Laboratório de Biotecnologia de Alimentos, Universidade do Vale do Taquari - Univates, Lajeado, RS, Brazil; Programa de Pós-Graduação em Biotecnologia, Universidade do Vale do Taquari - Univates, Lajeado, RS, Brazil.
Laboratório de Biotecnologia de Alimentos, Universidade do Vale do Taquari - Univates, Lajeado, RS, Brazil.
Bioresour Technol. 2021 Apr;326:124747. doi: 10.1016/j.biortech.2021.124747. Epub 2021 Jan 23.
This study aimed to produce and characterize a recombinant Kluyveromyces sp. β-galactosidase fused to a cellulose-binding domain (CBD) for industrial application. In expression assays, the highest enzymatic activities occurred after 48 h induction on Escherichia coli C41(DE3) strain at 20 °C in Terrific Broth (TB) culture medium, using isopropyl β-d-1-thiogalactopyranoside (IPTG) 0.5 mM (108.77 U/mL) or lactose 5 g/L (93.10 U/mL) as inducers. Cultures at bioreactor scale indicated that higher product yield values in relation to biomass (2000 U/g) and productivity (0.72 U/mL.h) were obtained in culture media containing higher protein concentration. The recombinant enzyme showed high binding affinity to nanocellulose, reaching both immobilization yield and efficiency values of approximately 70% at pH 7.0 after 10 min reaction. The results of the present study pointed out a strategy for recombinant β-galactosidase-CBD production and immobilization, aiming toward the application in sustainable industrial processes using low-cost inputs.
本研究旨在生产和表征一种与纤维素结合域（CBD）融合的重组克鲁维酵母β-半乳糖苷酶，用于工业应用。在表达试验中，在大肠杆菌 C41(DE3)菌株上于 20°C 在 Terrific Broth (TB) 培养基中用异丙基-β-d-1-硫代半乳糖苷（IPTG）0.5mM（108.77U/mL）或乳糖 5g/L（93.10U/mL）诱导 48 小时后，酶活最高。在生物反应器规模的培养中，在含有较高蛋白质浓度的培养基中，与生物量（2000U/g）和生产率（0.72U/mL.h）相比，产物产率值更高。重组酶对纳米纤维素表现出高结合亲和力，在 pH 7.0 下反应 10 分钟后，固定化产率和效率值均接近 70%。本研究的结果提出了一种用于生产和固定化重组β-半乳糖苷酶-CBD 的策略，旨在为使用低成本投入的可持续工业过程应用提供参考。
