A new method to immobilize urease in silk fibroin membrane by unidirectional nanopore dehydration.

IF 2.5 3区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Meng Zhang, Hai-Yan Wang, Yu-Qing Zhang
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Abstract

The immobilization of free enzymes is crucial for enhancing their stability in different environments, enabling reusability, and expanding their applications. However, the development of a straightforward immobilization method that offers stability, high efficiency, biocompatibility, and modifiability remains a significant challenge. Silk fibroin (SF) is a good carrier for immobilized enzymes and drugs. Here, we employed urease as a model enzyme and utilized our developed technology called unidirectional nanopore dehydration (UND) to efficiently dehydrate a regenerated SF solution containing urease in a single step, resulting in the preparation of a highly functionalized SF membrane immobilizing urease (UI-SFM). The preparation process of UI-SFM is based on an all-water system, which is mild, green and able to efficiently and stably immobilize urease in the membranes, maintaining 92.7% and 82.8% relative enzyme activity after 30 days of storage in dry and hydrated states, respectively. Additionally, we performed additional post-treatments, including stretching and cross-linking with polyethylene glycol diglycidyl ether (PEGDE), to obtain two more robust immobilized urease membranes (UI-SFMs and UI-SFMc). The thermal and storage stability of these two membranes were significantly improved, and the recovery ratio of enzyme activity reached more than 90%. After 10 repetitions of the enzymatic reaction, the activity recovery of UI-SFMs and UI-SFMc remained at 92% and 88%, respectively. The results suggest that both UND-based and post-treatment-developed membranes exhibit excellent urease immobilization capabilities. Furthermore, the enzyme immobilization method offers a straightforward and versatile approach for efficient and stable enzyme immobilization, while its flexible modifiability caters to diverse application requirements.

通过单向纳米孔脱水将脲酶固定在蚕丝纤维素膜上的新方法。
游离酶的固定化对于提高其在不同环境中的稳定性、实现可重复使用性以及扩大其应用领域至关重要。然而,开发一种具有稳定性、高效性、生物兼容性和可修改性的直接固定方法仍然是一项重大挑战。蚕丝纤维素(SF)是固定化酶和药物的良好载体。在此,我们以脲酶为模型酶,利用我们开发的单向纳米孔脱水(UND)技术,对含有脲酶的再生蚕丝纤维素溶液进行高效脱水,从而制备出固定脲酶的高功能化蚕丝纤维素膜(UI-SFM)。UI-SFM 的制备过程基于全水体系,温和、绿色,能高效、稳定地将脲酶固定在膜中,在干燥和水合状态下分别保存 30 天后,酶的相对活性仍能保持 92.7% 和 82.8%。此外,我们还进行了额外的后处理,包括拉伸和与聚乙二醇二缩水甘油醚(PEGDE)交联,以获得两种更稳固的固定化脲酶膜(UI-SFMs 和 UI-SFMc)。这两种膜的热稳定性和贮存稳定性得到了显著提高,酶活性的回收率达到了 90% 以上。在重复 10 次酶反应后,UI-SFMs 和 UI-SFMc 的活性恢复率分别保持在 92% 和 88%。结果表明,基于 UND 和后处理技术开发的膜都具有出色的脲酶固定能力。此外,酶固定化方法为高效稳定地固定化酶提供了一种直接而通用的方法,同时其灵活的可修改性也能满足不同的应用要求。
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来源期刊
Biotechnology Progress
Biotechnology Progress 工程技术-生物工程与应用微生物
CiteScore
6.50
自引率
3.40%
发文量
83
审稿时长
4 months
期刊介绍: Biotechnology Progress , an official, bimonthly publication of the American Institute of Chemical Engineers and its technological community, the Society for Biological Engineering, features peer-reviewed research articles, reviews, and descriptions of emerging techniques for the development and design of new processes, products, and devices for the biotechnology, biopharmaceutical and bioprocess industries. Widespread interest includes application of biological and engineering principles in fields such as applied cellular physiology and metabolic engineering, biocatalysis and bioreactor design, bioseparations and downstream processing, cell culture and tissue engineering, biosensors and process control, bioinformatics and systems biology, biomaterials and artificial organs, stem cell biology and genetics, and plant biology and food science. Manuscripts concerning the design of related processes, products, or devices are also encouraged. Four types of manuscripts are printed in the Journal: Research Papers, Topical or Review Papers, Letters to the Editor, and R & D Notes.
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