Amberlite IRA 900 versus calcium alginate in immobilization of a novel, engineered β‐fructofuranosidase for short‐chain fructooligosaccharide synthesis from sucrose

Abstract

The immobilization of β-fructofuranosidase for short-chain fructooligosaccharide (scFOS) synthesis holds the potential for a more efficient use of the biocatalyst. However, the choice of carrier and immobilization technique is a key to achieving that efficiency. In this study, calcium alginate (CA), Amberlite IRA 900 (AI900) and Dowex Marathon MSA (DMM) were tested as supports for immobilizing a novel engineered β-fructofuranosidase from Aspergillus japonicus for scFOS synthesis. Several immobilization parameters were estimated to ascertain the effectiveness of the carriers in immobilizing the enzyme. The performance of the immobilized biocatalysts are compared in terms of the yield of scFOS produced and reusability. The selection of carriers and reagents was motivated by the need to ensure safety of application in the production of food-grade products. The CA and AI900 both recorded impressive immobilization yields of 82 and 62%, respectively, while the DMM recorded 47%. Enzyme immobilizations on CA, AI900 and DMM showed activity recoveries of 23, 27, and 17%, respectively. The CA, AI900 immobilized and the free enzymes recorded their highest scFOS yields of 59, 53, and 61%, respectively. The AI900 immobilized enzyme produced a consistent scFOS yield and composition for 12 batch cycles but for the CA immobilized enzyme, only 6 batch cycles gave a consistent scFOS yield. In its first record of application in scFOS production, the AI900 anion exchange resin exhibited potential as an adequate carrier for industrial application with possible savings on cost of immobilization and reduced technical difficulty. K E Y W O R D S adsorption, entrapment, fructooligosaccharides, frustosyltransferase, immobilization, ion exchange

Comparison of immobilized and free enzyme systems in industrial production of short‐chain fructooligosaccharides from sucrose using a techno‐economic approach

“…A summary of the mass and energy balances for the investigated scenarios is displayed in Table . The results obtained from experimental work recorded scFOS yields of 61%, 59% and 51% (w/w total sugars ) for FE, CAIE, and AIE respectively; the observed yields could be maintained for the CAIE and AIE methods for 6 and 12 recycles of reuse, respectively, before decreases were observed. Based on these results, significant reductions of 33% and 86% in the demand for the high‐cost enzyme biocatalyst for the CAIE and AIE systems, respectively, were achieved.…”

“…The selected process for this study was the two‐stage submerged fermentation, which is the most commonly applied system in commercial scFOS production . Based on the selected process descriptions, process simulations were built in Aspen Plus® V8.8 software (Aspen Technology Inc., USA), using experimental data from a previous study and data from the other published works …”

“…Results from previous work obtained an scFOS yield of 61% (w/w total sugars ) using the free enzyme and was therefore adopted for the industrial simulation as well. The 60% (w/v) sucrose solution is prepared by mixing sucrose at a feed rate of 442 kg h −1 with 295 kg h −1 of water in a blending tank (MIXER in Fig.…”

“…However, scFOS yields are usually lower than the free enzyme counterpart, due to the loss of activity from mass transfer limitation and inherent loss of activity during the enzyme immobilization process . Another possible shortcoming is that, unlike most conventional heterogenous catalysts, the immobilized biocatalyst in many cases experiences a substantial loss in activity after a limited number of reuses, typically between 7 and 15 cycles of reuse . As a result of the technical and functional advantages of the immobilized over the free enzyme reported in literature, many authors have proposed that the industrial application of an immobilized β ‐fructofuranosidase in scFOS production may offer economic advantages over the free enzyme …”

“…In this study, the production of scFOS from sucrose by transfructosylating reaction of the β ‐fructofuranosidase enzyme was considered . Three different enzyme processes (free, calcium alginate, and Amberlite IRA 900 immobilized enzymes) were compared for scFOS production to investigate the economic prospects of the immobilized enzyme in comparison with the free enzyme counterpart.…”