Lovastatin production by an oleaginous fungus, Aspergillus terreus KPR12 using sago processing wastewater (SWW).

BACKGROUND: Lovastatin is one of the first statins to be extensively used for its cholesterol-lowering ability. It is commercially produced by fermentation. Species belonging to the genus Aspergillus are well-studied fungi that have been widely used for lovastatin production. In the present study, we produced lovastatin from sago processing wastewater (SWW) under submerged fermentation using oleaginous fungal strains, A. terreus KPR12 and A. caespitosus ASEF14. RESULTS: The intra- and extracellular concentrations of lovastatin produced by A. terreus KPR12 and A. caespitosus ASEF14 were lactonized. Because A. caespitosus ASEF14 produced a negligible amount of lovastatin, further kinetics of lovastatin production in SWW was studied using the KPR12 strain for 9 days. Lovastatin concentrations in the intra- and extracellular fractions of the A. terreus KPR12 cultured in a synthetic medium (SM) were 117.93 and 883.28 mg L-1, respectively. However, these concentrations in SWW were 142.23 and 429.98 mg L-1, respectively. The yeast growth inhibition bioassay confirmed the antifungal property of fungal extracts. A. terreus KPR12 showed a higher inhibition zone of 14 mm than the ASEF14 strain. The two-way analysis of variance (ANOVA; p < 0.01) showed significant differences in the localization pattern, fungal strains, growth medium, and their respective interactions. The lovastatin yield coefficient values were 0.153 g g-1 on biomass (YLOV/X) and 0.043 g g-1 on the substrate, starch (YLOV/S). The pollutant level of treated SWW exhibited a reduction in total solids (TS, 59%), total dissolved solids (TDS, 68%), biological oxygen demand (BOD, 79.5%), chemical oxygen demand (COD, 57.1%), phosphate (88%), cyanide (65.4%), and void of nutrients such as nitrate (100%), and ammonia (100%). CONCLUSION: The starch-rich wastewater serves as a suitable medium for A. terreus KPR12 for the production of lovastatin. It simultaneously decontaminates the sago processing wastewater, enabling its reuse for irrigation/recreation.

Bioconversion of sago processing wastewater into biodiesel: Optimization of lipid production by an oleaginous yeast, Candida tropicalis ASY2 and its transesterification process using response surface methodology.

BACKGROUND: Biodiesel is an eco-friendly and renewable energy source and a valuable substitute for petro-diesel. Sago processing wastewater (SWW), a by-product of the cassava processing industry, has starch content ranging from 4 to 7 g L-1 and serves as an outstanding source for producing microbial lipids by the oleaginous microorganisms. In the present study, Candida tropicalis ASY2 was employed to optimize single-cell oil (SCO) production using SWW and subsequent transesterification by response surface methodology. Variables such as starch content, yeast extract, airflow rate, pH, and temperature significantly influenced lipid production in a preliminary study. The lipid production was scaled up to 5 L capacity airlift bioreactor and its optimization was done by response surface methodology. The dried yeast biomass obtained under optimized conditions from 5 L bioreactor was subjected to a direct transesterification process. Biomass: methanol ratio, catalyst concentration, and time were the variables used to attain higher FAME yield in the transesterification optimization process. RESULTS: Under optimized conditions, the highest lipid yield of 2.68 g L-1 was obtained with 15.33 g L-1 of starch content, 0.5 g L-1 of yeast extract, and 5.992 L min-1 of airflow rate in a bioreactor. The optimized direct transesterification process yielded a higher FAME yield of 86.56% at 1:20 biomass: methanol ratio, 0.4 M catalyst concentration, and a time of 6.85 h. CONCLUSIONS: Thus, this optimized process rendered the microbial lipids derived from C. tropicalis ASY2 as potentially alternative oil substitutes for sustainable biodiesel production to meet the rising energy demands.

Aspergillus caespitosus ASEF14, an oleaginous fungus as a potential candidate for biodiesel production using sago processing wastewater (SWW).

