruloides, was also studied separately. Effects similar to orange essential oil were obtained with limonene. Thus, we can conclude that limonene in orange essential oil brings about compositional change of microbial lipid produced in this organism.

In the present study, synergistic effects were observed when glycerol was co-fermented with glucose and xylose for lipid production by the oleaginous yeast Cryptococcus curvatus. Glycerol was assimilated simultaneously with sugars at the beginning of the culture without adaption time. Furthermore, better lipid production results, i.e., lipid yield and lipid productivity of 18.0g/100g and 0.13g/L/h, respectively, were achieved when cells were cultured in blends of corn stover hydrolysates and biodiesel-derived glycerol than those in the hydrolysates alone. The lipid samples had fatty acid compositional profiles similar to those of vegetable oils, suggesting their potential for biodiesel production. This co-utilization strategy provides an extremely simple solution to advance lipid production from both lignocelluloses and biodiesel-derived glycerol in one step., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

The improvement of culture parameters for lipid production from acetate as carbon source was investigated using the oleaginous yeast Cryptococcus curvatus. A new pH regulation system dispensing acetate was developed for fed-batch culture and allowed obtaining nearly 80 g/L biomass within 60 h with a maximal growth rate of 0.28 h(-1). A biological model was developed from experimental data. The influence of three C/N ratios of 300, 500 and 900 were tested during a multi-phases process on lipid accumulation. The C/N ratio of 300 was reported to be the most suitable for lipid storage. No significant increase of lipids content was obtained with higher value. A maximal content of 60% DCW of lipid was obtained. The determination of fatty acids profiles of the microbial oils has confirmed that the valorization of acetate by microbial oils production was a promising perspective., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Microbial oils hold great potential as a suitable feedstock for the renewable production of biofuels. Specifically, the use of oleaginous yeasts offers several advantages related to cultivation and quality of lipid products. However, one of the major bottlenecks for large-scale production of yeast oils is found in the lipid extraction process. This work investigated the hydrothermal treatment of oleaginous yeast for hydrolysis and lipid extraction resulting in fatty acids used for biofuel production. The oleaginous yeast, Cryptococcus curvatus, was grown in 5 L bioreactors and the biomass slurry with 53±4% lipid content (dry weight basis) was treated at 280 °C for 1h with an initial pressure of 500 psi in batch stainless steel reactors. The hydrolysis product was separated and each of the resulting streams was further characterized. The hexane soluble fraction contained fatty acids from the hydrolysis of yeast triacylglycerides, and was low in nitrogen and minerals and could be directly integrated as feedstock into pyrolysis processing to produce biofuels. The proposed hydrothermal treatment addresses some current technological bottlenecks associated with traditional methodologies such as dewatering, oil extraction and co-product utilization. It also enhances the feasibility of using microbial biomass for production of renewable fuels and chemicals., (Copyright © 2014 Elsevier B.V. All rights reserved.)

A sequential hydrothermal liquefaction (SEQHTL) process was evaluated in this work for fractionating different component of yeast biomass. Sugar and protein were separated first at a lower temperature, and the remaining biomass was then converted to bio-oil at a higher temperature. The separated aqueous products were investigated to be recycled as a carbon and nitrogen sources for the yeast culture. In the first step of SEQHTL, the temperature effect on the yield of sugar/protein and inhibitory compounds (acetic acid and 5-hydroxymethyl furfural (5HMF)) was investigated. The highest yields of sugar and protein and a minimal level of inhibitory compounds were obtained at 180°C. At the second step of SEQHTL, the highest bio-oil yield was achieved at 240°C. In comparison to the one-step hydrothermal liquefaction process, SEQHTL produced a higher quality bio-oil with higher fatty acid and lower nitrogen contents., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

