Rodrigo O. M. A. de Souza

Lipases: Valuable catalysts for dynamic kinetic resolutions

Dynamic kinetic resolutions have proven to be a useful method for the preparation of enantiopure compounds from racemates, leading to the formation of a single enantiomer in theoretically 100% yield. Because lipases are ubiquitous, versatile, stereoselective and robust biocatalysts, they have been successfully applied as co-catalysts in these reactions, being mostly combined with metals in the chemoenzymatic dynamic kinetic resolutions of alcohols and amines.

The Three‐Component Biginelli Reaction: A Combined Experimental and Theoretical Mechanistic Investigation

Biginelli reactions have been monitored by direct infusion electrospray ionization mass spectrometry (ESI-MS) and key cationic intermediates involved in this three-component reaction have been intercepted and further characterized by tandem MS experiments (ESI-MS/MS). Density functional theory calculations were also used to investigate the feasibility of the major competing mechanisms proposed for the Biginelli reaction. The experimental and theoretical results were found to corroborate the iminium mechanism proposed by Folkers and Johnson, whereas no intermediates directly associated with either the more energy demanding Knoevenagel or enamine mechanisms could be intercepted.

Kinetic Resolution of rac-1-Phenylethanol with Immobilized Lipases: A Critical Comparison of Microwave and Conventional Heating Protocols∥

The lipase-catalyzed kinetic resolution of rac-1-phenylethanol with vinyl acetate as acyl donor and cyclohexane as solvent has been investigated applying both microwave dielectric heating and conventional thermal heating in order to probe the existence of nonthermal microwave effects. All transformations were conducted at 40 degrees C in a dedicated reactor setup that allowed accurate internal reaction temperature measurements with use of fiber-optic probes. Quartz reaction vessels that allow higher levels of microwave power to be administered to the reaction mixture were used for all experiments. For all five studied immobilized lipases, the observed reactivities and enantioselectivities in microwave and oil bath experiments were identical and thus not related to the presence of the microwave field. The effect of magnetic stirring proved critical as too rapid stirring in some instances destroyed the enzyme support structure and led to altered reactivities and selectivities.

Lipase-catalyzed diacylglycerol production under sonochemical irradiation☆

The present paper describes a protocol for production of diacylglycerol by the partial hydrolysis of soybean oil catalyzed by lipase under ultrasound irradiation. Better yields and shorter reaction times were obtained under sonication as compared to the thermal process.

Bio(chemo)technological strategies for biomass conversion into bioethanol and key carboxylic acids.

Biomass conversion into valuable compounds including fuels, chemicals and materials has become the subject of a wide range of studies in recent years. In spite of the several reports available on transformations of biomass-derived platform molecules into a range of products, chemo-catalytic and biotechnological strategies for the conversion of various feedstocks to key platform chemicals constitute an important area for biomass deconstruction which will allow subsequent processes for high added value products. This contribution has the objective of disclosing a series of strategies for the processing and pretreatment of key biomass feedstocks into platform chemicals, namely bioethanol and key carboxylic acids (e.g. levulinic and succinic acids).

A Retrosynthesis Approach for Biocatalysis in Organic Synthesis

For the planning of an organic synthesis route, the disconnection approach guided by retrosynthetic analysis of possible intermediates and the chemical reactions involved, back to ready available starting materials, is well established. In contrast, such concepts just get developed for biocatalytic routes. In this Review we highlight functional group interconversions catalyzed by enzymes. The article is organized rather by chemical bonds formed-exemplified for C-N, C-O- and C-C-bonds-and not by enzyme classes, covering a broad range of reactions to incorporate the desired functionality in the target molecule. Furthermore, the successful use of biocatalysts, also in combination with chemical steps, is exemplified for the synthesis of various drugs and advanced pharmaceutical intermediates such as Crispine A, Sitagliptin and Atorvastatin. This Review also provides some basic guidelines to choose the most appropriate enzyme for a targeted reaction keeping in mind aspects like commercial availability, cofactor-requirement, solvent tolerance, use of isolated enzymes or whole cell recombinant microorganisms aiming to assist organic chemists in the use of enzymes for synthetic applications.

Palm oil hydrolysis catalyzed by lipases under ultrasound irradiation – The use of experimental design as a tool for variables evaluation

Diacylglycerol oil has been increasingly recognized by its good nutritional properties and therefore, different technologies have been developed for obtaining it. The present work focuses on the diacylglycerol production by hydrolysis reaction of the palm oil using the PS IM and TL IM commercial lipases as biocatalysts under ultrasound irradiation. An experimental design (central composite rotatable design--CCRD) adopting surface response was applied as a tool to evaluate the optimal reaction conditions beyond a restrict number of experiments. Reactions were performed in an ultrasound equipment and different variables were investigated, such as temperature (30-55 °C), enzyme content (1-2 wt.% of oil mass), mechanical stirring (300-700 rpm) and reaction time. Both, PS IM and TL IM enzymes showed the best results after 1h and 30 min of reaction under 30 °C and, applying 300 rpm as stirring. On these conditions, the diacylglycerol yield was around 34% and 39%, respectively; considering that 1% PS IM was applied for the first one and, 2% TL IM for the second one. Therefore, it was obtained good yield of a diacylglycerol-rich oil in shorter reaction times under sonication and soft conditions. The mathematic model proposed suggested a satisfactorily representation of the process and good correlation among the experimental results and the theoretical values predicted by the model equation were achieved.

Continuous flow valorization of fatty acid waste using silica-immobilized lipases

Silica immobilized lipases have been prepared and utilized in the valorization of fatty acid-derived food waste streams under continuous flow conditions. Findings demonstrate that better conversions could be obtained when compared with commercially available immobilized enzymes.

The Multicomponent Hantzsch Reaction: Comprehensive Mass Spectrometry Monitoring Using Charge‐Tagged Reagents

A novel strategy for the ESI-MS monitoring of reaction solutions involving the alternate use of permanently charge-tagged reagents has been used for comprehensive mass spectrometry monitoring of the multicomponent Hantzsch 1,4-dihydropyridine reaction. By placing a charge tag on either, or both, of the two key reactants, ion suppression effects for ESI were eliminated or minimized, and comprehensive detection of charge-tagged intermediates was achieved. The strategy allowed the trapping and characterization of the important intermediates in the mechanism for 1,4-dihydropyridine formation.

Stability and structural changes of horseradish peroxidase: microwave versus conventional heating treatment.

Effects of conventional heating (CH) and microwave (MW) on the structure and activity of horseradish peroxidase (HRP) in buffer solution were studied. CH incubation between 30 and 45 °C increased activity of HRP, reaching 170% of residual activity (RA) after 4-6 h at 45 °C. CH treatment at 50 and 60 °C caused HRP inactivation: RA was 5.7 and 16.7% after 12 h, respectively. Secondary and tertiary HRP structural changes were analyzed by circular dichroism (CD) and intrinsic fluorescence emission, respectively. Under CH, activation of the enzyme was attributed to conformational changes in secondary and tertiary structures. MW treatment had significant effects on the residual activity of HRP. MW treatment at 45 °C/30W followed by CH treatment 45 °C regenerated the enzyme activity. The greatest loss in activity occurred at 60 °C/60 W/30 min (RA 16.9%); without recovery of the original activity. The inactivation of MW-treated HRP was related to the loss of tertiary structure, indicating changes around the tryptophan environment.