Cost-effective optimized scleroglucan production by Sclerotium rolfsii ATCC 201126 at bioreactor scale. A quantity-quality assessment.

Exopolysaccharide (EPS) secretion by Sclerotium rolfsii ATCC 201126 in submerged cultures, already identified as high-osmolarity responsive, was assessed by reducing C-source without compromising EPS yields. A designed medium with 80 g sucrose L-1 (MOPT80) was tested at 3 L-bioreactor scale at different temperature, agitation, aeration and pH (uncontrolled vs. controlled) values. Optimal operative conditions (200 rpm, 28 °C, 0.5 vvm and initial pH -pHi- 4.5) were validated, as well as the possibility to work at pHi 5.5 to reduce biomass production. Purified EPSs produced in MOPT80 at optimal and other valid operative conditions exhibited refined grade (<1 % proteins and ash, 3-4 % reducing sugars, 87-99 % total sugars). EPS purity, MW and rheological parameters led to discourage pH controlled at 4.5. Relatively constant MW (6-8 × 106 Da) and outstanding viscosifying ability were found. Polyphasic EPS analysis (titre, purity, macromolecular features and rheological fitness) would support to properly select production conditions.

Single Cell Oil Production from Undetoxified Arundo donax L. hydrolysate by Cutaneotrichosporon curvatus.

The use of low-cost substrates represents one key issue to make single cell oil production sustainable. Among low-input crops, Arundo donax L. is a perennial herbaceous rhizomatous grass containing both C5 and C6 carbohydrates. The scope of the present work was to investigate and optimize the production of lipids by the oleaginous yeast Cutaneotrichosporon curvatus from undetoxified lignocellulosic hydrolysates of steam-pretreated A. donax. The growth of C. curvatus was first optimized in synthetic media, similar in terms of sugar concentration to hydrolysates, by applying the response surface methodology (RSM) analysis. Then the bioconversion of undetoxified hydrolysates was investigated. A fed-batch process for the fermentation of A. donax hydrolysates was finally implemented in a 2-L bioreactor. Under optimized conditions, the total lipid content was 64% of the dry cell weight and the lipid yield was 63% of the theoretical. The fatty acid profile of C. curvatus triglycerides contained 27% palmitic acid, 33% oleic acid and 32% linoleic acid. These results proved the potential of lipid production from A. donax, which is particularly important for their consideration as substitutes for vegetable oils in many applications such as biodiesel or bioplastics.

Assessment of white rot fungus mediated hardwood degradation by FTIR spectroscopy and multivariate analysis.

Evaluating the biomass degradation using fast, validate and sensitive techniques for exploratory purposes of biofuel production has been developed since last decade. Thus, we assessed the degradation of two Indian hardwoods using FTIR and chemometric approaches. Two white rot fungi, namely Pseudolagarobasidium acaciicola AGST3 and Tricholoma giganteum AGDR1, were selected among twenty-one fungal isolates for higher hardwood degradation. In the screening, P. acaciicola AGST3 and T. giganteum AGDR1 depicted the dry woody mass loss of 20.51% and 22.38%, respectively. Cellulose crystallinity of P. acaciicola AGST3 treated hardwoods was 4-fold lower than untreated hardwoods, showing the higher cellulose degradation efficiency. P. acaciicola AGST3 treated samples exhibited maximum deviation of guaiacyl units of lignin, cellulose and hemicelluloses. T. giganteum AGDR1 treated hardwoods showed maximum deviation of guaiacyl- and syringyl- units of lignin and hemicelluloses. Multivariate approach revealed the degradation patterns and preferences are varied based on the fungi and hardwood. The approach used in the present study can certainly distinguish the variations among the different biomass samples that having similar composition. Additionally, higher lignin degradability of these fungi can be used in biomass pretreatment, which significantly utilized to produce second-generation biofuels.

Biopolymer from marine Athelia and its application on heavy oil recovery in heterogeneous reservoir.

