Sync between leucine, biotin and citric acid to improve lipid production by Yarrowia lipolytica on crude glycerol-based media

One of the major environmental problems is the highly toxic agro-industrial waste called olive mill wastewater (OMW), deriving from olive oil production. On the other hand, the continuous development of the biological liquid fuel industry (biodiesel and bioethanol) makes it mandatory the process and exploitation of their main by-products, crude glycerol. This study dealt with the biotechnological conversions of biodiesel-derived crude glycerol with the use of the non-conventional yeast Yarrowia lipolytica in media that had been diluted with OMWs. OMWs, employed as simultaneous liquid medium and substrate, is a new trend recently appearing in Industrial Biotechnology, where value-added metabolites could be produced with simultaneous partial detoxification (i.e. decolorization and phenol removal) of the used residue. In the present study, diluted OMWs (containing 2.0 g/L of total phenolic compounds) blended with 70.0 g/L crude glycerol were employed as substrates. Production of value-added compounds by Y. lipolytica strain ACA-YC 5031 was studied in nitrogen-limited media favoring the production of secondary metabolites (i.e. citric acid, polyols, microbial lipids, polysaccharides). Batch-flask cultures were carried out and the impact of the addition of different NaCl concentrations (1.0%, 3.0%, 5.0% w/w) added upon the biochemical behavior of the strain was studied. Remarkable biomass production was observed in all trials, while in the “blank” experiment (no OMWs and no salt added), the metabolism was shifted toward the synthesis of polyols (Σpolyols = mannitol + arabitol + erythritol > 20 g/L and maximum total citric acid-Cit (sum of citric and isocitric acid) = 10.5 g/L). Addition of OMWs resulted in Citmax = 32.7 g/L, while Σpolyols concentration dropped to <15 g/L. Addition of salt in the OMW-based media slightly reduced the produced biomass, while Cit production drastically increased, reaching a final value of 54.0 g/L (conversion yield of Cit produced per unit of glycerol consumed = 0.82 g/g) in the trial with addition of 5.0% NaCl. Finally, significant color and phenols removal were observed, evaluating the yeast as a decontamination medium for the OMW and a great candidate for the production of value-added compounds.

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Valorization of Crude Glycerol, Residue Deriving from Biodiesel- Production Process, with the Use of Wild-type New Isolated Yarrowia lipolytica Strains: Production of Metabolites with Pharmaceutical and Biotechnological Interest

Current Pharmaceutical Biotechnology ◽

10.2174/1389201020666190211145215 ◽

2019 ◽

Vol 20 (10) ◽

pp. 881-894 ◽

Cited By ~ 4

Author(s):

Dimitris Sarris ◽

Zoe Sampani ◽

Anna Rapti ◽

Seraphim Papanikolaou

Keyword(s):

Citric Acid ◽

Yarrowia Lipolytica ◽

Crude Glycerol ◽

Fetal Brain ◽

Biodiesel Production ◽

Added Value ◽

Substrate Uptake ◽

Glycerol Concentration ◽

Dry Cell Weight ◽

Lipolytica Strain

Background & Objective:Crude glycerol (Glol), used as substrate for screening eleven natural Yarrowia lipolytica strains in shake-flask experiments. Aim of this study was to assess the ability of the screened strains to produce biomass (dry cell weight; X), lipid (L), citric acid (Cit), mannitol (Man), arabitol (Ara) and erythritol (Ery), compounds presenting pharmaceutical and biotechnological interest, in glycerol-based nitrogen-limited media, in which initial glycerol concentration had been adjusted to 40 g/L.Methods:Citric acid may find use in biomedical engineering (i.e. drug delivery, tissue engineering, bioimaging, orthopedics, medical device coating, wound dressings). Polyols are considered as compounds with non-cariogenic and less calorigenic properties as also with low insulin-mediated response. Microbial lipids containing polyunsaturated fatty acids (PUFA) are medically and dietetically important (selective pharmaceutical and anticancer properties, aid fetal brain development, the sight function of the eye, hormonal balance and the cardio-vascular system, prevent reasons leading to type-2 diabetes, present healing and anti-inflammatory effects).Results:All strains presented satisfactory microbial growth (Xmax=5.34-6.26 g/L) and almost complete substrate uptake. The principal metabolic product was citric acid (Citmax=8.5-31.7 g/L). Production of cellular lipid reached the values of 0.33-0.84 g/L. Polyols were also synthesized as strain dependent compounds (Manmax=2.8-6.1 g/L, Aramax ~2.0 g/L, Erymax= 0.5-3.8 g/L). The selected Y. lipolytica strain ACA-DC 5029 presented satisfactory growth along with synthesis of citric acid and polyols, thus, was further grown on media presenting an increased concentration of Glol~75 g/L. Biomass, lipid and citric acid production presented significant enhancement (Xmax=11.80 g/L, Lmax=1.26 g/L, Citmax=30.8 g/L), but conversion yield of citric acid produced per glycerol consumed was decreased compared to screening trials. Erythritol secretion (Erymax=15.6 g/L) was highly favored, suggesting a shift of yeast metabolism from citric acid accumulation towards erythritol production. Maximum endopolysaccharides (IPS) concentration was 4.04 g/L with yield in dry weight 34.2 % w/w.Conclusion:Y. lipolytica strain ACA-YC 5029 can be considered as a satisfactory candidate grown in high concentrations of crude glycerol to produce added-value compounds that interest pharmaceutical and biotechnology industries.

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