In the present report, crude glycerol, waste discharged from bio‐diesel production, was used as carbon substrate for three natural Yarrowia lipolytica strains (LFMB 19, LFMB 20 and ACA‐YC 5033) during growth in nitrogen‐limited submerged shake‐flask experiments. In media with initial glycerol concentration of 30 g/L, all strains presented satisfactory microbial growth and complete glycerol uptake. Although culture conditions favored the secretion of citric acid (and potentially the accumulation of storage lipid), for the strains LFMB 19 and LFMB 20, polyol mannitol was the principal metabolic product synthesized (maximum quantity 6.0 g/L, yield 0.20–0.26 g per g of glycerol consumed). The above strains produced small quantities of lipids and citric acid. In contrast, Y. lipolytica ACA‐YC 5033 produced simultaneously higher quantities of lipid and citric acid and was further grown on crude glycerol in nitrogen‐limited experiments, with constant nitrogen and increasing glycerol concentrations (70–120 g/L). Citric acid and lipid concentrations increased with increment of glycerol; maximum total citric acid 50.1 g/L was produced (yield 0.44 g per g of glycerol) while simultaneously 2.0 g/L of fat were accumulated inside the cells (0.31 g of lipid per g of dry weight). Cellular lipids were mainly composed of neutral fraction, the concentration of which substantially increased with time. Moreover, in any case, the phospholipid fraction was more unsaturated compared with total and neutral lipids, while at the early growth step, microbial lipid was more rich in saturated fatty acids (e.g. C16:0 and C18:0) compared with the stationary phase.

The last years there has been a significant rise in the number of publications in the international literature that deal with the production of lipids by microbial sources (the 'single cell oils; SCOs' that are produced by the so-called 'oleaginous' micro-organisms). In the first part of the present review article, a general overview of the oleaginous micro-organisms (mostly yeasts, algae and fungi) and their potential upon the production of SCOs is presented. Thereafter, physiological and kinetic events related with the production of, mostly, yeast and fungal lipids when sugars and related substrates like polysaccharides, glycerol, etc. (the de novo lipid accumulation process) or hydrophobic substrates like oils and fats (the ex novo lipid accumulation process) were employed as microbial carbon sources, are presented and critically discussed. Considerations related with the degradation of storage lipid that had been previously accumulated inside the cells, are also presented. The interplay of the synthesis of yeast and fungal lipids with other intracellular (i.e. endopolysaccharides) or extracellular (i.e. citric acid) secondary metabolites synthesized is also presented. Finally, aspects related with the lipid extraction and lipidome analysis of the oleaginous micro-organisms are presented and critically discussed.

Aims:  In this study, we have investigated the biochemical behaviour of Aspergillus sp. (five strains) and Penicillium expansum (one strain) fungi cultivated on waste cooking olive oil. The production of lipid‐rich biomass was the main target of the work. In parallel, the biosynthesis of other extracellular metabolites (organic acids) and enzyme (lipase) and the substrate fatty acid specificity of the strains were studied. Methods and Results:  Carbon‐limited cultures were performed on waste oil, added in the growth medium at 15 g l−1, and high biomass quantities were produced (up to c. 18 g l−1, conversion yield of c. 1·0 g of dry biomass formed per g of fat consumed or higher). Cellular lipids were accumulated in notable quantities in almost all cultures. Aspergillus sp. ATHUM 3482 accumulated lipid up to 64·0% (w/w) in dry fungal mass. In parallel, extracellular lipase activity was quantified, and it was revealed to be strain and fermentation time dependent, with a maximum quantity of 645 U ml−1 being obtained by Aspergillus niger NRRL 363. Storage lipid content significantly decreased at the stationary growth phase. Some differences in the fatty acid composition of both cellular and residual lipids when compared with the initial substrate fat used were observed; in various cases, cellular lipids more saturated and enriched with arachidic acid were produced. Aspergillus strains produced oxalic acid up to 5·0 g l−1. Conclusions:  Aspergillus and Penicillium strains are able to convert waste cooking olive oil into high‐added‐value products. Significance and Impact of the Study:  Increasing fatty wastes amounts are annually produced. The current study provided an alternative way of biovalourization of these materials, by using them as substrates, to produce added‐value compounds.