Biopolymer produced from marine Athelia strain presented unique Pseudoplastic behaviors under extremely-high temperature and salinity conditions. Characteristic analysis with FT-IR spectroscopy, high performance liquid chromatography, 1H and 13C NMR and two-dimensional COSY and HMQC spectra showed the structure of ß-(1-6) glucans. Single-factor and orthogonal experiment design were used to optimize the yield, the maximum yield of the biopolymer was 28.32 g/L with 56.64% carbon conversion rate under optimized conditions. Economic investigation demonstrated that this novel biopolymer has great potential of commercialization with the competitive cost of $2896.04-5228.94 per ton for powder. Resistance factor and residual resistance factor were evaluated with core flooding experiments showed that this biopolymer had excellent performance of plugging capacity and profile modification, and indicating the great potential of application on heavy oil recovery.

Study on Lipid Accumulation in Novel Oleaginous Yeast Naganishia liquefaciens NITTS2 Utilizing Pre-digested Municipal Waste Activated Sludge: a Low-cost Feedstock for Biodiesel Production.

The economical production of lipids is considered as an appropriate renewable alternative feedstock for biodiesel production because of the contemporary concerns on fuel crisis, climate change and food security. In this study, lipid accumulation potential of a novel oleaginous yeast isolate Naganishia liquefaciens NITTS2 by utilizing pre-digested municipal waste activated sludge (PWAS) was explored. Optimization of culture conditions was performed using response surface methodology coupled with genetic algorithm and maximum lipid content of 55.7% was obtained. The presence of lipid was visually confirmed by fluorescence microscopy and its characteristic profile was determined by GC-MS. The yeast lipid was recovered and converted into biodiesel by garbage lipase with the efficiency of 88.34 ± 1.2%, which was further analyzed by proton nuclear magnetic resonance spectroscopy. Hence, the results of this study strongly suggest the possibility of using PWAS as an efficient and low-cost resource for the production of biodiesel from the oleaginous yeast.

Exploitation of genus Rhodosporidium for microbial lipid production.

Oleaginous microorganisms are receiving significant attention worldwide for their utility in biodiesel production and the potentiality to produce some specialty-type lipids. There is an increasing interest in isolation/adaption of robust microbe strains and design of innovative fermentation processes to make microbial lipid production a more efficient and economically feasible bio-process. Currently, the genus Rhodosporidium has been considered an important candidate, for the reason that several strains belonging to this genus have shown excellent capabilities of lipid accumulation, broad adaptabilities to various substrates, and co-production of some carotenoids. This paper reviews the current trends in the exploitation of Rhodosporidium species for microbial lipid production, including the utilization of various (single or mixed, pure or waste-derived) substrates, progress of genetic modification and metabolic engineering, innovations in fermentation mode, lipid characterizations and their potential applications. Finally, the constraints and perspectives of cultivating Rhodosporidium species for lipid production are also discussed.

Effect of different lignocellulosic wastes on Hericium americanum yield and nutritional characteristics.

BACKGROUND: The aim of this study was to investigate the possibility of using cottonseed hulls (CSH) and olive press cake (OPC) as new supplement materials for substrate preparation in Hericium americanum cultivation. Some chemical properties of the substrates prepared by mixtures of oak sawdust (OS) with wheat bran (WB), CSH and OPC in different ratios were determined. In addition, the effect of mixtures of OS:CSH and OS:OPC on spawn run time, yield and biological efficiency (BE), average mushroom weight and nutrition content of the fruiting body were compared with the control substrate (8OS:2WB). RESULTS: The yield, BE and average mushroom weight of substrates containing CSH and OPC were higher than the control substrate and increased with an increase in the rate of CSH and OPC in the mixtures. Hericium americanum showed (on a dry weight basis) 8.5-23.7% protein, 9.9-21.2 g kg-1 P, 26.6-35.8 g kg-1 K, 0.63 - 1.33 g kg-1 Mg, 0.19 - 0.23 g kg-1 Ca, 1.34-1.78 g kg-1 Na, 49.5-72.2 mg kg-1 Fe, 6.22-10.11 mg kg-1 Mn, 32.8-82.8 mg kg-1 Zn and 8.6-11.2 mg kg-1 Cu on different growing substrates. The nutritional value of mushrooms was greatly affected by the growing media. CONCLUSION: The results revealed that CSH and OPC could be used as new supplement materials for substrate preparation in H. americanum cultivation. © 2016 Society of Chemical Industry.

