β-Fructofuranosidase production by repeated batch fermentation with immobilized Aspergillus japonicus

Abstract

The fungus Aspergillus japonicus ATCC 20236 was immobilized in vegetal fiber and used in repeated batch fermentations of sucrose (200 g/l) for the production of beta-fructofuranosidases (FFase). The assays were performed during eight consecutive cycles that were completed in a total period of 216 h. After each 24-h cycle of fermentation (except for the first cycle, which lasted 48 h), the fermented broth was replaced by fresh medium, and the FFase activity was determined in the replaced medium. The average value of FFase activity was a constant 40.6 U/ml at the end of the initial seven cycles, but had decreased by 22% at the end of the eighth cycle. Concurrent with these high and constant FFase values, the hydrolyzing activity of this enzyme increased during the cycles, while the transfructosylating activity decreased. As a consequence, the maximum production of fructooligosaccharides of 134.60 g/l observed in the initial 30 h of fermentation (first cycle) had gradually decreased by the end of the subsequent cycles, reaching approximately 23% of this value during cycles 4-8. Based on these results, we conclude that the present immobilization system has a great potential for application in a semi-continuous process for the production of FFase, but further studies are necessary to maintain the FFase transfructosylation activity at high levels during the overall process.

Production of fructooligosaccharides and β-fructofuranosidase by batch and repeated batch fermentation with immobilized cells of Penicillium expansum

“…3) that is a very interesting result due to the industrial importance of this enzyme. Similar results were obtained by Mussatto et al [9] during the FOS production by A. japonicus in repeated batch fermentations, where the synthesis of this oligosaccharide decreased along the cycles, while the FFase activity remained constant. Nevertheless, the FOS production by repeated batch fermentation with P. expansum (present study) gave better results than our previous study of FOS production by repeated batch fermentation with A. japonicus immobilized on SF [9], revealing the greater ability of P. expansum for use on the production of FOS.…”

“…FOS (1-kestose, 1-nystose, and 1-b-fructofuranosyl nystose) and residual sugars (sucrose, fructose, and glucose) were analyzed using high-performance liquid chromatography (HPLC) [9] on an equipment LC-10 A (Jasco, Japan) with a Prevail Carbohydrate ES column (5 lm, 250 9 4.6 mm, Alltech) at room temperature and a refractive index detector. The response of the refractive index detector was recorded and integrated using the Star Chromatography Workstation software (Varian).…”

“…The b-fructofuranosidase (FFase) activity was determined by quantification of the glucose released from sucrose, as previously described [9]. The reaction mixture contained 100 ml of the crude FFase extract, 300 mmol of sucrose and 50 mmol of sodium acetate buffer (pH 5.0) in a total volume of 1 ml.…”

“…Our research group has been focused in finding an efficient process for FOS and b-fructofuranosidase (FFase, enzyme with hydrolysis and transfructosylation activity) production able to be used in industrial scale [8][9][10]. Although several Aspergillus japonicus strains have been reported as potentially adequate for industrial production of FOS [11], Penicillium expansum was recently found as being a fungal strain of great potential for FOS production from sucrose [12].…”

Morphology engineering of Aspergillus niger for improved enzyme production

“…As shown, the strategy could be successfully transferred to a fructofuranosidase production process, where a final titer of 160 U/mL could be obtained. This value is about fourfold higher as compared to titers obtained until recently (Mussatto et al, 2009), underlining the enormous potential of the novel microparticle-based approach. Taking also the first successful example of chloroperoxidase production by the filamentous fungus C. fumago (Kaup et al, 2007) into account, the enhancing effects of microparticles seem of general nature and thus of high relevance for biotechnological processes with filamentous microorganisms.…”