With its high content of carbohydrates and low percentage of lignin, corn fiber represents a renewable feedstock that can be processed to produce biofuels. Through a combination of pretreatment by lime and enzymatic hydrolysis, total reducing sugars of 700 mg/g corn fiber were released. This amount is equivalent to 92.7% of theoretically available sugars in corn fiber. The resulting hydrolysate itself did not support any growth of Cryptococcus curvatus. But with addition of minerals, C. curvatus grew to a cell density of 6.6 g/L in 6 days. Using the adapted cells, rapid sugar consumption and cell growth were observed. This study demonstrated that it is feasible to produce microbial lipids from corn fiber through pretreatment, enzymatic hydrolysis, and fermentation. In addition, C. curvatus is an excellent candidate for this application since it can utilize all major sugars, glucose, xylose, and arabinose with yield of cells and lipids as 0.55 and 0.27 g/g sugars, respectively., (© 2014 American Institute of Chemical Engineers.)

Cutaneotrichosporon oleaginosus ATCC 20509, previously known as Trichosporon oleaginosus, Cryptococcus curvatus, Apiotrichum curvatum or Candida curvata D is an oleaginous yeast with several favorable qualities: it is fast growing, accumulates high amounts of lipid and has a very broad substrate spectrum. Its resistance to hydrolysis byproducts and genetic accessibility make it a promising cell factory for custom tailored microbial oils. However, literature about this organism is of varying degree of quality. Moreover, due to numerous changes of the species name, reports are highly scattered and poorly cited. This led to a poor integration of the findings into a unified body of knowledge. Particularly, errors in strain name usage and consequently citation are found even in most recent literature. To simplify future work, this review provides an overview of published studies and main findings regarding the metabolic capacities of C. oleaginosus. [ABSTRACT FROM AUTHOR]

Waste paper was studied as a potential source for lipid production using the oleaginous yeast Cryptococcus curvatus for the first time. Three common types of waste paper, office paper, newspaper, and cardboard, were directly hydrolyzed by an enzyme cocktail to generate sugar-rich and nitrogen-limited hydrolysates. When these hydrolysates were used without any auxiliary nutrients by C. curvatus, the lipid content and lipid yield were higher than 50% and 200 mg/g, respectively. The nitrogen-rich enzyme cocktail exerted no negative effects on lipid production. Moreover, the integrated processes of enzymatic hydrolysis and lipid fermentation achieved comparable lipid yield to the separate hydrolysis and lipid production process. The resulting lipid samples had similar fatty acid compositional profiles to those of vegetable oils, which suggested their potential for biodiesel production. These findings strongly supported waste paper as appealing substrates for lipid production via oleaginous yeast, which provided cost-effective waste paper-to-lipids routes for sustainable biodiesel production. [ABSTRACT FROM AUTHOR]

In a world of finite resources, the transition to a sustainable post-fossil resources economy is essential. Within this concept, spent coffee grounds (SCGs) could be used for the production of bio-based chemicals, biopolymers and value-added products. Lipids and phenolics were initially extracted from SCGs. Alkaline pretreatment of the remaining SCGs solids has been optimised in this study via Central Composite Design to maximise lignin removal and overall sugar to polysaccharide conversion yield. The optimal SCGs pretreatment conditions were 0.06% (w/v) NaOH and 99.5 °C resulting in 35.7% lignin removal at 60 min pretreatment duration with low glucan and hemicellulose losses (ca. 2.5%). Enzymatic hydrolysis of the remaining SCGs alkaline pretreated solids resulted in 79.4% glucan, 74.5% mannan and 54.8% hemicellulose hydrolysis yield. The SCGs hydrolysate was used as fermentation feedstock in bioreactor cultures of Cryptococcus curvatus resulting in 36.1 g/L microbial oil with 0.25 g/g yield and 0.41 g/(L∙h) productivity. The total dry weight reached 62.7 g/L corresponding to 57.5% (w/w) intracellular lipid content. The microbial oil was rich in oleic acid (55.6%) followed by palmitic acid (18.9%) and stearic acid (14.0%). The proposed process led to the production of 7.81 g microbial oil per 100 g antioxidant- and oil-free SCGs. • Antioxidant- and oil-free SCGs were efficiently used for microbial oil production. • Optimal SCGs pretreatment conditions were 0.06% (w/v) NaOH and 99.5 °C. • Lignin removal of 35.7% with low glucan and hemicellulose losses was achieved. • Fermentation of SCGs hydrolysate led to 57.5% (w/w) intracellular lipid content. • 7.81 g microbial oil obtained per 100 g antioxidant and oil-free SCGs. [ABSTRACT FROM AUTHOR]

