Incorporation of Magnetic Nanoparticles into Protoplasts of Microalgae Haematococcus pluvialis: A Tool for Biotechnological Applications.

Maria G Savvidou,Angelo Ferraro,Dimitris Kekos,Evangelos Hristoforou,Fragiskos N Kolisis,Diomi Mamma

doi:10.3390/molecules25215068

Maria G Savvidou, Angelo Ferraro + Show 4 more

Open Access

https://doi.org/10.3390/molecules25215068

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Abstract

Intensive research on the use of magnetic nanoparticles for biotechnological applications of microalgae biomass guided the development of proper treatment to successfully incorporate them into these single-cell microorganisms. Protoplasts, as cells lacking a cell wall, are extensively used in plant/microalgae genetic manipulation as well as various biotechnological applications. In this work, a detailed study on the formation of protoplasts from Haematococcus pluvialis with the use of enzymatic and mechanical procedures was performed. The optimization of several parameters affecting the formation of protoplasmic cells and cell recovery was investigated. In the enzymatic treatment, a solution of cellulase was studied at different time points of incubation, whereas in the mechanical treatment, glass beads vortexing was used. Mechanical treatment gave better results in comparison to the enzymatic one. Concerning the cell recovery, after the protoplast formation, it was found to be similar in both methods used; cell viability was not investigated. To enhance the protoplast cell wall reconstruction, different “recovery media” with an organic source of carbon or nitrogen were used. Cell morphology during all treatments was evaluated by electron microscopy. The optimal conditions found for protoplast formation and cell reconstruction were successfully used to produce Haematococcus pluvialis cells with magnetic properties.

Highlights

Magnetic nanoparticles are a new trend in various scientific fields, such as drug delivery, DNA/RNA purification, improved magnetic resonance imaging (MRI), immobilization, food industry, medical diagnostics, cell harvesting, bioremediation, and others [1,2,3,4]
In order to accomplish our goal, the creation of transformable H. pluvialis microalgae, two different approaches for protoplast formation were used; the enzymatic, which is based on cellulase treatment, and the mechanical, which is based on glass bead vortexing
Biotechnological or genetic transformation of microalgae cells could be succeeded through a supports the normal regeneration and growth of microalgae

Summary

Introduction

Magnetic nanoparticles are a new trend in various scientific fields, such as drug delivery, DNA/RNA purification, improved magnetic resonance imaging (MRI), immobilization, food industry, medical diagnostics, cell harvesting, bioremediation, and others [1,2,3,4]. They are a class of particles consisting of a magnetic and a functional chemical component with a diameter varying from 1 to 100 nm and can be controlled by magnetic fields displaying superparamagnetism. Heart and eye health, and photoprotection are fields where H. pluvialis is an important implement [6].

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