Diverting the Use of Hand-Operated Tablet Press Machines to Bioassays: A Novel Protocol to Test 'Waste' Insoluble Shell Matrices

Methods Protoc. 2024 Apr 1;7(2):30. doi: 10.3390/mps7020030.

Abstract

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.

Keywords: bioactive factors; hand-operated tablet press machine; insoluble matrix; recycling; shell extract.

Grants and funding

CLT is the recipient of a PhD fellowship under joint-supervision between Université de Bourgogne (uB) and Universtà di Bologna Alma Mater Studiorum (UniBo). The research leading to these results has been conceived under the International PhD Program “Innovative Technologies and Sustainable Use of Mediterranean Sea Fishery and Biological Resources” (www.FishMed-PhD.org). This study represents partial fulfilment of the requirements for the Ph.D. thesis of CLT at the FishMed PhD Program (University of Bologna, Italy). In addition, CLT was supported by a one-off grant from AFFDU (Association des Femmes Françaises Diplômées de l’Université) in 2022 and an award from the Société Française de Biologie des Tissus Minéralisés (SFBTM) and from the Fondation Arthritis, in May 2023. Other supports to FM include the following: (1) the MAELSTROM project (les MAtricEs coquiLlières bacTéRicides au secOurs des géoMatériaux) from TELLUS-INTERRVIE program (CNRS, 2022); (2) the PRELUDE project (Peptides antimicRobiens dEs coquiLles de mollUsques D’intérêt Economique) financed by Observatoire des Sciences de l’Univers (OSU) Theta, Besançon, in 2023; (3) the MAELSTROM-bis project, financed by the TELLUS-INTERRVIE program from CNRS, in 2024.