This study focuses on the use of acetic acid as carbon source for cell mass and lipid production using an oleaginous yeast, Cryptococcus curvatus . To improve this cell mass and lipid production on acetic acid, a numerical model was built to predict the yeast behavior. The model was first calibrated and validated on a fed-batch culture of the oleaginous yeast grown on acetic acid. After the validation step, this model was used as a predictive tool to search for optimal dilution rate and C/N ratios for continuous culture. A continuous culture was then launched using these culture parameters, revealing that the experimental data fitted the model predictions. At steady-state, cell mass concentration was stabilized at 26.7 g/L containing 48–53% of lipid content. A maximal lipid productivity of 0.54 g L −1 h −1 was obtained, which was superior to those reported in literature. This model could be qualified as a mechanistic one since it allows understanding reactions underlying the lipid production process. The experimental data fitted the model predictions, which confirmed that the use of a model is a powerful and predictive tool which could help the operator to process parameters to improve the culture. [ABSTRACT FROM AUTHOR]

Background: Acetic acid is routinely generated during lignocelluloses degradation, syngas fermentation, dark hydrogen fermentation and other anaerobic bioprocesses. Acetate stream is commonly regarded as a by-product and detrimental to microbial cell growth. Conversion of acetate into lipids by oleaginous yeasts may be a good choice to turn the by-product into treasure. Results: Ten well-known oleaginous yeasts were evaluated for lipid production on acetate under flask culture conditions. It was found that all of those yeasts could use acetate for microbial lipid production. In particular, Cryptococcus curvatus accumulated lipids up to 73.4 % of its dry cell mass weight. When the culture was held in a 3-L stirred-tank bioreactor, cell mass, lipid content, lipid yield and acetate consumption rate were 8.1 g/L, 49.9 %, 0.15 g/g and 0.64 g/L/h, respectively. The fatty acid compositional profiles of the acetate-derived lipids were similar to those of vegetable oil, suggesting their potential for biodiesel production. Continuous cultivation of C. curvatus was conducted under nitrogen-rich condition at a dilution rate of 0.04 h-1, the maximal lipid content and lipid yield were 56.7 % and 0.18 g/g, respectively. The specific lipid formation rate, lipid content and lipid yield were all higher under nitrogen-rich conditions than those obtained under nitrogen-limited conditions at the same dilution rates. Effective lipid production by C. curvatus was observed on corn stover hydrolysates containing 15.9 g/L acetate. Conclusions: Acetate is an effective carbon source for microbial lipid production by oleaginous yeasts. Continuous cultivation of C. curvatus on acetate was promising for lipid production under both nitrogen-rich and nitrogen-limited conditions. These results provide valuable information for developing and designing more efficient acetate-into-lipids bioprocess. [ABSTRACT FROM AUTHOR]

Cell disruption is an important step used to substantially increase lipid recovery prior to extraction. This is the first study exploring the effects of cell disruption methods on lipid recovery from different oleaginous microorganisms. Specifically, we examined the effects of cell disruption methods, including autoclaving, bead-beating, microwaving, sonication, acid digestion, and Soxhlet extraction on the yeast Cryptococcus curvatus, the fungus Mortierella isabellina and the microalga Chlorella sorokiniana. Although lipid recovery by cell disruption methods varied with microbial species, autoclaving and microwaving resulted in the lowest lipid contents for all tested strains. The Soxhlet method achieved the highest lipid content of 63.4% for M. isabellina and 12.8% for C. sorokiniana, followed by acid digestion. The highest lipid content of C. curvatus was achieved at 47.3% through acid digestion, compared to sonication, bead-beating and Soxhlet methods. This study suggests that acid digestion may be the most simple and effective method for lipid recovery from the tested species in terms of time and reaction temperature. Moreover, fatty acid profile analysis revealed that lipids extracted by acid digestion were similar to those from untreated microbial cells, indicating a favorable feedstock for use in biodiesel production. Practical applications: An effective cell disruption method is critical for not only enhancing the lipid recovery in a large scale but also reporting the accurate values of lipid contents of microorganisms in a lab scale. Different cell disruption methods have different effects on lipid extraction from different microorganisms. In this work, for the first time, we performed comparative investigations among yeast, fungus, and microalga to reveal the lipid recovery yields by cell disruption methods for microbial lipid extraction from different oleaginous microorganisms and identified their most effective methods for analytical lipid determination. An effective cell disruption method is critical for enhancing lipid recovery and reporting the accurate lipid contents of microorganisms. In this study, the authors performed comparative investigations among yeast, fungus, and microalga to determine the lipid recovery yields from these oleaginous micro organisms using various cell disruption methods. They identified the most effective cell disruption methods for analytical lipid determination in each microorganism. [ABSTRACT FROM AUTHOR]

