Abstract
Advances in biochemical and molecular manipulation have led to increased biomass productivity and oil accumulation in the microalgae C. reinhardtii. However, scalable processes for the recovery of oil and other valuable biomolecules, such as protein, from C. reinhardtii are scarce. The use of aqueous enzymatic extraction, a non-solvent and environmentally friendly bioproduct recovery method, provides an opportunity to design an integrated process for oil and protein fractionation to reduce bioenergy and bioproducts costs. Based on the mechanistic understanding of biomolecule distribution and compartmentalization, an aqueous enzymatic treatment for the release of internally stored lipid bodies was designed. Application of a C. reinhardtii-produced protease, autolysin, for lysis of the microalgae cell wall was followed by a secondary treatment with trypsin for chloroplast disruption and lipid body release. Protein recovery after the primary treatment with autolysin indicated a 50.1 ± 4.2% release of total soluble protein and localization of lipid bodies still in the chloroplast. The development of a secondary enzyme treatment (trypsin) for chloroplast and lipid body lysis demonstrated a high percent of remaining lipids (73 ± 7%) released into the supernatant. The results indicate that the application of an enzymatic treatment scheme for protein and oil recovery is a promising alternative to traditional extraction processes.
Highlights
The need to replace fossil fuels, one of the main causes of Green House Gas Emissions (GHGE) (Schenk et al 2008) has driven new research focused on exploring renewable energy sources
Proteins are more vulnerable to temperature and shear induced degradation
Impact of temperature and time on cell disruption and protein and lipid recovery Prior work by Soto-Sierra, Dixon (Soto-Sierra et al 2017) demonstrated that treating C. reinhardtii with autolysin for 4 h at 25°C was an effective method for cell lysis and resulted in ~ 20% protein release
Summary
The need to replace fossil fuels, one of the main causes of Green House Gas Emissions (GHGE) (Schenk et al 2008) has driven new research focused on exploring renewable energy sources. Microalgae cultivation has several advantages over other crops including a higher photosynthetic efficiency, higher biomass production, and faster growth rates (Mata et al 2010). Lipids from algae are rich in saturated and unsaturated fatty acids such as oleic (18:1), palmitic (16:0), stearic (18:0), and linoleic (18:2) acids (Meng et al 2009), making them ideal for fuel production and as high value food products. Some microalgae species, such as Chlamydomonas reinhardtii, are known for producing lipids and proteins with potential applications in the food and pharmaceutical industries. Even though there are several advantages regarding the utilization of C. reinhardtii cells as a protein and lipid source, there are still some challenges regarding its cultivation and extraction
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