Volatile profiles of healthy and aflatoxin contaminated pistachios.

The volatile profile of four different groups of dried pistachios namely: H: healthy, NC: naturally contaminated with aflatoxin, AC: artificially contaminated with aflatoxigenic strains of the fungi Aspergillus flavus and ANT: artificially contaminated with non-toxigenic strains of the fungi A. flavus, was determined. The volatiles were isolated by the HS-SPME method and determined by GC-FID and GC-MS, whereas aflatoxin by HPLC. Seventy two volatile compounds were identified almost equally distributed among the above four studied groups. The predominant chemical compounds were monoterpenes, alcohols, ketones, aldehydes, esters and hydrocarbons. The monoterpenes, mainly determined as α-pinene and α-terpinolene were detected in all samples. Even though the general volatile profile was similar among groups, some differences were detected between healthy and contaminated groups of samples. When some key volatiles such as eight-carbon and seven-carbon alcohols and aldehydes were used along with the species-specific sesquiterpenes and the other terpenes detected, a correct classification was obtained in H, NC, AC and ANT groups, as was demonstrated by cluster and discriminant analyses. This evidence provides a potential tool for distinguishing contaminated samples on the basis of characteristic volatile patterns.

Iron or zinc dialyzability obtained from a modified in vitro digestion procedure compare well with iron or zinc absorption from meals.

The dialyzability method has been considered a convenient option for screening a large number of samples for iron bioavailability. A new setup that allows the rapid and efficient application of the dialyzability method has been recently proposed. To evaluate the new setup for its applicability in predicting iron and zinc bioavailability, dialyzable iron or zinc were compared with iron or zinc absorption. A series of solutions and meals that have been employed in published studies on iron or zinc absorption were prepared as described in the literature and digested in vitro using the new setup. Dialyzability was measured and correlated with published data on absorption. Iron or zinc dialyzability measured with the proposed setup correlated well with iron or zinc absorption. These results suggest that the new setup can be employed in the future applications for the prediction of iron or zinc bioavailability.

Organic nitrogen of tomato waste hydrolysate enhances glucose uptake and lipid accumulation in Cunninghamella echinulata.

AIMS: To investigate the effect of organic nitrogen on lipogenesis during growth of Cunninghamella echinulata on tomato waste hydrolysate (TWH) media. METHODS AND RESULTS: Cunninghamella echinulata grown on a TWH medium rapidly took up glucose and produced large amounts of lipids. However, when some quantities of the organic nitrogen were removed from TWH (by acid followed by alkaline precipitation of proteins) the uptake of glucose was dramatically reduced and large quantities of fungal biomass having low lipid content were produced. Nevertheless, when glycerol was used as carbon source instead of glucose, the uptake rate as well as the biomass production and the lipid accumulation processes were unaffected by the TWH organic nitrogen removal. Finally, when the fungus was grown on a glucose supplemented TWH medium that contained no assimilable organic nitrogen (after further precipitation of proteins with methanol), the produced biomass contained non-negligible quantities of lipids, although glucose uptake remained low. Lipid analysis showed that the produced lipids comprised mainly of neutral lipids, which were preferentially consumed during lipid turnover. Lipid production on the original TWH medium having glucose as carbon source was 0.48 g of lipid per gram of dry biomass, corresponding to 8.7 g of lipid per litre of growth medium. The produced lipids contained 11.7%gamma-linolenic acid (GLA), hence the GLA yield was more than 1 g l(-1). CONCLUSIONS: Organic nitrogen compounds found in TWH favour glucose (but not glycerol) uptake and lipid accumulation in C. echinulata. SIGNIFICANCE AND IMPACT OF THE STUDY: Agro-industrial wastes containing organic nitrogen, such as tomato waste, are produced in vast amounts causing severe environmental problems. These wastes could be used as fermentation feedstock to produce microbial lipids.

RP-HPLC analysis of the phenolic compounds of plant extracts. investigation of their antioxidant capacity and antimicrobial activity.