Patterns of the biosynthesis of major metabolites of the oleaginous yeast Cryptococcus curvatus NRRL Y-1511 were investigated during cultivation on sugar-based media. When lactose or sucrose was employed as substrate under nitrogen-limited conditions, the yeast strain accumulated high quantities of intra-cellular total sugars (ITS) at the beginning of fermentation (up to 68% w/w), with ITS values progressively decreasing to 20%, w/w, at the end of the fermentation. Decrease in ITS content and consumption of extra-cellular lactose led to a subsequent rise in lipid accumulation, reaching 29.8% in dry cell weight at 80 g/L of initial lactose concentration. Lactose was a more favorable substrate for lipid production than sucrose. In nitrogen-excess conditions, ITS were produced in significant quantities despite the continuous presence of nitrogen in the medium. Growth on lactose was not followed by secretion of extra-cellular β-galactosidase. High quantities of extra-cellular invertase were observed during growth on sucrose. The composition of ITS was highly influenced by the sugar used as substrate. Cellular lipids contained mainly palmitic and to lesser extent linoleic and stearic acids. This is the first report in the literature that demonstrates the interplay between the biosynthesis of intra-cellular total sugars and lipid synthesis for oleaginous yeast strains. Practical applications: Cryptococcus curvatus represents a promising candidate for the successful production of microbial lipids and polysaccharides during growth on several types of low-cost sugars utilized as substrates. Thus, C. curvatus could be used to produce a variety of metabolites useful for industrial biotechnology. Interplay between the biosynthesis of intra-cellular total sugars produced per unit of dry weight (YITS/X %, w/w) and intra-cellular lipids produced per unit of dry weight (YL/X %, w/w) of Cryptococcus curvatus, during growth on lactose, under nitrogen-limited conditions. [ABSTRACT FROM AUTHOR]

Simultaneous utilization of mixed sugars is one of the major challenges for biofuel production utilizing lignocellulosic biomass as feedstock. Our previous work proved that the oleaginous yeast Cryptococcus curvatus could efficiently produce lipids, the precursors of hydrocarbons with high energy density, from lignocellulosic hydrolysates. However, the strain's capability of simultaneously utilizing primary sugars was still unknown. Thus, this work comprehensively explored the co-utilization of glucose, xylose and cellobiose by C. curvatus . The results indicated that the consumption of both xylose and cellobiose was repressed by glucose, while xylose and cellobiose could be simultaneously consumed at similar rates. The total sugar consumption rates remained constant at about 0.6 g L −1 h −1 regardless of the sugar composition in the mixtures, and the cell biomass and lipid production by C. curvatus cultured on the different sugar mixtures were similar. Moreover, compared with glucose and xylose, cellobiose could lead to higher dry cell weights and lipid yields, suggesting an excellent carbon source for lipid production. Based on these findings, this study demonstrated an effective approach for alleviating glucose repression for microbial lipid production by C. curvatus through xylose/cellobiose co-utilization which would greatly contribute to a more efficient and economical cellulosic biofuel production. [ABSTRACT FROM AUTHOR]

