RESUMO
A repetitive batch process was employed followed by membrane ultrafiltration system to produce low-cost cyclodextrins (CDs) using commercial enzymes Toruzyme® cyclomaltodextrin glucanotransferase (CGTase) and its kinetic parameters were determined. The ultrafiltration system enabled the removalof inhibitory products from the reaction medium, allowing the enzyme to be recovered for reuse. A 10 kDa membrane was used to separate the different CDs produced by the CGTase. The substrates evaluated were maltodextrin, corn starch and cassava starch at 5, 10 and 15% (w/V), in the presence and absence of 10% (V/V) ethanol. After reaction for 132 h, 10% (w/V) cassava starch in the presence of ethanol provided the best results with 32.1 mg/mL of ß-CD. Maximum production occurred after 72 h of reaction, with a yield of 87.4% of ß-CD and an α-CD, ß-CD and γ-CD production ratio of 1:1:0.08 g, respectively. When eight repetitive batches of 72 h followed by ultrafiltration and crystallization of ß-CD were performed, 2.1 g of precipitate was obtained with a purity of 67.6% ß-CD. The supernatant from the crystallization process was lyophilized and resulted in 35.3% α-CD. The developed model can be used industrially for the production of low cost CDs from easily obtained raw material
Assuntos
Ultrafiltração/instrumentação , Modelos Econômicos , Tecnologia de Baixo Custo/análise , Ciclodextrinas/farmacologia , Amidos e Féculas , Cristalização/classificaçãoRESUMO
The study comparatively evaluated diverse strategic models of cyclodextrin (CD) production by the CGTase of Bacillus firmus strain 37: continuous production and repetitive batches in ultrafiltration systems; immobilization of CGTase on curdlan and vegetable sponge natural supports; the use of the glycyrrhizin complexing agent to modulate CGTase selectivity in favor of γ-CD production. All strategies had in common the possibility of separation of CGTase from its inhibitory products and its reuse. In the continuous production model, at 48â¯h of assay, the highest productivity and selectivity for ß-CD were obtained, 1.47â¯mmol/L/h and 92.8%, respectively. Glycyrrhizin was able to modulate the production of γ-CD with selectivity of 61.2% for 30-h batches. The comparative evaluation of the different strategic models for obtaining CDs showed particularities that should be considered, and most of the models studied returned satisfactory yields as well as excellent selectivity.
Assuntos
Ciclodextrinas/química , Ciclodextrinas/isolamento & purificação , Enzimas Imobilizadas/química , Enzimas Imobilizadas/metabolismo , Glucosiltransferases/química , Glucosiltransferases/metabolismo , Ultrafiltração/métodos , Bacillus/enzimologia , Compostos Férricos , Fosfatos , Especificidade por SubstratoRESUMO
This study aimed to improve the yield of cyclodextrins (CDs) production in repetitive batches. An innovative ultrafiltration system was used to remove the inhibitory products that accumulated in the medium and to recover the enzyme. The assays were performed with the CGTase from Bacillus firmus strain 37 in purified, semi-purified, and crude extract forms. Maltodextrin (10% w/v) and corn starch (5% w/v) were used as substrates. After eight repetitive 24-h batches, the yield of ß-CD obtained with the purified enzyme and the corn starch substrate was 0.54 mmol/L/h, which was 36% greater than that observed with the 10% maltodextrin substrate. The crude CGTase extract with the corn starch substrate showed a productivity of 0.38 mmol/L/h, which was 29% lower than using the purified enzyme and the corn starch substrate but 7% higher than using the purified enzyme and the maltodextrin substrate. The crude extract, assayed with the corn starch substrate in the presence of 10% ethanol reached 0.43 mmol/L/h productivity, which was 12% higher compared to the assay without ethanol. The semi-purified enzyme was assayed with the corn starch substrate in the presence of 10% ethanol for eight batches lasting 12 h and an excellent selectivity for the ß-CD was obtained, reaching a mean percentage of 96.0%. Therefore, this ultrafiltration system enabled several batches of CD production, with efficient removal of products inhibitory to the CGTase and recovery of the enzyme. The possibility of industrial application of this system is promising.