Covalent Immobilization of β-Glucosidase on Magnetic Spent Coffee Grounds Modified With PAMAM Dendrimers for Enhanced Glucose Tolerance

IF 3.1 3区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Biotechnology Journal Pub Date : 2025-06-16 DOI:10.1002/biot.70025

Shaifali Bhardwaj, Ekta Naik, Akansha Tripathi, Debashish Ghosh, Anil Kumar Sinha

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Abstract

The successful application of enzymes in industries encounters challenges related to high costs, stability, and reuse. In this study, β-glucosidase (BGL) was immobilized via a covalent method after the synthesis of different generations (G_0-2) of polyamidoamine (PAMAM) dendrimers modified magnetic spent coffee grounds (SCGs). With the increase in PAMAM generation, BGL immobilization (163.123 and 218.99 mg protein/g support in G₁ and G₂, respectively) on the support increased. Exceptional stability (> 80% residual activity) was observed for immobilized BGL at varying pH (3 to 6) and temperature (30°C to 80°C) after a 2 h incubation period. The higher generations of PAMAM dendrimers were more tolerant to glucose inhibition than the free enzyme. PAMAM G₁ and G₂ generations showed > 75% residual activity at 200 mM glucose concentration compared to free enzyme, with only 50% residual activity at 100 mM glucose concentration. After employing BGL@MSCG₃@G_0-2 for cellobiose hydrolysis, > 69% glucose yield for 10 cycles was maintained.

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PAMAM树状大分子修饰的磁咖啡渣共价固定β-葡萄糖苷酶增强葡萄糖耐量

酶在工业中的成功应用遇到了与高成本、稳定性和重用相关的挑战。在本研究中，合成不同代（G0-2）聚氨基胺（PAMAM）树状大分子修饰磁性废咖啡渣（SCGs）后，通过共价法固定化β-葡萄糖苷酶（BGL）。随着PAMAM代的增加，BGL固定化（G1和G2分别为163.123和218.99 mg蛋白/g）对支架的影响增加。卓越的稳定性(>；在不同的pH值（3 ~ 6）和温度（30 ~ 80℃）下，经过2小时的孵育，观察到固定化BGL的残余活性为80%。更高代的PAMAM树状大分子比游离酶更耐受葡萄糖抑制。PAMAM G1和G2代显示>；与游离酶相比，在200 mM葡萄糖浓度下剩余活性为75%，而在100 mM葡萄糖浓度下只有50%的剩余活性。采用BGL@MSCG3@G0-2进行纤维素二糖水解后，>；10个循环后葡萄糖产率维持在69%。

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来源期刊

Biotechnology Journal Biochemistry, Genetics and Molecular Biology-Molecular Medicine

CiteScore

8.90

自引率

2.10%

发文量

123

审稿时长

1.5 months

期刊介绍： Biotechnology Journal (2019 Journal Citation Reports: 3.543) is fully comprehensive in its scope and publishes strictly peer-reviewed papers covering novel aspects and methods in all areas of biotechnology. Some issues are devoted to a special topic, providing the latest information on the most crucial areas of research and technological advances. In addition to these special issues, the journal welcomes unsolicited submissions for primary research articles, such as Research Articles, Rapid Communications and Biotech Methods. BTJ also welcomes proposals of Review Articles - please send in a brief outline of the article and the senior author''s CV to the editorial office. BTJ promotes a special emphasis on: Systems Biotechnology Synthetic Biology and Metabolic Engineering Nanobiotechnology and Biomaterials Tissue engineering, Regenerative Medicine and Stem cells Gene Editing, Gene therapy and Immunotherapy Omics technologies Industrial Biotechnology, Biopharmaceuticals and Biocatalysis Bioprocess engineering and Downstream processing Plant Biotechnology Biosafety, Biotech Ethics, Science Communication Methods and Advances.