Background Microbial lipids produced from lignocellulosic biomass hold great promise for the biodiesel industry. These lipids usually consist of three major processes: pretreatment, enzymatic hydrolysis and lipid production. However, the conventional strategy of using biomass hydrolysates as the feedstock for lipid production suffers from low lipid coefficient and prohibitively high costs. More cost-effective and integrated processes are required to advance lignocellulosic biomass-based microbial lipid technology. Results Three different strategies were tested using the oleaginous yeast Cryptococcus curvatus ATCC 20509 as a lipid producer and alkaline-pretreated corn stover as a model material. It was found that the separate hydrolysis and enhanced lipid production process required more cellulolytic enzymes yet afforded a low lipid coefficient of 115.6 mg/g pretreated corn stover. When biomass hydrolysis and lipid production were integrated, the amounts of cellulase and xylanase were reduced and no β-glucosidase was required. The simultaneous saccharification and lipid production process gave a lipid coefficient of 129.4 mg/g pretreated corn stover. A higher lipid coefficient of 159.4 mg/g pretreated corn stover was obtained using the simultaneous saccharification and enhanced lipid production (SSELP) process. Furthermore, cellulolytic enzymes were found recoverable and reusable upon recycling the spent supernatants of the SSELP process, which could reduce enzyme consumption and wastewater discharge. Conclusions The SSELP process was superior to other processes in terms of converting alkaline-pretreated corn stover into lipids by C. curvatus, as it required less cellulolytic enzymes and had a higher lipid coefficient. Moreover, the process facilitated easy enzyme recycling that should lead to further reduction of enzyme consumption. These results provide valuable information for cost-effective lipid production from lignocelluloses, which should be particularly important in achieving a sustainable production of biodiesel. [ABSTRACT FROM AUTHOR]

Abstract: Oleaginous microbes which are surely one sustainable way of producing lipid in absolute need of reducing the substrate cost for the commercialization. In this study, algal biomass residue, which is a waste generated after the production of algae-based biofuel, was investigated for its possibility as a cheap feedstock using our newly isolated yeast Cryptococcus curvatus. To make the algae biomass bioavailable, hydrodynamic cavitation (HC) was employed as a pretreatment means, whose optimal treatment condition was systematically set up using Response Surface Methodology (RSM). The optimal condition was HC pretreatment time of 6.88 min at pH 9.47. When used the pretreated microalgae, the C. curvatus can grow with growth rate of 0.584/h, biomass reached to 7.47 g/L in a day, and lipid productivity (1.5 g/L/d) was better than defined medium using commercial chemicals. [Copyright &y& Elsevier]

Highlights: [•] Direct transesterfication of wet yeast cells using methanol and microwave irradiation is feasible. [•] Methanol to biomass ratio, stirring speed and KOH concentration were critical to biodiesel yield. [•] Under optimal conditions, the crude biodiesel contained 64% of FAMEs and was 92% of yeast lipids. [ABSTRACT FROM AUTHOR]

Highlights: [ • ] Study the effect of lignocellulose degradation products on Cryptococcus curvatus. [•] Furfural had the strongest inhibitory effect than any other tested compounds. [•] C. curvatus had its excellent performance on the inhibitor tolerance. [•] 10% (v/v) inoculum size was optimal for C. curvatus in the presence of inhibitors. [•] C. curvatus could utilize the primary sugars derived from lignocellulosic biomass. [Copyright &y& Elsevier]

Abstract: Waste spent yeast from brewery industry was used as a sole growth substrate to grow an oleaginous yeast Cryptococcus curvatus for the purpose of biodiesel production. Approximately 7g/l/d of biomass productivity was obtained using only spent yeast (30g/l) without additional nutrients and pretreatment of any kind. To make best use of available nutrients in the spent yeast, stepwise cultivation was carried out in a batch culture mode and the highest biomass and lipid content, which were 50.4g/l and 37.7%, respectively, were obtained at 35:1 of C/N ratio. Lipid from C. curvatus was found to be a quality-sufficient source of oil as a transportation fuel in terms of cetane, iodine values, and oxidation stability, although the values of cold filter plugging point were less desirable. Economic evaluation revealed that the use of the spent yeast could significantly reduce the unit cost of yeast-based biodiesel production. [Copyright &y& Elsevier]

