将手动压片机转用于生物测定:测试 "废弃 "不溶性贝壳基质的新方案。

IF 2.3 Q3 BIOCHEMICAL RESEARCH METHODS Methods and Protocols Pub Date : 2024-04-01 DOI:10.3390/mps7020030
Camille Lutet-Toti, Marie Da Silva Feliciano, Nelly Debrosse, Jérôme Thomas, Laurent Plasseraud, Frédéric Marin
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引用次数: 0

摘要

为了使贝壳矿化,软体动物会分泌一种复杂的鸡尾酒蛋白质--统称为钙化贝壳基质--这种蛋白质一直被封闭在外骨骼中。如今,从贝壳中提取蛋白质已被视为生物活性物质的潜在来源,其中信号分子、杀菌剂或蛋白酶抑制剂为应用科学、健康和水产养殖提供了最切实的前景。然而,测试贝壳提取物活性的一个技术障碍在于其高度不溶性。在本文中,我们提出了一种规避这一障碍的方案。在对贝壳蛋白质进行提取并生产出两种有机组分(一种可溶,一种不溶)后,我们使用手动压片机生产出由 100% 不溶性贝壳基质组成的校准良好的片剂。对片剂质量的傅立叶变换红外光谱监测表明,压片机使用的压力不会损害不溶性提取物的分子特性。生产出的药片可直接进行不同的生物检测,如培养皿中的杀菌抑菌区检测,如图所示。改用手动压片机为分析不溶性贝壳基质、发现新型生物活性成分开辟了新的前景。
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Diverting the Use of Hand-Operated Tablet Press Machines to Bioassays: A Novel Protocol to Test 'Waste' Insoluble Shell Matrices.
To mineralize their shells, molluscs secrete a complex cocktail of proteins-collectively defined as the calcifying shell matrix-that remains occluded in the exoskeleton. Nowadays, protein extracts from shells are recognized as a potential source of bioactive substances, among which signalling molecules, bactericides or protease inhibitors offer the most tangible perspectives in applied sciences, health, and aquaculture. However, one technical obstacle in testing the activity of shell extracts lies in their high insolubility. In this paper, we present a protocol that circumvents this impediment. After an adapted shell protein extraction and the production of two organic fractions-one soluble, one insoluble-we employ a hand-operated tablet press machine to generate well-calibrated tablets composed of 100% insoluble shell matrix. FT-IR monitoring of the quality of the tablets shows that the pressure used in the press machine does not impair the molecular properties of the insoluble extracts. The produced tablets can be directly tested in different biological assays, such as the bactericidal inhibition zone assay in Petri dish, as illustrated here. Diverting the use of the hand-operated tablet press opens new perspectives in the analysis of insoluble shell matrices, for discovering novel bioactive components.
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来源期刊
Methods and Protocols
Methods and Protocols Biochemistry, Genetics and Molecular Biology-Biochemistry, Genetics and Molecular Biology (miscellaneous)
CiteScore
3.60
自引率
0.00%
发文量
85
审稿时长
8 weeks
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