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Plant aquaporin reconstituted proteoliposomes as nanosystem for resveratrol encapsulation
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  • Lucia Yepes-Molina,
  • Jose Teruel,
  • Urban Johanson,
  • Micaela Carvajal
Lucia Yepes-Molina
CEBAS
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Jose Teruel
University of Murcia
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Urban Johanson
Lund University
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Micaela Carvajal
CEBAS

Corresponding Author:mcarvaja@cebas.csic.es

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Abstract

Aquaporins (AQPs) are intrinsic membrane proteins responsible for facilitating water transport across biological membranes. AQPs found in plant membrane vesicles (MV) have been related to the functionality and stability of the vesicles. In this study, we focused on AQPs obtained from Brassica oleracea var. L. italica (broccoli) by the great potential for different biotechnological applications. To gain further insight into the role of AQPs in MV and advance the biotechnological applications of AQPs, we describe the heterologous overexpression of two broccoli AQPs (BoPIP1;2 and BoPIP2;2) in Pichia pastoris, resulting in the purification of both AQPs with high yield (0.14 and 0.99 mg per gram cells for BoPIP1;2 and BoPIP2;2, respectively). We reconstituted purified AQPs in liposomes to study their functionality, showing no changes in size compared to liposomes. BoPIP2;2 facilitated water transport, which was preserved for seven days at 4oC and 25ºC but not at 37oC, whereas BoPIP1;2 did not enhance water transport across the proteoliposome membrane. Additionally, BoPIP2;2 was incorporated into liposomes to encapsulate a resveratrol extract in proteoliposome vesicles, resulting in increased entrapment efficiency compared to conventional liposomes. Molecular docking identified potential binding sites for resveratrol in PIP2s, highlighting the role of AQPs in the improved entrapment efficiency of resveratrol. Moreover, a modelling study was conducted, demonstrating interactions between a plant AQP and human integrin, which may be a benefit to increase contact and internalization by the human target cells. Thus, our results suggest that AQPs-based alternative encapsulation systems can be used in specifically target biotechnological applications.
11 Aug 2023Submitted to Biotechnology Journal
14 Aug 2023Submission Checks Completed
14 Aug 2023Assigned to Editor
17 Aug 2023Reviewer(s) Assigned
26 Sep 2023Review(s) Completed, Editorial Evaluation Pending
26 Sep 2023Editorial Decision: Revise Major
28 Sep 20231st Revision Received
28 Sep 2023Submission Checks Completed
28 Sep 2023Assigned to Editor
28 Sep 2023Reviewer(s) Assigned
08 Oct 2023Review(s) Completed, Editorial Evaluation Pending
18 Oct 2023Editorial Decision: Revise Major