Abstract: Oleaginous yeast Cryptococcus curvatus has become some of the most promising feedstock for biodiesel production due to their high production efficiency. However, high cost of cultivation, especially substrate cost, hinders rapid commercialization of yeast-based biodiesel. In this study, waste activated sludge (WAS), which is rich in nutrients and organic matters, was examined as an economic substitute for organic substances. To be efficiently bioavailable, WAS must be pretreated. Hydrodynamic cavitation reaction time and pH were identified as experimental factors, whereas growth rate was selected as response parameter. The experimental factors were optimized employing Response Surface Methodology (RSM). Growth rate of 1.13/h was optimized at reaction time 12.5min and pH 8.65. When sludge pretreated under these optimal conditions was used as a substitute to yeast extract, 9.84g/L of biomass was obtained in a day and lipid was accumulated up to 23% of dry weight. [Copyright &y& Elsevier]

Background: Microbial lipid production by using lignocellulosic biomass as the feedstock holds a great promise for biodiesel production and biorefinery. This usually involves hydrolysis of biomass into sugar-rich hydrolysates, which are then used by oleaginous microorganisms as the carbon and energy sources to produce lipids. However, the costs of microbial lipids remain prohibitively high for commercialization. More efficient and integrated processes are pivotal for better techno-economics of microbial lipid technology. Results: Here we describe the simultaneous saccharification and enhanced lipid production (SSELP) process that is highly advantageous in terms of converting cellulosic materials into lipids, as it integrates cellulose biomass hydrolysis and lipid biosynthesis. Specifically, Cryptococcus curvatus cells prepared in a nutrient-rich medium were inoculated at high dosage for lipid production in biomass suspension in the presence of hydrolytic enzymes without auxiliary nutrients. When cellulose was loaded at 32.3 g/L, cellulose conversion, cell mass, lipid content and lipid coefficient reached 98.5%, 12.4 g/L, 59.9% and 204 mg/g, respectively. Lipid yields of the SSELP process were higher than those obtained by using the conventional process where cellulose was hydrolyzed separately. When ionic liquid pretreated corn stover was used, both cellulose and hemicellulose were consumed simultaneously. No xylose was accumulated over time, indicating that glucose effect was circumvented. The lipid yield reached 112 mg/g regenerated corn stover. This process could be performed without sterilization because of the absence of auxiliary nutrients for bacterial contamination. Conclusions: The SSELP process facilitates direct conversion of both cellulose and hemicellulose of lignocellulosic materials into microbial lipids. It greatly reduces time and capital costs while improves lipid coefficient. Optimization of the SSELP process at different levels should further improve the efficiency of microbial lipid technology, which in turn, promote the biotechnological production of fatty acid-derived products from lignocellulosic biomass. [ABSTRACT FROM AUTHOR]

Abstract: Cryptococcus curvatus is able to grow on crude glycerol derived from the biodiesel production process. Through Box–Behnken design and response surface methodology, the optimal temperature, pH, and glycerol concentration for yeast growth and lipid production on pretreated crude glycerol was identified as 30.2 °C, 6.0, and 19.8 g/l, respectively. Adopting these optimal parameters, the biomass density obtained was 7.11 ± 0.36 g/l with a lipid content of 38.53 ± 1.88%, which matched well with predicted values of 6.98 g/l and 41.31%, respectively. The resulting parameters of the response surface method optimization were used in a fed-batch fermentation where crude glycerol was automatically pumped in responding to exhausted oxygen levels in the fermentor. At the end of 12 days, the biomass concentration was 44.53 g/l and the lipid content was 49.0%. Compared with our previous fed-batch experiment which was conducted under un-optimized condition and manual feeding, the yield of biomass and lipid increased 35.26% and 25.29%, respectively. The optimal parameters and the automatic pumping scheme developed from this study will assist greatly in future industrial scale application of producing microbial oils from crude glycerol. [Copyright &y& Elsevier]

