Mario Aguedo

Role of β-Oxidation Enzymes in γ-Decalactone Production by the Yeast Yarrowia lipolytica

ABSTRACT Some microorganisms can transform methyl ricinoleate into γ-decalactone, a valuable aroma compound, but yields of the bioconversion are low due to (i) incomplete conversion of ricinoleate (C18) to the C10 precursor of γ-decalactone, (ii) accumulation of other lactones (3-hydroxy-γ-decalactone and 2- and 3-decen-4-olide), and (iii) γ-decalactone reconsumption. We evaluated acyl coenzyme A (acyl-CoA) oxidase activity (encoded by the POX1 throughPOX5 genes) in Yarrowia lipolytica in lactone accumulation and γ-decalactone reconsumption inPOX mutants. Mutants with no acyl-CoA oxidase activity could not reconsume γ-decalactone, and mutants with a disruption ofpox3, which encodes the short-chain acyl-CoA oxidase, reconsumed it more slowly. 3-Hydroxy-γ-decalactone accumulation during transformation of methyl ricinoleate suggests that, in wild-type strains, β-oxidation is controlled by 3-hydroxyacyl-CoA dehydrogenase. In mutants with low acyl-CoA oxidase activity, however, the acyl-CoA oxidase controls the β-oxidation flux. We also identified mutant strains that produced 26 times more γ-decalactone than the wild-type parents.

Involvement of Acyl Coenzyme A Oxidase Isozymes in Biotransformation of Methyl Ricinoleate into γ-Decalactone by Yarrowia lipolytica

ABSTRACT We reported previously on the function of acyl coenzyme A (acyl-CoA) oxidase isozymes in the yeast Yarrowia lipolytica by investigating strains disrupted in one or several acyl-CoA oxidase-encoding genes (POX1 throughPOX5) (H. Wang et al., J. Bacteriol. 181:5140–5148, 1999). Here, these mutants were studied for lactone production. Monodisrupted strains produced similar levels of lactone as the wild-type strain (50 mg/liter) except for Δpox3, which produced 220 mg of γ-decalactone per liter after 24 h. The Δpox2 Δpox3 double-disrupted strain, although slightly affected in growth, produced about 150 mg of lactone per liter, indicating that Aox2p was not essential for the biotransformation. The Δpox2 Δpox3 Δpox5 triple-disrupted strain produced and consumed lactone very slowly. On the contrary, the Δpox2 Δpox3 Δpox4 Δpox5 multidisrupted strain did not grow or biotransform methyl ricinoleate into γ-decalactone, demonstrating that Aox4p is essential for the biotransformation.

Application of a novel oscillatory flow micro-bioreactor to the production of γ-decalactone in a two immiscible liquid phase medium

A novel micro-bioreactor based on the oscillatory flow technology was applied to the scale-down of the biotechnological production of γ-decalactone. A decrease up to 50% of the time required to obtain the maximum concentration of the compound was observed, when compared with other scaled-down platforms (stirred tank bioreactor or shake flask). A three-fold increase in γ-decalactone productivity was obtained by increasing oscillatory mixing intensity from Reo ~482 to Reo ~1447. This was presumably related to the effective contribution of the reactor geometry to enhanced mass transfer rates between the two immiscible liquid phases involved in the process by increasing the interfacial area.

Impact of lignin structure on oil production via hydroprocessing with a copper-doped porous metal oxide catalyst

A copper-catalyzed depolymerization strategy was employed to investigate the impact of lignin structure on the distribution of hydroprocessing products. Specifically, lignin was extracted from beech wood and miscanthus grass. The extracted lignins, as well as a commercial lignin (P1000), were then fractionated using ethyl acetate to provide three different portions for each source of lignin [total of 9 fractions]. Each fraction was structurally characterized and treated with a copper-doped porous metal oxide (Cu-PMO) catalyst under 4MPa H2 and at 180°C for 12h. The reaction conditions provided notable yields of oil for each fraction of lignin. Analysis of the oils indicated phenolic monomers of commercial interest. The structure of these monomers and the yield of monomer-containing oil was dependent on the origin of the lignin. Our results indicate that hydroprocessing with a Cu-PMO catalyst can selectively provide monomers of commercial interest by careful choice of lignin starting material.

Effect of Organic Solvents on Microalgae Growth, Metabolism and Industrial Bioproduct Extraction: A Review

In this review, the effect of organic solvents on microalgae cultures from molecular to industrial scale is presented. Traditional organic solvents and solvents of new generation-ionic liquids (ILs), are considered. Alterations in microalgal cell metabolism and synthesis of target products (pigments, proteins, lipids), as a result of exposure to organic solvents, are summarized. Applications of organic solvents as a carbon source for microalgal growth and production of target molecules are discussed. Possible implementation of various industrial effluents containing organic solvents into microalgal cultivation media, is evaluated. The effect of organic solvents on extraction of target compounds from microalgae is also considered. Techniques for lipid and carotenoid extraction from viable microalgal biomass (milking methods) and dead microalgal biomass (classical methods) are depicted. Moreover, the economic survey of lipid and carotenoid extraction from microalgae biomass, by means of different techniques and solvents, is conducted.