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The influence of environmental parameters on mycelial linear growth of Pleurotus ostreatus, P. eryngii, P. pulmonarius, Agrocybe aegerita, Lentinula edodes, Volvariella volvacea and Auricularia auricula-judae was determined in two different nutrient media in a wide range of temperature, forming the basis for the assessment of their temperature optima. V. volvacea grew faster at 35 degrees C, P. eryngii at 25 degrees C, P. ostreatus and P. pulmonarius at 30 degrees C, A. aegerita at 25 or 30 degrees C and A. auricula-judae at 20 or 25 degrees C depending on the nutrient medium used and L. edodes at 20 or 30 degrees C depending on the strain examined. The mycelium extension rates were evaluated on seven mushroom cultivation substrates: wheat straw, cotton gin-trash, peanut shells, poplar sawdust, oak sawdust, corn cobs and olive press-cake. The mycelium extension rates (linear growth and colonization rates) were determined by the 'race-tube' technique, and were found to be the highest on cotton gin-trash, peanut shells and poplar sawdust for Pleurotus spp. and A. aegerita. Wheat straw, peanut shells and particularly cotton gin-trash supported fast growth of V. volvacea, whereas wheat straw was the most suitable substrate for L. edodes and A. auricula-judae. Supplemented oak sawdust and olive press-cake were poor substrates for most species examined, while almost all strains performed adequately on corn cobs.

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Thamnidium elegans CCF-1465 was grown in a submerged shake-flask culture in media in which inexpensive sugars derived from a sugar refinery plant (glucose, fructose, and sucrose) were utilized as carbon sources. Cultures were performed in media favoring the accumulation of reserve lipid (high initial C/N ratios imposed, media presenting constant initial nitrogen and increasing initial sugar concentrations, namely, at 30, 60, and 80 g/L). Although significant quantities of fat were accumulated in all cultures performed, lipid production in terms of both absolute (g/L) and relative (%, wt/wt, in dry matter) values clearly increased with sugar concentration increments into the medium. For a given initial sugar concentration, regardless of the individual sugar utilized, lipid was accumulated in almost equivalent quantities inside the mycelia. Sugars were assimilated with comparable consumption rates, while consumption rate was divided into two distinct phases, especially in the high initial sugar concentration media; in the first growth phase, an almost linear assimilation rate of all individual sugars occurred with an uptake rate of ∼0.20 g/L 3 h. Thereafter, (in the lipid accumulation period), a decrease of the sugar uptake rate was observed, that remained linear with a noticeably lower value (∼0.09 g/L 3 h) regardless of the initial sugar concentration or the nature of the sugar used. Maximum lipid quantities of more than 9 g/L, with 70% (w/w) fat in dry microbial mass and conversion yield of more than 20% (w/w) lipid per sugar consumed, were obtained. The nature and concentration of sugar as well as the fermentation time seemed to have some effect upon distribution of cellular fatty acids in the various lipids produced. Microbial lipids principally being composed of the fatty acids oleic and palmitic, therefore constituting perfect starting materials for the production of "2nd generation" biodiesel, were produced regardless of the sugar used as substrate. The medically important γ-linolenic acid was produced in quantities around 510 mg/L. Phospholipids were the more unsaturated fraction of cellular lipids.

Mycelium growth rates, biomass concentration (estimated as glucosamine content) and laccase and endoglucanase secretion were monitored during solid state fermentation (SSF) of wheat straw (WS), reed grass (RG) and bean stalk (BS) residues by Lentinula edodes strains 119, 121, and 122. In a first experiment, these strains were subjected to screening regarding their growth rates and biomass yield, where strain 121 proved to be the fastest colonizer. However, the greater biomass yield at the end of colonization was demonstrated by strain 122 on BS (465.93 mg g -1 d.w.). In a second experiment, growth characters, as well as endoglucanase and laccase production patterns of the selected strains 121 and 122 were monitored at three intervals i.e., at 33, 66, and 100% of substrate colonization. BS furnished the highest endoglucanase production for strain 121, while RG for strain 122. A strain and substrate-dependent behaviour of the enzyme secretion was detected, with strain 122 presenting maximal endoglucanase activity in all substrates at the initial (33%) and final (100%) stages of colonization (0.64-0.90 and 0.79-0.97 U g -1, respectively). However, in strain 121 the peak of endoglucanase production was detected in the early stages of colonization (at 33% on WS and at 66% on RG and BS). Laccase activity showed increased values (maxima on WS, 353.68 and 548.67 U g -1 by strains 121 and 122, respectively) at 66% of colonization. Correlation analysis of growth data demonstrated negative relations between growth rate and biomass yield and between laccase and endoglucanase activities on WS and RG substrates fermented by strain 122. Finally, possible relations of growth parameters with nutritional constituents of the substrates were investigated.