IBB-Institute for Biotechnology and Bioengineering, Centre for Biological and Chemical Engineering, Universidade Técnica de Lisboa, Av. Rovisco Pais, Lisbon, Portugal.
Appl Biochem Biotechnol. 2012 Nov;168(5):1197-211. doi: 10.1007/s12010-012-9850-1. Epub 2012 Sep 2.
The synthesis of galacto-oligosaccharides (GOS) by β-galactosidase immobilized in both polyvinyl alcohol (PVA) lenses and sol-gel carriers was studied and compared with the performance of the free enzyme. PVA-immobilized β-galactosidase retained 95 % of the initial activity after seven repeated uses and retained 51 % of the initial activity after 3 months of storage, while sol-gel-immobilized β-galactosidase only retained 39 % of the initial activity under storage. Lactose conversion takes place at a higher rate in the PVA-immobilized β-galactosidase, while the lowest rate of lactose conversion was noticed with immobilized β-galactosidase in sol-gel. Continuous production of GOS from either lactose or whey, with PVA-immobilized β-galactosidase, was performed in a packed-bed reactor. A maximum GOS production of 30 % of total sugars was attained for a 40-% lactose feed solution with a feed rate of 10.8 ml/h, at pH 4.5 and 40 °C, corresponding to a productivity of 117 g/l h. The maximum GOS productivity of 344 g/l h was obtained at a flow rate of 28.7 ml/h. 3-OS and 4-OS were the major types of GOS formed. Conversion of whey in continuous mode resulted in GOS production of 15 % of total sugars and formation of 45 % 3-OS, 40 % 4-OS, and 15 % 5-OS.
固定化β-半乳糖苷酶在聚乙烯醇(PVA)镜片和溶胶-凝胶载体中的半乳糖低聚糖(GOS)合成及其性能与游离酶进行了研究和比较。PVA 固定化β-半乳糖苷酶经 7 次重复使用后保留了初始活性的 95%,在储存 3 个月后保留了初始活性的 51%,而溶胶-凝胶固定化β-半乳糖苷酶在储存条件下仅保留了初始活性的 39%。在 PVA 固定化β-半乳糖苷酶中,乳糖转化的速率较高,而在溶胶-凝胶中固定化β-半乳糖苷酶的乳糖转化速率最低。使用 PVA 固定化β-半乳糖苷酶在填充床反应器中连续从乳糖或乳清生产 GOS。在 pH 值为 4.5 和 40°C、乳糖进料浓度为 40%、进料速率为 10.8 ml/h 的条件下,可获得 30%总糖的最大 GOS 产量,对应的产率为 117 g/l·h。在流速为 28.7 ml/h 的条件下,可获得最大的 GOS 产率 344 g/l·h。3-OS 和 4-OS 是形成 GOS 的主要类型。连续模式下乳清的转化导致 GOS 的总糖产量达到 15%,形成 45%的 3-OS、40%的 4-OS 和 15%的 5-OS。