Mussel processing wastewater: a low-cost substrate for the production of astaxanthin by Xanthophyllomyces dendrorhous.

BACKGROUND: The use of astaxanthin in different industries such as the chemical, pharmaceutical, food, animal feed and cosmetic has been receiving increasing attention in recent years. Natural supplies of the pigment include crustacean by-products, algal, and microbial cultivation, being the yeast Xanthophyllomyces dendrorhous together with the alga Haematococcus pluvialis the most promising microorganisms for this bioproduction. Different vegetable by-products of the food industry have been explored so far as low-cost substrates for the production of astaxanthin by X. dendrorhous. This study focuses for the first time on the use of a low-cost formulated medium from a marine by-product, mussel-processing wastewater, for the production of astaxanthin by the yeast X. dendrorhous. RESULTS: The yeast was able to grow in non-saccharified mussel broth, revealing the ability of the microorganism to hydrolyze glycogen. However, partial glycogen saccharification with α-amylase was needed for astaxanthin biosynthesis, obtaining maximal productions of 22.5-26.0 mg/L towards the end of the culture and coinciding with yeast highest amylolytic activity. Cultivations in totally-saccharified media revealed an increase in maximal cell concentrations and a decrease in maximal growth rates and astaxanthin production with increasing glucose initial concentration. CONCLUSIONS: Astaxanthin production was higher in partially-saccharified mussel-processing waste than in synthetic medium (yeast peptone dextrose) containing glucose as carbon source (13 mg/L), suggesting this by-product is a promising nutritive medium for astaxanthin production. The use of this effluent also contributes towards the recycling and depuration of this highly pollutant effluent.

Macaúba (Acrocomia aculeata) cake from biodiesel processing: a low-cost substrate to produce lipases from Moniliella spathulata R25L270 with potential application in the oleochemical industry.

BACKGROUND: Biodiesel industry wastes were evaluated as supplements for lipase production by Moniliella spathulata R25L270, which is newly identified yeast with great lipolytic potential. Macaúba cake (MC), used for the first time in this work as inducer to produce lipases, and residual oil (RO) were mixed to maximise enzyme production. The lipase secreted was biochemically characterised. RESULTS: The best ratio for the mixture (MC:RO) was 0.66:0.34 and the fitted values for lipase activity and total protein concentration were 0.98 U mL(-1) and 0.356 mg mL(-1), respectively. Maximum activity obtained (2.47 U mL(-1)) was achieved at 31.5°C and pH 6.7, and the enzyme was stable in this condition. A novel enzyme was purified and identified for the first time by mass spectrometry. The lipase efficiently hydrolysed different natural oils and exhibited selectivity in the production of eicosapentaenoic acid from fish oil. CONCLUSION: The use of MC and RO as a supplement to produce the new lipase from M. spathulata R25L270 may be one alternative for reducing lipase production costs and simultaneously adding value to biodiesel industry residues. The potential application of the lipase in the oleochemical industry was demonstrated by its pH and temperature stabilities and selective hydrolysis.

Selecting low-cost carbon sources for carotenoid and lipid production by the pink yeast Rhodosporidium toruloides NCYC 921 using flow cytometry.

The present work studied low-cost carbon sources for carotenoid and lipid production using the yeast Rhodosporidum toruloides NCYC 921. Carob pulp syrup and sugarcane molasses at different concentrations were used as low-cost carbon sources in R. toruloides batch cultivations. Carob pulp syrup containing a total sugar concentration of 75 g L(-1) induced the highest total fatty acid productivity (1.90 g L(-1)h(-1)) and the highest carotenoid productivity (9.79 µg L(-1)h(-1)). Flow cytometric analysis revealed that most of the yeast cells (>60%) grown on carob pulp syrup displayed intact polarised membranes, conversely to the cells grown on sugarcane molasses, wherein a large proportion (>45%) displayed permeabilised cytoplasmic membranes.