Abstract: Cryptococcus curvatus has great potential in fermenting unconditioned hydrolysates of sweet sorghum bagasse. With hydrolysates obtained by enzymatic hydrolysis of the solid pretreated by microwave with lime, the maximal yeast cell dry weight and lipid content were 10.83 g/l and 73.26%, respectively. For hydrolysates obtained in the same way but without lime, these two parameters were 15.50 g/l and 63.98%, respectively. During yeast fermentation, glucose and xylose were consumed simultaneously while cellobiose was released from the residual bagasse. The presence of lime, on one hand, made cellulose more accessible to enzymes as evidenced by higher total reducing sugar release compared to that without during enzymatic hydrolysis step; on the other hand, it caused the degradation of sugars to non-sugar chemicals during pretreatment step. As a result, higher lipid yield of 0.11 g/g bagasse or 0.65 ton/hectare of land was achieved from the pathway of microwave pretreatment and enzymatic hydrolysis while 0.09 g/g bagasse or 0.51 ton/hectare of land was attained from the process of lime-assisted microwave pretreatment followed by the same enzymatic saccharification. [Copyright &y& Elsevier]

Abstract: Culturing oleaginous yeast with organic residues can provide cheap feedstock for biodiesel production, but the high content of nitrogen in some feedstocks (such as food residues) can lead to inhibition to cell growth and lipid synthesis, and this may be caused by ammonia. Thus, the inhibitory effect of ammonia on the growth of yeast Cryptococcus curvatus and its possible mechanism were evaluated in this study. The biomass production, lipid contents, affected metabolic pathways, and activities of the related pathway enzymes were investigated. The results showed that the yeast biomass production on acetate decreased from 5.9 kg m−3 to 2.0 kg m−3 when the initial ammonia concentration (in nitrogen equivalent terms) increased from 131 g m−3 to 3140 g m−3. Ammonia significantly inhibited the yeast growth with volatile fatty acids (VFA) as carbon sources, but not in the culture with pyruvate generating substances (PGS) as carbon sources. However, the fatty acid synthesis was suppressed by ammonia in cultures with either VFA or PGS. The study on mechanism of ammonia inhibition suggested it might include an inhibition on the acyl-CoA synthetase activity, since a 61% decrease in the enzyme activity was found when 3140 g m−3 of ammonia (in nitrogen equivalent terms) was added. This study firstly reported the effect of ammonia with high concentrations to oleaginous yeast growth and lipid accumulation, and provided preliminary evidences on its possible mechanism of acyl-CoA synthetase inhibition. [Copyright &y& Elsevier]

Abstract: The objective of the current report is process optimization for economical production of lipids by the well known oleaginous yeast Cryptococcus curvatus and conversion of the lipids to biodiesel. A high cell density fed-batch cultivation on low cost substrate viz. crude glycerol resulted in a dry biomass and oil yield of up to 69g/L and 48% (w/w), respectively. The process was scaled up easily to 26L. The oil extraction process was also optimized using environmentally safe solvents. The oil profile indicated a high oleic acid content followed by palmitic acid, stearic acid and linoleic acid. The oil was trans-esterified to biodiesel and thoroughly characterized. This is the first end to end report on production of biodiesel from the C. curvatus oil. [Copyright &y& Elsevier]

Abstract: Volatile fatty acids (VFA) from dark fermentation hydrogen production were tested as carbon sources for the culture of oleaginous yeast Cryptococcus curvatus, which is a promising feedstock for biofuel production. The optimal acetate concentration and pH were investigated when potassium acetate was used as the sole carbon source. Comparisons were then made when hydrogen production effluent (HPE) from synthetic wastewater was tested as feedstock. A pH-stat culture fed with acetic acid ultimately produced 168 g/L biomass, with a lipid content of 75.0%. No inhibitor to yeast growth was produced in the hydrogen production process. However, inhibition occurred in culture with HPE from food waste (FW), indicating that inhibitors may be present in the original raw food waste. This inhibition could be avoided by a process that uses glucose as the initial carbon source and then is continuously fed with FW-HPE. The biomass productivity in this continuous culture process reached 0.34 g/L/h, but the lipid content was only 13.5%. These results suggest that FW-HPE alone is not an optimal feedstock, but HPE derived from nitrogen-deficient waste streams could be good feedstocks. This study provides preliminary evidence for the feasibility of using organic waste for the co-production of hydrogen and lipid. [Copyright &y& Elsevier]

