The use of agriculture substrates in industrial biotechnological processes has been increasing because of its low cost. Cashew apples are considered an agriculture low cost product in the Brazilian Northeast because the cashew cultivation is done mainly to produce cashew nuts. About 90% of the cashew apples production is lost in the field after removing the nut. In this work, the use of clarified cashew apple juice as substrate for microbial cultivation was investigated. The results showed that cashew apple juice is a good source of reducing sugars and can be used to grow Leuconostoc mesenteroides to produce high added value products such as dextran, lactic acid, mannitol and oligosaccharides.

The use of agriculture substrates in industrial biotechnological processes has been increasing because of their low cost. In this work, the use of clarified cashew apple juice was investigated as substrate for enzyme synthesis of prebiotic oligosaccharide. The results showed that cashew apple juice is a good source of reducing sugars and can be used as substrate for the production of dextransucrase by Leuconostoc citreum B-742 for the synthesis of oligosaccharides using the crude enzyme. Optimal oligosaccharide yield (approximately 80%) was obtained for sucrose concentrations lower than 60 g/L and reducing sugar concentrations higher than 100 g/L.

Prebiotic oligosaccharides are nondigestible carbohydrates that can be obtained by enzymatic synthesis. Glucosyltransferases can be used to produce these carbohydrates through an acceptor reaction synthesis. When maltose is the acceptor a trisaccharide composed of one maltose unit and one glucose unit linked by an alpha-1,6-glycosidic bond (panose) is obtained as the primer product of the dextransucrase acceptor reaction. In this work, panose enzymatic synthesis was evaluated by a central composite experimental design in which maltose and sucrose concentration were varied in a wide range of maltose/sucrose ratios in a batch reactor system. A partially purified enzyme was used in order to reduce the process costs, because enzyme purification is one of the most expensive steps in enzymatic synthesis. Even using high maltose/sucrose ratios, dextran and higher-oligosaccharide formation were not avoided. The results showed that intermediate concentrations of sucrose and high maltose concentration resulted in high panose productivity with low dextran and higher-oligosaccharide productivity.

The effects of thermal (pasteurization and sterilization) and non-thermal (ultrasound and plasma) processing on the composition of prebiotic and non-prebiotic acerola juices were evaluated using NMR and GC-MS coupled to chemometrics. The increase in the amount of Vitamin C was the main feature observed after thermal processing, followed by malic acid, choline, trigonelline, and acetaldehyde. On the other hand, thermal processing increased the amount of 2-furoic acid, a degradation product from ascorbic acid, as well as influenced the decrease in the amount of esters and alcohols. In general, the non-thermal processing did not present relevant effect on juices composition. The addition of prebiotics (inulin and gluco-oligosaccharides) decreased the effect of processing on juices composition, which suggested a protective effect by microencapsulation. Therefore, chemometric evaluation of the H qNMR and GC-MS dataset was suitable to follow changes in acerola juice under different processing.

The use of agriculture excess as substrate in industrial fermentations became an interesting alternative to reduce production costs and to reduce negative environmental impact caused by the disposal of these products. In this work, a kinetic study of mannitol production using cashew apple juice as substrate was studied. The carbohydrates of cashew apple juice are glucose and fructose. Sucrose addition favored the yield of mannitol (85%) at the expense of lower productivity. The best results were obtained applying only cashew apple juice as substrate, containing 50 g L(-1) of total reducing sugar (28 g L(-1) of fructose), yielding 18 g L(-1) of mannitol with 67% of fructose conversion into mannitol and productivity of 1.8 g L(-1) h(-1).