Chitosan-guar gum-silver nanoparticles hybrid matrix with immobilized enzymes for fabrication of beta-glucan and glucose sensing photometric flow injection system.

Simple and fast photometric flow injection analysis system was developed for sensing of ß-1,3-glucan from medicinal mushroom Ganoderma lucidum during fermentation. For this purpose, the chitosan-guar gum-silver nanoparticle-beta glucanase (Ch-GG-AgNPs-ßG) beads and Ch-GG-AgNPs-GOD (glucose oxidase) beads were prepared. The bead packed mini-columns were then used to assemble a flow injection analysis (FIA) system for the detection of ß-(1→3)-d-glucan biomarker or glucose. This colorimetric flow system can detect glucose and glucan with detection limits as low as 50ngmL(-1) and 100ngmL(-1) (S/N=3), respectively. The analysis time of this FIA was approximately 40s, which is faster than the previously reported glucan sensors. The glucose and glucan calibration curves were obtained in the range of 0.25-1.25µgmL(-1) (R(2)=0.988) and 0.2-1.0µgmL(-1)(R(2)=0.979), respectively. The applicability of the nano-bio-composite FIA sensor system for spiked and real ß-(1→3)-d-glucan samples were tested, and the accuracy of the results were greater than 95%. Thus, the designed FIA provides a simple, interference free and rapid tool for monitoring glucose and ß-glucan content, which can be used for various food samples with a little modification.

Mercury in bay bolete (Xerocomus badius): bioconcentration by fungus and assessment of element intake by humans eating fruiting bodies.

Concentrations of mercury (Hg) were determined in 221 specimens of bay bolete and in 221 samples of corresponding forest topsoil layer (0-10 cm) collected from 18 sites across geographically and industrially diverse regions of Poland in 2000-2008. Mercury concentrations in caps and stipes of bay bolete were strongly correlated (p < 0.0001), whereas the relationship between the Hg concentrations in soil and mushrooms varied depending on the sampling location. The bay bolete showed a lower bioconcentration potential of Hg at sites with elevated soil Hg concentrations and a higher bioconcentration at sites with lower Hg concentrations in soil. In view of Hg content, the consumption of bay boletes (caps or whole mushrooms) at the regions surveyed at least at a rate up to 2.5 kg per capita weekly in a mushrooming season is safe and will not result in exceeding of currently allowable Hg intake doses.

13C pulse-labeling assessment of the community structure of active fungi in the rhizosphere of a genetically starch-modified potato (Solanum tuberosum) cultivar and its parental isoline.

• The aim of this study was to gain understanding of the carbon flow from the roots of a genetically modified (GM) amylopectin-accumulating potato (Solanum tuberosum) cultivar and its parental isoline to the soil fungal community using stable isotope probing (SIP). • The microbes receiving (13)C from the plant were assessed through RNA/phospholipid fatty acid analysis with stable isotope probing (PLFA-SIP) at three time-points (1, 5 and 12 d after the start of labeling). The communities of Ascomycota, Basidiomycota and Glomeromycota were analysed separately with RT-qPCR and terminal restriction fragment length polymorphism (T-RFLP). • Ascomycetes and glomeromycetes received carbon from the plant as early as 1 and 5 d after labeling, while basidiomycetes were slower in accumulating the labeled carbon. The rate of carbon allocation in the GM variety differed from that in its parental variety, thereby affecting soil fungal communities. • We conclude that both saprotrophic and mycorrhizal fungi rapidly metabolize organic substrates flowing from the root into the rhizosphere, that there are large differences in utilization of root-derived compounds at a lower phylogenetic level within investigated fungal phyla, and that active communities in the rhizosphere differ between the GM plant and its parental cultivar through effects of differential carbon flow from the plant.

