Magnetically separable spent coffee grounds as a potential novel support for the covalent immobilization of β-glucosidase for cellobiose hydrolysis

IF 2.8 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of chemical technology and biotechnology Pub Date : 2024-07-01 DOI:10.1002/jctb.7700

Shaifali Bhardwaj, Ankit Mishra, Debashish Ghosh, Anil Kumar Sinha

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引用次数: 0

Abstract

BACKGROUND

The industrial-scale application of enzymes faces obstacles due to elevated costs and difficulties in stability and reuse. In this study, magnetic spent coffee grounds, an ecotoxic waste, have been utilized successfully for the first time to immobilize β-glucosidase to overcome these challenges.

RESULTS

The spent coffee grounds were magnetized and amine-functionalized, followed by characterization using various techniques. Under optimized conditions, forming an imine bond between the functionalized support and β-glucosidase resulted in a 62% immobilization yield (92.81 mg g⁻¹ enzyme loading) and 12.5 U mg⁻¹ activity after immobilization. A relatively small kinetic change was observed in the K_m value (902 to 946 μmol L⁻¹) after immobilization, suggesting minimal hindrance by AMSCG₃ on substrate access or product release. Moreover, Glu@AMSCG₃ showed exceptional stability (>90% residual activity) within a pH range of 3 to 6 after 2 h of incubation at 25 °C. A residual activity of 87.94% was maintained even at 80 °C and pH 5 after 2 h of incubation compared to the free β-glucosidase, which showed only 6.5% residual activity at the same temperature. When cellobiose was hydrolyzed using Glu@AMSCG₃ under optimum conditions, 91.33% cellobiose conversion was achieved initially, and over 79% conversion was maintained for 10 reusability cycles.

CONCLUSION

The improved stability of β-glucosidase after covalent immobilization on amine-modified magnetically separable spent coffee grounds indicates their potential as a support matrix for application in enzyme immobilization. © 2024 Society of Chemical Industry (SCI).

查看原文本刊更多论文

将磁性可分离的废咖啡渣作为共价固定β-葡萄糖苷酶以水解纤维生物糖的潜在新型支持物

背景酶的工业规模应用面临着成本高、稳定性和再利用困难等障碍。本研究首次成功地利用磁性废咖啡渣（一种具有生态毒性的废物）来固定 β-葡萄糖苷酶，从而克服了这些挑战。在优化的条件下，功能化支撑物与 β-葡萄糖苷酶之间形成亚胺键后，固定化率为 62%（92.81 毫克/克-1 的酶负荷），固定化后的活性为 12.5 U 毫克/克-1。固定化后，Km 值（902 至 946 μmol L-1）的动力学变化相对较小，表明 AMSCG3 对底物获取或产物释放的阻碍极小。此外，Glu@AMSCG3 在 25 °C 下培养 2 小时后，在 pH 值为 3 到 6 的范围内表现出了极高的稳定性（90% 的残余活性）。即使在 80 °C、pH 值为 5 的条件下，经过 2 小时的培养，也能保持 87.94% 的残余活性，而游离的 β-葡萄糖苷酶在相同温度下的残余活性仅为 6.5%。在最佳条件下使用 Glu@AMSCG3 对纤维生物糖进行水解，最初纤维生物糖的转化率为 91.33%，在 10 个重复使用周期中转化率保持在 79% 以上。© 2024 化学工业学会（SCI）。

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来源期刊

Journal of chemical technology and biotechnology 工程技术-工程：化工

CiteScore

7.00

自引率

5.90%

发文量

268

审稿时长

1.7 months

期刊介绍： Journal of Chemical Technology and Biotechnology(JCTB) is an international, inter-disciplinary peer-reviewed journal concerned with the application of scientific discoveries and advancements in chemical and biological technology that aim towards economically and environmentally sustainable industrial processes.