The enzyme dextransucrase in a medium containing sucrose and an acceptor as substrate synthesizes prebiotics oligosaccharides. The cashew apple juice works as a source of acceptors because it is rich in glucose and fructose (enzyme acceptors). The use of cashew apple juice becomes interesting because it aims at harnessing the peduncle of the cashew that is wasted during the nut processing, which is the product of greater economic expression. The production of dextransucrase enzyme was done by fermentative process by inoculating the bacterium Leuconostoc mesenteroides NRRL B512F into a culture medium containing sucrose as the only carbon source. Thus, the aim of this work was the production of prebiotic oligosaccharides by enzymatic process with addition of the dextransucrase enzyme to the clarified cashew apple juice. Dextran yield was favored by the combination of low concentrations of sucrose and reducing sugars. The formation of oligosaccharides was favored by increasing the concentration of reducing sugars and by the combination of high concentrations of sucrose and reducing sugars, the highest concentration of oligosaccharides obtained was 104.73 g/L and the qualitative analysis showed that at concentrations of 25 g/L and 75 g/L of sucrose and reducing sugar, respectively, it is possible to obtain oligosaccharides of degree of polymerization up to 12. The juice containing prebiotic oligosaccharide is a potential new functional beverage.

ResumoO presente trabalho teve como objetivo estudar a utilização de excedentes agrícolas como substrato para produção de dextrana e ácido lático. As fermentações foram conduzidas com o microorganismo Leuconostoc mesenteroides B512F, em meio contendo suco de caju e sacarose. As concentrações de açúcar redutor e sacarose foram variadas de acordo com um planejamento experimental. No final da fermentação foram quantificados a dextrana, o ácido lático e a biomassa produzida. Os resultados foram avaliados através da metodologia de análise de superfície de resposta. De acordo com os resultados obtidos, a elevação da concentração de açúcares favorece a produção de dextrana e de ácido lático. A utilização do suco de caju como substrato alternativo para produção de dextrana e ácido lático apresentou viabilidade técnica. Palavras-chave: processos fermentativos; substratos alternativos; ácido lático; suco de caju. AbstractThe main aim of the present work was to study the use of agriculture excess as substrates for dextran and lactic acid production. The fermentations were carried out with the microorganism Leuconostoc mesenteroides B512F in a medium containing cashew apple juice and sucrose. The concentrations of reducing sugar and sucrose were varied according to factorial planning. At the end of the fermentation the dextran, the lactic acid and biomass produced were quantified.The results were analyzed by the surface response analysis methodology. According to the results increasing the sugar level favors dextran and lactic acid production. The use of cashew apple juice as an alternative substrate for dextran and lactic acid production presented technical viability.

Glucosyltransferases can be applied in the synthesis of prebiotic oligosaccharides. Enzymatic synthesis using acceptors can be used to obtain these carbohydrates. When maltose is the acceptor, oligosaccharides containing one maltose moiety and up to eight glucose units linked by a-1,6glycosidic bonds are obtained as the product of dextransucrase acceptor reaction. In this work, the enzymatic synthesis of isomalto-oligosaccharides using dextransucrase from Leuconostoc mesenteroides NRRL B-512F was optimized by response surface methodology. The effect of maltose and sucrose concentrations on the acceptor reaction was evaluated in a batch reactor system. Partially purified enzyme was used to reduce the enzyme purification cost. The results showed that high sucrose concentrations in conjunction with high maltose levels enhanced the isomalto-oligosaccharide synthesis. A productivity of 42.95 mmol/L.h of isomalto-oligosaccharides was obtained at the optimal operating condition (100 mmol/L of sucrose and 200 mmol/L of maltose).j fbc_222 342..354 CorrespondingOligosaccharides as prebiotic have a large application in food formulations, and their beneficial role in human health have been extensively studied. Although the acceptor mechanism of dextransucrase has already been extensively studied, an industrial process has not been developed yet for enzyme synthesis of isomalto-oligosaccharide. The process studied in this work allows the large-scale preparation of isomalto-oligosaccharide using partially purified enzyme. ENZYMATIC SYNTHESIS OF ISOMALTO-OLIGOSACCHARIDES 344M.C. RABELO ET AL.