RESUMEN
Deep Eutectic Solvents (DES) are used as reaction media for lipase-catalyzed esterifications in continuous devices. In particular, DES may be useful for lipophilization-like reactions involving substrates with unpaired solubilities. Aspects to be considered are the viscosity of the solvent, as well as the stability of the enzyme in the non-conventional media. The viscosity can be decreased by adding buffer as cosolvent (up to 20% v/v) and keeping the non-conventional nature. Lipases can be stabilized by following a double immobilization pattern, comprising CLEA formation and entrapment in LentiKats®. The low viscosity and high stability of the CLEA-LK-lipase enable the use of DES under flow conditions.
Asunto(s)
Disolventes Eutécticos Profundos , Lipasa , Catálisis , Esterificación , Lipasa/metabolismo , Solventes , ViscosidadRESUMEN
In recent years, residual glycerol from biodiesel synthesis made this chemical a cheap, readily available carbon source to bioprocess, which is also a form to reduce costs in the fuel industry. We propose and describe a bioprocess using fluidized and packed-bed continuous bioreactors to convert this residual glycerol into value-added products such as 1,3-propanediol (1,3-PD) and 2,3-butanediol (2,3-BD), largely used in the chemical industry. The bacterium Klebsiella pneumoniae BLh-1, strain isolated by us, was immobilized in the permeable support of polyvinyl alcohol (LentiKats®). After testing different dilution rates (D) for all bioreactor configurations, the best obtained productivities of 1,3-PD was 8.69 g L-1 h-1 at a D = 0.45 h-1 , and 2.99 g L-1 h-1 at a D = 0.30 h-1 for 2,3-BD, both in the packed-bed configuration. In the fluidized-bed reactor, the highest productivity values achieved were 4.48 and 1.16 g L-1 h-1 for 1,3-PD and 2,3-BD, respectively, both at D = 0.33 h-1 . These results show the potential of setting up a bioprocess based on continuous cultures using immobilized K. pneumoniae BLh-1 in PVA matrices in order to efficiently convert the abundant surplus of glycerol into commercially important chemicals such as 1,3-PD and 2,3-BD.
Asunto(s)
Glicerol , Klebsiella pneumoniae , Biocombustibles , Reactores Biológicos , Butileno Glicoles , Glicoles de PropilenoRESUMEN
We investigated the fermentation of a mixture of oat and soybean hulls (1:1) subjected to acid (AH) or enzymatic (EH) hydrolyses, with both showing high osmotic pressures (> 1200 Osm kg-1) for the production of ethanol. Yeasts of genera Spathaspora, Scheffersomyces, Sugiymaella, and Candida, most of them biodiverse Brazilian isolates and previously untested in bioprocesses, were cultivated in these hydrolysates. Spathaspora passalidarum UFMG-CM-469 showed the best ethanol production kinetics in suspended cells cultures in acid hydrolysate, under microaerobic and anaerobic conditions. This strain was immobilized in LentiKats® (polyvinyl alcohol) and cultured in AH and EH. Supplementation of hydrolysates with crude yeast extract and peptone was also performed. The highest ethanol production was obtained using hydrolysates supplemented with crude yeast extract (AH-CYE and EH-CYE) showing yields of 0.40 and 0.44 g g-1, and productivities of 0.39 and 0.29 g (L h)-1, respectively. The reuse of the immobilized cells was tested in sequential fermentations of AH-CYE, EH-CYE, and a mixture of acid and enzymatic hydrolysates (AEH-CYE) operated under batch fluidized bed, with ethanol yields ranging from 0.31 to 0.40 g g-1 and productivities from 0.14 to 0.23 g (L h)-1. These results warrant further research using Spathaspora yeasts for second-generation ethanol production.
Asunto(s)
Células Inmovilizadas , Etanol , Glycine max/metabolismo , Saccharomycetales , Xilosa/metabolismo , Avena/metabolismo , Biocombustibles/microbiología , Reactores Biológicos/microbiología , Células Inmovilizadas/citología , Células Inmovilizadas/metabolismo , Etanol/análisis , Etanol/metabolismo , Fermentación , Lignina/metabolismo , Saccharomycetales/citología , Saccharomycetales/metabolismoRESUMEN
Laccase from Pycnoporus sanguineus CS43 was successfully immobilized onto Immobead-150 and Eupergit-C by covalent binding and by entrapment in LentiKats. The highest immobilization was onto Immobead-150 (97.1±1.2%) compared to the other supports, LentiKats (89±1.1%) and Eupergit-C (83.2±1.4%). All three immobilized enzyme systems showed increased thermostability and better mechanical properties than free laccase. Moreover, after 5 cycles of reuse of these systems, 90% of initial laccase activity was retained. Immobead-150 and LentiKats systems exhibited the highest efficiencies in removal of m-cresol under the combined actions of biodegradation and adsorption, while laccase entrapped in LentiKats showed a high ability for degradation of m-cresol within 24h. In addition, the typical Michaelis-Menten enzymatic model effectively described the kinetic profile of m-cresol degradation by the enzyme entrapped in LentiKats. Based on the results obtained in the present study, it can be established that the immobilized biocatalysts developed here possess significant potential for wastewater treatment.