Abstract: This paper explores the use of the hydrolysate from the dilute sulfuric acid pretreatment of wheat straw for microbial oil production. The resulting hydrolysate was composed of pentoses (24.3g/L) and hexoses (4.9g/L), along with some other degradation products, such as acetic acid, furfural, and hydroxymethylfurfural (HMF). Five oleaginous yeast strains, Cryptococcus curvatus, Rhodotorula glutinis, Rhodosporidium toruloides, Lipomyces starkeyi, and Yarrowia lipolytica, were evaluated by using this hydrolysate as substrates. The results showed that all of these strains could use the detoxified hydrolysate to produce lipids while except R. toruloides non-detoxified hydrolysate could also be used for the growth of all of the selective yeast strains. C. curvatus showed the highest lipid concentrations in medium on both the detoxified (4.2g/L) and non-detoxified (5.8g/L) hydrolysates. And the inhibitory effect studies on C. curvatus indicated HMF had insignificant impacts at a concentration of up to 3g/L while furfural inhibited cell growth and lipid content by 72.0% and 62.0% at 1g/L, respectively. Our work demonstrates that lipid production is a promising alternative to utilize hemicellulosic sugars obtained during pretreatment of lignocellulosic materials. [Copyright &y& Elsevier]

Abstract: Cryptococcus curvatus, an oleaginous yeast was observed to grow on crude glycerol derived from yellow grease. When cultured in a one-stage fed-batch process wherein crude glycerol and nitrogen source were fed intermittently for 12days, the final biomass density and lipid content were 31.2g/l and 44.2%, respectively. When cultured in a two-stage fed-batch operation wherein crude glycerol was supplemented at different time points while nitrogen source addition was discontinued at the middle of the experiment, the biomass density was 32.9g/l and the lipid content was 52% at the end of 12days. Compared with other oil feedstocks for biodiesel production, lipid accumulated by C. curvatus grown on glycerol has high concentration of monounsaturated fatty acid, which makes it an excellent source for biodiesel use. [Copyright &y& Elsevier]

• Significant lipid and total dry biomass production were recorded by a novel Cryptococcus curvatus strain growing on commercial xylose. • Single cell oil produced was an appropriate material for the synthesis of high-quality biodiesel. • Lipid production bioprocess was successfully simulated by numerical models. • Biomass and laccase production of edible and medicinal mushrooms was achieved using lipid fermentation waste-waters. A rarely studied Cryptoccus curvatus strain (viz. NRRL Y-1511) was cultivated in shake-flask submerged nitrogen-limited experiments, with commercial xylose being employed as substrate and has been proved as quite appropriate for total biomass, lipid and endopolysaccharide production. A batch bioreactor trial compared to the equivalent shake-flask experiment showed larger amounts of total lipid and biomass produced. Total dry biomass of 26.1 g/L containing c. 30 % of lipids was synthesized. Endopolysaccharides in relatively high concentrations were synthesized in the early growth stages decreasing afterwards in favor of storage lipid synthesis. Lipid production bioprocess was successfully simulated by a modified Velruyten-Aggelis model and the optimized parameter values were found to be similar to those experimentally measured. Lipids contained mostly neutral fractions (triacylglycerols) and were rich in the fatty acid Δ9C18:1, constituting a suitable starting material for the synthesis of high-quality biodiesel. Within the frame of bio-refinery, lipid fermentation waste-waters were used as maceration waters for a novel mushroom cultivation process of Pleurotus ostreatus , Lentinula edodes and Ganoderma adspersum edible and medicinal mushrooms. These waste-waters were used for first time in mushroom cultivation process demonstrating positive effect on laccase and sporophore production by these mushroom species. [ABSTRACT FROM AUTHOR]