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杂交生物微球的研制:纤维素暴露功能化融合蛋白。

Development of hybrid biomicroparticles: cellulose exposing functionalized fusion proteins.

机构信息

Department of Molecular Biotechnology, Faculty of Chemistry, University of Gdansk, Gdansk, 80-308, Poland.

BioVentures Institute Ltd, Poznan, 60-141, Poland.

出版信息

Microb Cell Fact. 2024 Mar 14;23(1):81. doi: 10.1186/s12934-024-02344-x.

Abstract

BACKGROUND

One of the leading current trends in technology is the miniaturization of devices to the microscale and nanoscale. The highly advanced approaches are based on biological systems, subjected to bioengineering using chemical, enzymatic and recombinant methods. Here we have utilised the biological affinity towards cellulose of the cellulose binding domain (CBD) fused with recombinant proteins.

RESULTS

Here we focused on fusions with 'artificial', concatemeric proteins with preprogrammed functions, constructed using DNA FACE™ technology. Such CBD fusions can be efficiently attached to micro-/nanocellulose to form functional, hybrid bionanoparticles. Microcellulose (MCC) particles were generated by a novel approach to enzymatic hydrolysis using Aspergillus sp. cellulase. The interaction between the constructs components - MCC, CBD and fused concatemeric proteins - was evaluated. Obtaining of hybrid biomicroparticles of a natural cellulose biocarrier with proteins with therapeutic properties, fused with CBD, was confirmed. Further, biological tests on the hybrid bioMCC particles confirmed the lack of their cytotoxicity on 46BR.1 N fibroblasts and human adipose derived stem cells (ASCs). The XTT analysis showed a slight inhibition of the proliferation of 46BR.1 N fibroblasts and ACSs cells stimulated with the hybrid biomicroparticles. However, in both cases no changes in the morphology of the examined cells after incubation with the hybrid biomicroparticles' MCC were detected.

CONCLUSIONS

Microcellulose display with recombinant proteins involves utilizing cellulose, a natural polymer found in plants, as a platform for presenting or displaying proteins. This approach harnesses the structural properties of cellulose to express or exhibit various recombinant proteins on its surface. It offers a novel method for protein expression, presentation, or immobilization, enabling various applications in biotechnology, biomedicine, and other fields. Microcellulose shows promise in biomedical fields for wound healing materials, drug delivery systems, tissue engineering scaffolds, and as a component in bio-sensors due to its biocompatibility and structural properties.

摘要

背景

目前,技术领域的一个主要趋势是将设备小型化到微尺度和纳米尺度。这些高度先进的方法基于生物系统,并通过化学、酶和重组方法进行生物工程化处理。在这里,我们利用纤维素结合域(CBD)与重组蛋白融合后的生物对纤维素的亲和力。

结果

在这里,我们专注于与使用 DNA FACE™ 技术构建的具有预编程功能的“人工”串联蛋白融合。这种 CBD 融合可以有效地附着在微/纳米纤维素上,形成功能性的杂交生物纳米颗粒。使用 Aspergillus sp. 纤维素酶通过一种新颖的酶解方法生成微纤维素 (MCC) 颗粒。评估了构建物组件 - MCC、CBD 和融合的串联蛋白 - 之间的相互作用。证实了具有治疗特性的融合蛋白与天然纤维素生物载体微纤维素的杂交生物复合颗粒的获得。此外,对杂交生物 MCC 颗粒的生物测试证实,它们对 46BR.1 N 成纤维细胞和人脂肪来源干细胞(ASCs)没有细胞毒性。XTT 分析显示,用杂交生物复合颗粒刺激的 46BR.1 N 成纤维细胞和 ACSs 细胞的增殖略有抑制。然而,在两种情况下,在用杂交生物复合颗粒的 MCC 孵育后,都没有检测到被检细胞形态的变化。

结论

带有重组蛋白的微纤维素展示涉及利用纤维素,即植物中发现的天然聚合物,作为展示或呈现蛋白质的平台。这种方法利用纤维素的结构特性,在其表面表达或展示各种重组蛋白。它为蛋白质表达、呈现或固定化提供了一种新方法,在生物技术、生物医学和其他领域有各种应用。微纤维素由于其生物相容性和结构特性,在生物医学领域具有很大的应用潜力,可用于伤口愈合材料、药物输送系统、组织工程支架以及生物传感器的组成部分。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a18/10938831/eabe166996e0/12934_2024_2344_Fig1_HTML.jpg

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