Extracts of aromatic plants of Greek origin were examined as potential sources of phenolic compounds. RP-HPLC with UV detection was employed for the identification and quantification of the phenolic antioxidants, present in methanolic extracts. The most abundant phenolic acids were ferulic acid (1.1-280 mg/100 g of dry sample) and caffeic acid (1.2-60 mg/100 g of dry sample). (+)-Catechin and quercetin were the most abundant flavonoids. Apigenin and luteolin were detected in high amounts in Menta pulegium and Thymus vulgaris, respectively. The antioxidant capacity was determined, in dried ground plants and in their methanol extracts, with the Rancimat test using sunflower oil as substrate. Both pulverized plants and extracts showed antioxidant capacity. Total phenolic content in the extracts was determined spectrometrically according to the Folin-Ciocalteu assay and ranged from 1 to 21 mg of gallic acid/100 g of dry sample. Antimicrobial activity of the extracts against selected microbes was also conducted in this study.

Repression of reserve lipid turnover in Cunninghamella echinulata and Mortierella isabellina cultivated in multiple-limited media.

AIMS: To study patterns of reserve lipid biosynthesis and turnover (degradation) in two oleaginous Zygomycetes, namely Cunninghamella echinulata and Mortierella isabellina under various growth conditions. Fatty acid composition of the reserve lipid of both strains was also studied in all growth steps. METHODS AND RESULTS: Cunninghamella echinulata and Mortierella isabellina were grown in carbon-excess batch cultures. In the investigated strains, accumulation of reserve lipid occurred only when the activity of both NAD(+)-isocitrate dehydrogenase (ICDH) and NADP(+)-ICDH were not detectable in the cell-free extract. Specifically, in C. echinulata, NAD(+)-ICDH activity was detected even after depletion of ammonium nitrogen in the medium, resulting in a delay of the initiation of lipid accumulation period. On the contrary, in M. isabellina, lipid accumulation occurred simultaneously with ammonium nitrogen exhaustion in the growth medium, as the activity of both NAD(+)- and NADP(+)-ICDH were not detectable after nitrogen depletion. In C. echinulata reserve lipid was not degraded after glucose had been exhausted. Supplementations of the medium with Fe(3+), yeast extract or Mg(2+) induced, however, reserve lipid breakdown and formation of lipid-free material. In M. isabellina after glucose exhaustion, notable lipid degradation occurred, accompanied by a significant lipid-free material biosynthesis. Nevertheless, in multiple-limited media, in which Mg(2+) or yeast extract, besides carbon and nitrogen, were limiting nutrients, reserve lipid breakdown was repressed. In both strains, the quantity of gamma-linolenic acid (GLA) in the reserve lipids [varying between 9 and 16% (w/w) in C. echinulata and 1.5-4.5% (w/w) in M. isabellina] was proportional to lipid-free biomass. CONCLUSIONS: Lipid accumulation period in Zygomycetes is initiated by the attenuation of ICDH activity in the mycelium while the regulation of ICDH from ammonium nitrogen is strain specific. While a single nitrogen limitation was enough to induce lipid accumulation, however, multiple limitations were needed in order to repress lipid turnover in oleaginous Zygomycetes. As for GLA, its biosynthesis in the mycelium seemed proportional to lipid-free biomass synthesis. SIGNIFICANCE AND IMPACT OF THE STUDY: Several nutrients are indispensable for functioning the mechanisms involved in the mobilization of reserve lipid in oleaginous moulds. Therefore, reserve lipid turnover in oleaginous moulds could be repressed in multiple-limited media.

Low-temperature brewing using yeast immobilized on dried figs.

Dried figs, following exhaustive extraction of their residual sugars with water, were used for immobilization of Saccharomyces cerevisiae AXAZ-1. The immobilized biocatalyst was used in repeated batch fermentations of glucose at 30 degrees C, where significant reduction of the fermentation time was observed, falling from 65 h in the first batch to 7 h after the sixth batch. Repeated fermentations of wort at room and low temperatures resulted in fermentation times that fell from 26 to 20 h and from 27 to 24 days at 18 and 3 degrees C, respectively. Ethanol and beer productivities were high, showing suitability of the biocatalyst for low-temperature brewing. Diacetyl concentrations were low (0.3-0.5 mg/L), and polyphenols were lower than in commercial products and decreased as the fermentation temperature was decreased (126-50 mg/L). Ethyl acetate concentrations increased from 53 to 88 mg/L as the temperature was decreased, while the concentration of amyl alcohols at 3 degrees C (58 mg/L) was lower than half of that at 18 degrees C (125 mg/L). The beers produced at the end of the main fermentation had a fine clarity and a special fruity figlike aroma and taste, distinct from commercial products and more intense than beers produced by cells immobilized on other food-grade supports (gluten pellets or delignified cellulosic materials). GC-MS analysis did not show significant differences in the qualitative composition of the aroma compounds of the beers produced by immobilized and free cells.