BACKGROUND: Oleaginous microorganisms are sustainable alternatives for the production of biodiesel. Among them, oleaginous fungi are known for their rapid growth, short life cycles, no light requirement, easy scalability, and the ability to grow in cheap organic resources. Among all the sources used for biodiesel production, industrial wastewater streams have been least explored. We used oleaginous fungi to decontaminate sago processing wastewater and produce biodiesel. RESULTS: Among the 15 isolates screened for lipid production and starch utilization using the Nile red staining assay and amylase plate screening, three isolates accumulated > 20% (w/w) of their dry cell mass as lipids. The isolate ASEF14 exhibited the highest lipid accumulation (> 40%) and was identified as Aspergillus caespitosus based on the 28S rRNA gene sequencing. The maximum lipid content of 54.4% in synthetic medium (SM) and 37.2% in sago processing wastewater (SWW) was produced by the strain. The Fourier-transform infrared (FTIR) spectroscopy of the fungal oil revealed the presence of functional peaks corresponding to major lipids. Principal component analysis (PCA) of the FTIR data revealed major changes in the fatty acid composition during the transition from the growth phase (Days 1-3) to the lipid accumulation phase (Days 4-7). The fatty acid methyl esters (FAME) analysis of fungal oil from SWW contained 43.82% and 9.62% of saturated and monounsaturated fatty acids, respectively. The composition and percentage of individual FAME derived from SWW were different from SM, indicating the effect of nutrient and fermentation time. The fuel attributes of the SM- and SWW-grown fungal biodiesel (kinematic viscosity, iodine value, cetane number, cloud and pour point, linolenic acid content, FA > 4 double bonds) met international (ASTM D6751, EN 14214) and national (IS 15607) biodiesel standards. In addition to biodiesel production, the strain removed various contaminants such as total solids (TS), total suspended solids (TSS), total dissolved solids (TDS), dissolved oxygen (DO), chemical oxygen demand (COD), biological oxygen demand (BOD), total nitrogen (TN), total phosphorus (TP), and cyanide up to 58.6%, 53.0%, 35.2%, 94.5%, 89.3%, 91.3%, 74.0%, 47.0%, and 53.84%, respectively, from SWW. CONCLUSION: These findings suggested that A. caespitosus ASEF14 is a potential candidate with high lipid accumulating ability (37.27%), capable of using SWW as the primary growth medium. The medium and incubation time alter the FAME profile of this fungus. The physical properties of fungal oil were in accordance with the biodiesel standards. Moreover, it decontaminated SWW by reducing several polluting nutrients and toxicants. The fungal biodiesel produced by this cost-effective method could serve as an alternate path to meet global energy demand.

Simultaneous lipid production for biodiesel feedstock and decontamination of sago processing wastewater using Candida tropicalis ASY2.

BACKGROUND: Without sufficient alternatives to crude oil, as demand continues to rise, the global economy will undergo a drastic decline as oil prices explode. Dependence on crude oil and growing environmental impairment must eventually be overcome by creating a sustainable and profitable alternative based on renewable and accessible feedstock. One of the promising solutions for the current and near-future is the substitution of fossil fuels with sustainable liquid feedstock for biofuel production. Among the different renewable liquid feedstock's studied, wastewater is the least explored one for biodiesel production. Sago wastewater is the byproduct of the cassava processing industry and has starch content ranging from 4 to 7%. The present investigation was aimed to produce microbial lipids from oleaginous yeast, Candida tropicalis ASY2 for use as biodiesel feedstock and simultaneously decontaminate the sago processing wastewater for reuse. Initial screening of oleaginous yeast to find an efficient amylolytic with maximum lipid productivity resulted in a potent oleaginous yeast strain, C. tropicalis ASY2, that utilizes SWW as a substrate. Shake flask experiments are conducted over a fermentation time of 240 h to determine a suitable fatty acid composition using GC-FID for biodiesel production with simultaneous removal of SWW pollutants using ASY2. RESULTS: The maximum biomass of 0.021 g L-1 h-1 and lipid productivity of 0.010 g L-1 h-1 was recorded in SWW with lipid content of 49%. The yeast strain degraded cyanide in SWW (79%) and also removed chemical oxygen demand (COD), biological oxygen demand (BOD), nitrate (NO3), ammoniacal (NH4), and phosphate (PO4) ions (84%, 92%, 100%, 98%, and 85%, respectively). GC-FID analysis of fatty acid methyl esters (FAME) revealed high oleic acid content (41.33%), which is one of the primary fatty acids for biodiesel production. CONCLUSIONS: It is evident that the present study provides an innovative and ecologically sustainable technology that generates valuable fuel, biodiesel using SWW as a substrate and decontaminates for reuse.