Lipase-catalysed esters synthesis of cafestol and kahweol.

Cafestol and kahweol (C&K), two coffee diterpene alcohols with structural similarity which exhibit anticarcinogenic effects, were isolated from green coffee Arabica beans, followed by their lipase-catalysed esterification and purification by preparative high-performance liquid chromatography (HPLC). The isolation and enzymatic synthesis parameters of C&K esters were studied, with the latter optimised by a Central Composite Design; both procedures were monitored by gas chromatography. Scale up and improved isolation conditions resulted in 1.29 g of C&K, with 98% purity from 300 g of green Arabica beans. The highest C&K ester yields were observed using an alcohol:fatty acid molar ratio of 1:5, 73.3 mg mL-1 of CAL-B enzyme, 70 °C and 240 rpm for 3 days in toluene, leading to 85-88% conversion among a variety of tested C&K esters, including n-C14:0-C20:0, C18:1, C18:2 and C18:3.

Hydroxylation of 1,8-cineole by Mucor ramannianus and Aspergillus niger

The monoterpenoid 1,8-cineole is obtained from the leaves of Eucalyptus globulus and it has important biological activities. It is a cheap natural substrate because it is a by-product of the Eucalyptus cultivation for wood and pulp production. In this study, it was evaluated the potential of three filamentous fungi in the biotransformation of 1,8-cineole. The study was divided in two steps: first, reactions were carried out with 1,8-cineole at 1 g/L for 24 h; afterwards, reactions were carried out with substrate at 5 g/L for 5 days. The substrate was hydroxylated into 2-exo-hydroxy-1,8-cineole and 3-exo-hydroxy-1,8-cineole by fungi Mucor ramannianus and Aspergillus niger with high stereoselectivity. Trichoderma harzianum was also tested but no transformation was detected. M. ramannianus led to higher than 99% of conversion within 24 h with a starting high substrate concentration (1 g/L). When substrate was added at 5 g/L, only M. ramannianus was able to catalyze the reaction, but the conversion level was 21.7% after 5 days. Both products have defined stereochemistry and could be used as chiral synthons. Furthermore, biological activity has been described for 3-exo-hydroxy-1,8-cineol. To the best of our knowledge, this is the first report on the use of M. ramannianus in this reaction.

Whole cells in enantioselective reduction of benzyl acetoacetate

The β-ketoester benzyl acetoacetate was enantioselectively reduced to benzyl (S)-3-hydroxybutanoate by seven microorganism species. The best result using free cells was obtained with the yeast Hansenula sp., which furnished 97% ee and 85% of conversion within 24 h. After immobilization in calcium alginate spheres, K.marxianus showed to be more stable after 2 cycles of reaction.

Kinetic modeling of solvent-free lipase-catalyzed partial hydrolysis of palm oil.

This work reports the experimental data and kinetic modeling of diacylglycerol (DAG) production from palm oil using a commercial immobilized lipase (Lipozyme RM IM) in a solvent-free medium. The experiments were performed in batch mode, at 55 °C and 400 rpm, and the effects of enzyme concentration (0.68-2.04 wt% related to the mass of substrates), initial water concentration (5-15 wt% related to the mass of oil), and reaction time were evaluated. A novel kinetic model is presented based on the ordered-sequential bi-bi mechanism considering hydrolysis and esterification steps, in which a correlation between water-in-oil solubility and surfactant molecules concentration in the oil allowed the model to describe the induction period in the beginning of the hydrolysis reaction. Moreover, mass transfer limitations related to the enzyme concentration in the system were also taken into account. The proposed model presented a very satisfactory agreement with the experimental data, thus allowing a better understanding of the reaction kinetics. The best conditions obtained for the product (partially hydrolyzed palm oil) in terms of DAG yield (35.91 wt%) were 2.87 wt% enzyme/substrate, 2.10 wt% water/oil, and 72 h of reaction.