Production of gamma-linolenic acid by Cunninghamella echinulata cultivated on glucose and orange peel.

A newly isolated strain of Cunninghamella echinulata grown on glucose produced significant quantities of biomass and cellular lipids in media with high C/N ratio. The oil yield from glucose consumed increased after nitrogen exhaustion in the growth medium, but gamma-linolenic acid (GLA) content in cellular oil systematically decreased during the lipid accumulation process. When lipid accumulation was completed, GLA concentration in the cellular lipids progressively increased. The highest GLA production (720 mg/l) was achieved in medium with a C/N ratio equal to 163. C. echinulata was also able to grow on orange peel. The C/N ratio in the orange peel decreased from 50 to 26 during solid-state fermentation. Maximum oxygen uptake was observed during assimilation of reducing sugars, whereas a polygalacturonase activity was detected after reducing sugars had been exhausted. The maximum GLA production was 1.2-1.5 mg/g of fermented peel, calculated on a dry weight basis. After enrichment of the pulp with inorganic nitrogen and glucose, an increase in the production of oil and GLA was observed.

Single cell oil production by Yarrowia lipolytica growing on an industrial derivative of animal fat in batch cultures.

The growth of an oleaginous strain of Yarrowia lipolytica on an industrial fat composed of saturated free fatty acids (stearin) was studied. Lipid accumulation during primary anabolic growth was critically influenced by the medium pH and the incubation temperature. This process was independent of the nitrogen concentration in the culture medium, but was favored at a high carbon substrate level and at a low aeration rate. At pH 6 and a temperature of 28-33 degrees C, 9-12 g/l of dry biomass was produced, whereas significant quantities of lipids were accumulated inside the yeast cells (0.44-0.54 g of lipid per gram of biomass). The strain showed the tendency to degrade its storage lipids, although significant amounts of substrate fat, rich in stearic acid, remained unconsumed in the culture medium. Y. lipolytica presented a strong fatty acid specificity. The fatty acids C12:0, C14:0, and C16:0 were rapidly incorporated and mainly used for growth needs, while C18:0 was incorporated with reduced rates and was mainly accumulated as storage material. Reserve lipids, principally composed of triacylglycerols (55% w/w of total lipids) and free fatty acids (35% w/w), were rich in stearic acid (80% w/w), while negligible amounts of unsaturated fatty acids were detected. When industrial glycerol was used as co-substrate, together with stearin, unsaturated fatty acid concentration in the reserve lipid increased.

Wine production using yeast immobilized on apple pieces at low and room temperatures.

A biocatalyst was prepared by immobilization of Saccharomyces cerevisiae strain AXAZ-1 on apple pieces. It was examined by electron microscope and studied during the fermentation of grape must for batch wine-making. The immobilized yeast showed an important operational stability without any decrease of its activity even at low temperatures (1-12 degrees C). Specifically, at 6 degrees C the biocatalyst favored wine production within 8 days, which is less time than is required for the natural fermentation of grape must. At 1 degrees C wine production was effected in 1 month. This speeding up of the fermentation could be accepted and adopted by the industry for scaling up the wine-making process. Total and volatile acidities were similar to those found in dry wines. The concentrations of higher alcohols (propanol-1 and isobutyl alcohol) were low. The presence of amyl alcohols proved to be temperature dependent and decreased with the temperature decrease. The values of ethyl acetate concentrations were relatively high, up to 154 mg/L. This probably contributes to the fruity aroma and excellent taste of the produced wines. There was no indication of vinegar odor in the product given that the volatile acidity was <0.47 g of acetic acid/L. From the GC-MS analysis it was concluded that cell immobilization did not create serious changes in the product (wine) with regard to the qualitative composition of the aroma compounds.

