Express Method for Isolation of Ready-to-Use 3D Chitin Scaffolds from Aplysina archeri (Aplysineidae: Verongiida) Demosponge

Abstract

Sponges are a valuable source of natural compounds and biomaterials for many biotechnological applications. Marine sponges belonging to the order Verongiida are known to contain both chitin and biologically active bromotyrosines. Aplysina archeri (Aplysineidae: Verongiida) is well known to contain bromotyrosines with relevant bioactivity against human and animal diseases. The aim of this study was to develop an express method for the production of naturally prefabricated 3D chitin and bromotyrosine-containing extracts simultaneously. This new method is based on microwave irradiation (MWI) together with stepwise treatment using 1% sodium hydroxide, 20% acetic acid, and 30% hydrogen peroxide. This approach, which takes up to 1 h, made it possible to isolate chitin from the tube-like skeleton of A. archeri and to demonstrate the presence of this biopolymer in this sponge for the first time. Additionally, this procedure does not deacetylate chitin to chitosan and enables the recovery of ready-to-use 3D chitin scaffolds without destruction of the unique tube-like fibrous interconnected structure of the isolated biomaterial. Furthermore, these mechanically stressed fibers still have the capacity for saturation with water, methylene blue dye, crude oil, and blood, which is necessary for the application of such renewable 3D chitinous centimeter-sized scaffolds in diverse technological and biomedical fields.

3D Chitin Scaffolds from the Marine Demosponge Aplysina archeri as a Support for Laccase Immobilization and Its Use in the Removal of Pharmaceuticals

“…Geyer GmbH & Co. KG, Renningen, Germany) at 24 • C for 6 h to remove residual calcium carbonates. It was then neutralized by rinsing with deionized water [26]. Deproteinization and demineralization of the sponge skeleton were repeated for 144 h to obtain a completely soft and pigment-free chitinous scaffold ( Figure 2) for further use as a support in enzyme immobilization.…”

“…Samples of the verongiid demosponge Aplysina archeri (Higgin, 1875) ( Figure 1) were collected at depths of 10-25 m by scuba divers around the Caribbean islands of Saint Vincent and Curaçao in May-June 2017, during the Pacotilles expedition. All permits required for the study were obtained during that expedition, which complied with all relevant regulations [26]. The reagents used for isolation of chitin sponge skeleton-sodium hydroxide, acetic acid, and hydrogen peroxide-were supplied by Merck (Darmstadt, Germany).…”

“…As shown, a very high immobilization yield of 91% was achieved, corresponding to the extraordinary amount of immobilized enzyme (1820 mg of laccase deposited per 1 g of chitinous scaffold). The high immobilization yield and large quantity of deposited enzyme may be explained by the exceptional sorption properties of A. archeri chitin [26] as well the presence of numerous functional groups typical for this aminopolysaccharide, mainly carbonyl and hydroxyl, that are capable of providing effective and stable enzyme binding [46]. However, it should be highlighted that although the interactions formed are stable, they are based mainly on hydrogen bonds and adsorption interactions.…”

“…Thus, demosponges of the order Dictioceratida (also known as commercial bath sponges) [7,8] represent a renewable source of proteinaceous spongin scaffolds which have recently been reported as effective in applications in extreme biomimetics [9][10][11][12], waste treatment [13][14][15][16][17], electrochemistry [18], and enzyme immobilization [19]. Meanwhile, marine demosponges of the order Verongiida have been recognized as a renewable source of uniquely pre-structured 3D chitinous scaffolds [20][21][22][23][24][25][26][27][28] which have found applications in tissue engineering [6,21,[29][30][31][32][33][34], drug release [35], the development of hybrid materials [36][37][38][39][40], and environmental science [41,42]. Chitin of invertebrate origin has previously been studied by researchers in protein immobilization as a matrix for the immobilization of enzymes [43][44][45][46][47][48], including ...…”

“…A selected air-dried fragment of Aplysina archeri sponge skeleton is rigid and represents the typical tube-like morphology of verongiids (A). Decellularization with subsequent demineralization of this construct [6,26] leads to isolation of the flexible chitinous scaffold (B-D), which reproduces the shape, form, and structural features of the original tubular sponge skeleton, with a length of up to 1.5 m.…”