Low-cost production of 6G-fructofuranosidase with high value-added astaxanthin by Xanthophyllomyces dendrorhous.

The effects of medium composition and culture conditions on the production of (6)G-fructofuranosidase with value-added astaxanthin were investigated to reduce the capital cost of neo-fructooligosaccharides (neo-FOS) production by Xanthophyllomyces dendrorhous. The sucrose and corn steep liquor (CSL) were found to be the optimal carbon source and nitrogen source, respectively. CSL and initial pH were selected as the critical factors using Plackett-Burman design. Maximum (6)G-fructofuranosidase 242.57 U/mL with 5.23 mg/L value-added astaxanthin was obtained at CSL 52.5 mL/L and pH 7.89 by central composite design. Neo-FOS yield could reach 238.12 g/L under the optimized medium conditions. Cost analysis suggested 66.3% of substrate cost was reduced compared with that before optimization. These results demonstrated that the optimized medium and culture conditions could significantly enhance the production of (6)G-fructofuranosidase with value-added astaxanthin and remarkably decrease the substrate cost, which opened up possibilities to produce neo-FOS industrially.

Effects of twice-ambient carbon dioxide and nitrogen amendment on biomass, nutrient contents and carbon costs of Norway spruce seedlings as influenced by mycorrhization with Piloderma croceum and Tomentellopsis submollis.

Elevated tropospheric CO(2) concentrations may increase plant carbon fixation. In ectomycorrhizal trees, a considerable portion of the synthesized carbohydrates can be used to support the mutualistic fungal root partner which in turn can benefit the tree by increased nutrient supply. In this study, Norway spruce seedlings were inoculated with either Piloderma croceum (medium distance "fringe" exploration type) or Tomentellopsis submollis (medium distance "smooth" exploration type). We studied the impact of either species regarding fungal biomass production, seedling biomass, nutrient status and nutrient use efficiency in rhizotrons under ambient and twice-ambient CO(2) concentrations. A subset was amended with ammonium nitrate to prevent nitrogen imbalances expected under growth promotion by elevated CO(2). The two fungal species exhibited considerably different influences on growth, biomass allocation as well as nutrient uptake of spruce seedlings. P. croceum increased nutrient supply and promoted plant growth more strongly than T. submollis despite considerably higher carbon costs. In contrast, seedlings with T. submollis showed higher nutrient use efficiency, i.e. produced plant biomass per received unit of nutrient, particularly for P, K and Mg, thereby promoting shoot growth and reducing the root/shoot ratio. Under the given low soil nutrient availability, P. croceum proved to be a more favourable fungal partner for seedling development than T. submollis. Additionally, plant internal allocation of nutrients was differently influenced by the two ECM fungal species, particularly evident for P in shoots and for Ca in roots. Despite slightly increased ECM length and biomass production, neither of the two species had increased its capacity of nutrient uptake in proportion to the rise of CO(2). This lead to imbalances in nutritional status with reduced nutrient concentrations, particularly in seedlings with P. croceum. The beneficial effect of P. croceum thus diminished, although the nutrient status of its host plants was still above that of plants with T. submollis. We conclude that the imbalances of nutrient status in response to elevated CO(2) at early stages of plant development are likely to prove particularly severe at nutrient-poor soils as the increased growth of ECM cannot cover the enhanced nutrient demand. Hyphal length and biomass per unit of ectomycorrhizal length as determined for the first time for P. croceum amounted to 6.9 m cm(-1) and 6.0 µg cm(-1), respectively, across all treatments.

A method for the assessment of the quality of mitochondrial preparations.

A new experimental approach was applied to the quantitative assessment of the physiological properties of mitochondria. An analytical function (QUAL) was derived from a linear combination of parameters selected by factor correspondence analysis (FCA). The quality of a mitochondrial preparation can be expressed by two numerical values computed from the function QUAL: a mean value and its variance. These statistical parameters can be used for exploring data from planned experiments.