Effect of aqueous extracts of some plants of Lamiaceae family on the growth of Yarrowia lipolytica.

Aqueous extracts of some plants belonging to the Lamiaceae family (Sideritis montana, Origanum dictamnus, Mentha piperita, Rosmarinus officinallis and Origanum marjorana) caused an important increase of the lag time of Yarrowia lipolytica. Especially, Origanum dictamnus and Rosmarinus officinallis extracts enhanced the lag time considerably and influenced negatively the specific growth rate of this yeast. In culture media having low C/N ratio, all plant extracts caused an increase of the biomass produced in relation to glucose and nitrogen consumed, while, in high C/N ratio media the effect of the extracts on biomass production was negative. In the presence of aqueous plant extracts, in low C/N ratio culture media, the ratio sigma unsaturated/sigma saturated fatty acids in the cellular lipids increased, whereas in high C/N ratio media it decreased.

Kinetic profile of the cellular lipid composition in an oleaginous Yarrowia lipolytica capable of producing a cocoa-butter substitute from industrial fats.

Cell growth, lipid accumulation and cellular lipid composition of Yarrowia lipolytica growing on mixtures of industrial fats containing stearic, oleic, linoleic and palmitic acid have been studied. During growth, the strain incorporated oleic and linoleic acids more rapidly than the saturated fatty acids. Relatively high lipid accumulation (up to 0.44 g of lipids per g of dry matter) was observed when stearic acid was included in the culture medium. In contrast, substrates rich in oleic acid did not favor cellular lipid accumulation. The accumulated lipids, mainly composed of triacylglycerols (45-55% w/w), demonstrated a different total fatty acid composition compared with that of the substrate; in all cases, the microorganism showed the unusual capacity to increase its cellular stearic acid level, even if this fatty acid was not found in high concentrations in the substrate. This permitted the synthesis of interesting lipid profiles with high percentages of stearic acid and non-negligible percentages of palmitic and oleic acid, with a composition resembling that of cocoa-butter.

Continuous wine making by gamma-alumina-supported biocatalyst: quality of the wine and distillates.

The main objective of the present work was the removal of aluminum from wines produced by gamma-alumina-supported yeast cells. Reagents such as Na2CO3, NH4OH, albumin, and Ca(OH)2 were used. Calcium in the presence of albumin was effective, whereas other reagents were not so effective. Because of the improved aroma and taste of distillates produced by gamma-alumina-supported biocatalyst, volatile byproducts of distillates were analyzed. They were also assessed by sensory tests. Methanol, acetaldehyde, ethyl acetate, propanol-1, isobutyl alcohol, and amyl alcohols were determined in distillates. It was noted that the amounts of higher alcohols and amyl alcohols decreased as the temperature of fermentation dropped, leading to a product of improved quality and reduced toxicity.

Effect of a Teucrium polium L. extract on the growth and fatty acid composition of Saccharomyces cerevisiae and Yarrowia lipolytica.

Aqueous Teucrium polium extract slightly inhibits the growth of Saccharomyces cerevisiae (Ki = 0.029 [g/l]-1) and Yarrovia lipolytica (Ki = 0.06l [g/l]-1). However, this extract causes important changes in the unsaturation degree (delta/mol) of the cellular lipids. It moreover favours the increase of the linolenic acid concentration and the decrease of the oleic one in both species.

Microbial fatty acid specificity.

Strains ofRhodotorula sp.,Candida spp. andLangermania sp. cultivated on polyunsaturated oil preferentially incorporated more unsaturated fatty acids. These fatty acids were used mainly for growth needs whereas the saturated ones accumulated in the microbial cell. The cellular oil and the remaining oil in the culture had a lower degree of unsaturation as compared to the initial oil, and a modified fatty acid composition.Candida lipolytica, in a chemostat continuous culture, incorporated C(18) fatty acids in the order of C(18:3)>C(18:2)>C(18:1)>C(18:0), and accumulated mostly the saturated ones. The specific productivity of the cellular oil and of the oil remaining in the culture medium was 0.036 and 0.487 gg(-1) h(-1), respectively, at dilution rateD=0.2/h.