Hydrothermal Pretreatments of Macroalgal Biomass for Biorefineries

Recently, macroalgal biomass is gaining wide attention as an alternative in the production of biofuels (as bioetanol and biogas) and compounds with high added value with specific properties (antioxidants, anticoagulants, anti-inflammatories) for applications in food, medical and energy industries in accordance with the integrated biorefineries. Furthermore, biorefinery concept requires processes that allow efficient utilization of all components of the biomass. The pretreatment step in a biorefinery is often based on hydrothermal principles of high temperatures in aqueous solution. Therefore, in this chapter, a review on the application of hydrothermal pretreatment on macroalgal biomass is presented.

Structure impact of two galactomannan fractions on their viscosity properties in dilute solution, unperturbed state and gel state

Two fractions of carob galactomannans (GM25 and GM80) were extracted at respectively 25°C and 80°C from crude locust bean gum. Those fractions having slightly different chemical structures, previously characterized, were studied for their viscosity properties over a wide range of concentrations: diluted solution, unperturbed state and gel state. For each of the physical properties, links to the chemical fine structure could be established, expanding knowledge on the topic: in dilute solution, GM25 is more soluble in water while GM80 seems to tend to self-association due to its structure as highlighted by intrinsic viscosity measurements ([η]GM25=9.96dLg-1 and [η]GM80=4.04dLg-1). In unperturbed state, initial viscosities η0 were more important for GM80 fractions at 1% and 2% due to greater hyperentanglements (η0(GM80,1%)=9.9Pas; η0(GM80,2%)=832.0; Pa.s η0(GM25,1%)=3.1Pas; η0(GM25,2%)=45.1Pas). In gel state, hydrogels obtained from GM80 were also stronger (hardness GM80 (2%)=0.51N and hardness GM25 (2%)=0.11N), suggesting a much more important number of junction areas within the gel network. The findings discussed herein demonstrate the potential for new applications.

Lignin fractionation as an efficient route for enhancing Polylactide thermal stability and flame retardancy

Abstract Due to its variable botanical origin, functionality, reactivity, and heterogeneity, using lignin in industrial application is not an easy task. In the present study, we investigate the effect of lignin fractionation as a simple way for reducing the variability in its properties. Kraft lignin was separated by ultrafiltration membranes in two fractions with a specific molecular weight and the properties of each fraction were characterized through FTIR, XPS, TGA and cone calorimeter test. Lignin fractions display different thermal and combustion behaviors. Thus, the two fractions have been evaluated as flame retardant additives for polylactide (PLA). PLA composites, containing well dispersed lignin (20 wt%), were produced by melt blending in an internal mixer. The thermo-degradant effect of each fraction on PLA during melt processing was investigated by rheological analysis and size exclusion chromatography while the composites thermal stability and fire properties were evaluated using TGA and cone calorimeter test. Results showed that using appropriate lignin fraction enables for obtaining PLA composites presenting enhanced properties

Preliminary characterisation of residual biomass from Hibiscus sabdariffa calyces

Hibiscus sabdariffa calyces are mainly used for different agro-food and beverages applications. The residual biomass generated contains various useful substances that were extracted and characterized. It contained 23% (w/w) soluble pectic material, a food additive, extracted with hot acidified water (80°C, pH = 1.5) and precipitated with ethanol. The molecular weight (28.5 and 109.7 kDa), the degree of methylation (70.6 and 44.3%) and the degree of acetylation (19.0 and 4.9%) were determined for two Senegalese cultivars (koor and vimto, respectively). The effect of the extraction method on these parameters was highlighted. The residual lignocellulosic material (LCM) was chemically degraded to monosaccharides and the amount of glucose and xylose (39% of dry LCM) determined to estimate its potential as feedstock for biofuels production. However, an enzymatic degradation test revealed a recalcitrant LCM, as only 50 to 55% of its polymeric glucose content was degraded to monosaccharides without pretreatment. Xylo-oligosaccharides (XOS) are functional foods with a real market potential as prebiotics, characterized by their degree of polymerization (DP). The production of XOS synthetized by the enzymatic degradation of LCM was monitored. The results of analyses performed showed that XOS produced had mainly DP3 and DP4 values. Key words : Pectin, lignocellulosic material, enzymatic degradation, xylo-oligosaccharides.