Department of Process Engineering, Stellenbosch University, Stellenbosch, South Africa.
Department of Microbiology, Stellenbosch University, Stellenbosch, South Africa.
Biotechnol Prog. 2019 May;35(3):e2797. doi: 10.1002/btpr.2797. Epub 2019 Mar 12.
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.
将β-呋喃果糖苷酶固定用于短链果寡糖（scFOS）合成具有提高生物催化剂效率的潜力。然而，载体和固定化技术的选择是实现这种效率的关键。在这项研究中，测试了海藻酸钠（CA）、Amberlite IRA 900（AI900）和 Dowex Marathon MSA（DMM）作为固定化来源于日本曲霉的新型工程β-呋喃果糖苷酶用于 scFOS 合成的载体。评估了几种固定化参数，以确定载体固定化酶的有效性。根据产生的 scFOS 的产率和可重复使用性来比较固定化生物催化剂的性能。载体和试剂的选择是为了确保在生产食品级产品中的应用安全性。CA 和 AI900 的固定化产率分别高达 82%和 62%，而 DMM 的固定化产率为 47%。CA、AI900 和 DMM 上的酶固定化的酶活回收率分别为 23%、27%和 17%。CA、AI900 固定化酶和游离酶的 scFOS 产率分别达到 59%、53%和 61%的最高值。AI900 固定化酶在 12 批循环中产生一致的 scFOS 产率和组成，但 CA 固定化酶仅在 6 批循环中产生一致的 scFOS 产率。AI900 阴离子交换树脂在 scFOS 生产中的首次应用记录表明，它作为工业应用的合适载体具有潜力，可能节省固定化成本和降低技术难度。
