Extraction and purification of violacein from Yarrowia lipolytica cells using aqueous solutions of surfactants

Abstract

BACKGROUND The demand for colorants from natural bio‐based sources is increasing. Violacein is a natural purple‐blue hydrophobic pigment with interesting bioactivity whose expression in genetically modified Yarrowia lipolytica production was successfully achieved. RESULTS In this work, several surfactants were tested in the extraction of violacein from Y. lipolytica cells, and the operational conditions were optimized to maximize the extraction yield. After the optimization, the purification of violacein using aqueous biphasic systems (ABS) composed of Tween 20 and cholinium‐based ionic liquids (ILs) was pursued. The ABS were characterized and applied in the separation of violacein from contaminant proteins, reaching a maximum selectivity of 155 with violacein being fully concentrated in the Tween 20‐rich phase, with 80% of the contaminant proteins present in the extract being removed. CONCLUSION This work led to a conceptual downstream process based on a first step of (solid–liquid) extraction and a second step addressing the separation of violacein from contaminant proteins using ABS. © 2019 Society of Chemical Industry

Recovery of bacterioruberin and proteins using aqueous solutions of surface-active compounds

“…The surfactants tested were pre-selected considering previous works that identified them to be efficient in the extraction of biomolecules, namely carotenoids. 37,38,46 Table S1 in the ESI† reports the ability of the surface-active compounds tested (30 in total) to extract bacterioruberin. Fig.…”

A plasmid system with tunable copy number

“…We chose to focus on Violacein (Fig. 5A), a purple pigment formed by the condensation of two tryptophan molecules 37 which is commonly used to demonstrate biosynthesis optimization 38 . The violacein biosynthetic pathway contains five enzymes, VioABCDE, each under the control of a different promoter and spanning approximately 8kb of DNA (Fig.…”

Yarrowia lipolyticachassis strains engineered to produce aromatic amino acids via the shikimate pathway

“…Our chassis strains can therefore be used as tools for producing a wide range of AAA derivatives by introducing heterologous pathways for biobased aromatic compounds. Such molecules like naringenin, resveratrol or even violacein have recently been successfully produced in Y. lipolytica, showing that the expression of these heterologous pathways is efficient in this yeast (Lv et al, 2019;Kholany et al, 2019;Palmer et al, 2020;S aez-S aez et al, 2020;Gu et al, 2020a). S aez-S aez and colleagues observed that glucose performed better than glycerol for the production of resveratrol (S aez-S aez et al, 2020).…”

“…Similarly, our TRP chassis strain resulted in PVA production in the absence of precursors supplementation, while violacein is usually generated using TRP‐supplemented media. For example, Wong and colleagues produced violacein in Y. lipolytica with a CSM medium containing TRP (Wong et al ., 2017 ), whereas Kholany and colleagues utilized a minimal or rich medium containing TRP (25 mg l −1 ) (Kholany et al ., 2019 ).…”