Search

Background Microbial lipids derived from various lignocellulosic feedstocks have emerged as a promising candidate for the biodiesel industry and a potential substitute for high value-added fats. However, lignocellulosic biomass, especially herbaceous biomass, such as water hyacinth, contains high concentrations of nitrogenous components. These compounds impede microbial lipid production, as lipid biosynthesis is commonly induced by imposing a nutrient deficiency, especially nitrogen starvation. Novel strategies and bioprocesses are pivotal for promoting lipid production from nitrogen-rich biomass. Results Here a combined strategy of phosphate removal and acetate supplementation was described for enhanced microbial lipid production on water hyacinth hydrolysates by Cutaneotrichosporon oleaginosum (formerly Cryptococcus curvatus). Lipid production was significantly improved, when the phosphorus limitation and sugars/acetate co-utilization strategies were used separately. In this case, acetate and glucose were consumed simultaneously. Lipid production was observed by the combination of phosphate removal with acetate supplementation. Lipid titer, content, and yield were determined to be 7.3 g/L, 59.7% and 10.1 g/100 g raw water hyacinth, respectively. These data were increased by 4.2, 4.6, and 4.3 times, respectively, compared to those from the unprocessed hydrolysates. The fatty acid compositions of the resulting lipids bear a marked resemblance to those of rapeseed oil, indicating their applicability to the biodiesel industry. Conclusions The combination of phosphate removal and acetate supplementation was successful in significantly enhancing microbial lipid production. This strategy offers a valuable solution for nitrogen-rich lignocellulosic feedstocks utilization, which should foster more economical nitrogen-rich biomass-to-lipid bioprocesses.

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.

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.

Background: Oleaginous microorganisms, such as different yeast and algal species, can represent a sustainable alternative to plant oil for the production of biodiesel. They can accumulate fatty acids (FA) up to 70% of their dry weight with a predominance of (mono)unsaturated species, similarly to what plants do, but differently from animals. In addition, their growth is not in competition either with food, feed crops, or with agricultural land. Despite these advantages, the exploitation of the single cell oil system is still at an early developmental stage. Cultivation mode and conditions, as well as lipid extraction technologies, represent the main limitations. The monitoring of lipid accumulation in oleaginous microorganisms is consequently crucial to develop and validate new approaches, but at present the majority of the available techniques is time consuming, invasive and, when relying on lipid extraction, can be affected by FA degradation. Results: In this work the fatty acid accumulation of the oleaginous yeasts Cryptococcus curvatus and Rhodosporidium toruloides and of the non-oleaginous yeast Saccharomyces cerevisiae (as a negative control) was monitored in situ by Fourier Transform Infrared Spectroscopy (FTIR). Indeed, this spectroscopic tool can provide complementary information to those obtained by classical techniques, such as microscopy, flow cytometry and gas chromatography. As shown in this work, through the analysis of the absorption spectra of intact oleaginous microorganisms it is possible not only to monitor the progression of FA accumulation but also to identify the most represented classes of the produced lipids. Conclusions: Here we propose FTIR microspectroscopy - supported by multivariate analysis - as a fast, reliable and non invasive method to monitor and analyze FA accumulation in intact oleaginous yeasts. The results obtained by the FTIR approach were in agreement with those obtained by the other classical methods like flow cytometry and gas chromatography. Moreover, the possibility to track lipid production in real time is highly desirable to support the initial screening of strains and media as well as to optimize the scaling up experiments, which are essential for a viable and successful development of an industrial production process. © 2014 Ami et al.; licensee BioMed Central Ltd.

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.

Background Oleaginous microorganisms are attractive feedstock for production of liquid biofuels. Direct hydrothermal liquefaction (HTL) is an efficient route that converts whole, wet biomass into an energy-dense liquid fuel precursor, called ‘biocrude’. HTL represents a promising alternative to conventional lipid extraction methods as it does not require a dry feedstock or additional steps for lipid extraction. However, high operating pressure in HTL can pose challenges in reactor sizing and overall operating costs. Through the use of co-solvents the HTL operating pressure can be reduced. The present study investigates low-temperature co-solvent HTL of oleaginous yeast, Cryptococcus curvatus, using laboratory batch reactors. Results In this study, we report the co-solvent HTL of microbial yeast biomass in an isopropanol–water binary system in the presence or absence of Na2CO3 catalyst. This novel approach proved to be effective and resulted in significantly higher yield of biocrude (56.4 ± 0.1 %) than that of HTL performed without a co-solvent (49.1 ± 0.4 %)(p = 0.001). Addition of Na2CO3 as a catalyst marginally improved the biocrude yield. The energy content of the resulting biocrude (~37 MJ kg−1) was only slightly lower than that of petroleum crude (42 MJ kg−1). The HTL process was successful in removing carboxyl groups from fatty acids and creating their associated straight-chain alkanes (C17–C21). Experimental results were leveraged to inform techno-economic analysis (TEA) of the baseline HTL conversion pathway to evaluate the commercial feasibility of this process. TEA results showed a renewable diesel fuel price of $5.09 per gallon, with the HTL-processing step accounting for approximately 23 % of the total cost for the baseline pathway. Conclusions This study shows the feasibility of co-solvent HTL of oleaginous yeast biomass in producing an energy-dense biocrude, and hence provides a platform for adding value to the current dairy industry. Co-solvents can be used to lower the HTL temperature and hence the operating pressure. This process results in a higher biocrude yield at a lower HTL temperature. A conceptual yeast HTL biofuel platform suggests the use of a dairy waste stream for increasing the productivity and sustainability of rural areas while providing a new feedstock (yeast) for generating biofuels. Electronic supplementary material The online version of this article (doi:10.1186/s13068-015-0345-5) contains supplementary material, which is available to authorized users.

Background The feasibility of heterotrophic–phototrophic symbioses was tested via pairing of yeast strains Cryptococcus curvatus, Rhodotorula glutinis, or Saccharomyces cerevisiae with a sucrose-secreting cyanobacterium Synechococcus elongatus. Results The phototroph S. elongatus showed no growth in standard BG-11 medium with yeast extract, but grew well in BG-11 medium alone or supplemented with yeast nitrogen base without amino acids (YNB w/o aa). Among three yeast species, C. curvatus and R. glutinis adapted well to the BG-11 medium supplemented with YNB w/o aa, sucrose, and various concentrations of NaCl needed to maintain sucrose secretion from S. elongatus, while growth of S. cerevisiae was highly dependent on sucrose levels. R. glutinis and C. curvatus grew efficiently and utilized sucrose produced by the partner in co-culture. Co-cultures of S. elongatus and R. glutinis were sustained over 1 month in both batch and in semi-continuous culture, with the final biomass and overall lipid yields in the batch co-culture 40 to 60% higher compared to batch mono-cultures of S. elongatus. The co-cultures showed enhanced levels of palmitoleic and linoleic acids. Furthermore, cyanobacterial growth in co-culture with R. glutinis was significantly superior to axenic growth, as S. elongatus was unable to grow in the absence of the yeast partner when cultivated at lower densities in liquid medium. Accumulated reactive oxygen species was observed to severely inhibit axenic growth of cyanobacteria, which was efficiently alleviated through catalase supply and even more effectively with co-cultures of R. glutinis. Conclusions The pairing of a cyanobacterium and eukaryotic heterotroph in the artificial lichen of this study demonstrates the importance of mutual interactions between phototrophs and heterotrophs, e.g., phototrophs provide a carbon source to heterotrophs, and heterotrophs assist phototrophic growth and survival by removing/eliminating oxidative stress. Our results establish a potential stable production platform that combines the metabolic capability of photoautotrophs to capture inorganic carbon with the channeling of the resulting organic carbon directly to a robust heterotroph partner for producing biofuel and other chemical precursors. Electronic supplementary material The online version of this article (doi:10.1186/s13068-017-0736-x) contains supplementary material, which is available to authorized users.

Background Oligocelluloses and oligoxyloses are partially hydrolyzed products from lignocellulosic biomass hydrolysis. Biomass hydrolysates usually contain monosaccharides as well as various amounts of oligosaccharides. To utilize biomass hydrolysates more efficiently, it is important to identify microorganisms capable of converting biomass-derived oligosaccharides into biofuels or biochemicals. Results We have demonstrated that the oleaginous yeast Cryptococcus curvatus can utilize either oligocelluloses or oligoxyloses as sole carbon sources for microbial lipid production. When oligocelluloses were used, lipid content and lipid coefficient were 35.9% and 0.20 g/g consumed sugar, respectively. When oligoxyloses were used, lipid coefficient was 0.17 g/g consumed sugar. Ion chromatography analysis showed oligocelluloses with a degree of polymerization from 2 to 9 were assimilated. Our data suggested that these oligosaccharides were transported into cells and then hydrolyzed by cytoplasmic enzymes. Further analysis indicated that these enzymes were inducible by oligocelluloses. Lipid production on cellulose by C. curvatus using the simultaneous saccharification and lipid production process in the absence of cellobiase achieved essentially identical results to that in the presence of cellobiase, suggesting that oligocelluloses generated in situ were utilized with high efficiency. This study has provided inspiring information for oligosaccharides utilization, which should facilitate biorefinery based on lignocellulosic biomass. Conclusions C. curvatus can directly utilize biomass-derived oligosaccharides. Oligocelluloses are transported into the cells and then hydrolyzed by cytoplasmic enzymes. A simultaneous saccharification and lipid production process can be conducted without oligocelluloses accumulation in the absence of cellobiase by C. curvatus, which could reduce the enzyme costs.