A multi-component reaction for covalent immobilization of lipases on amine-functionalized magnetic nanoparticles: production of biodiesel from waste cooking oil
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ABSTRACT:
SUBMITTER: Amini Y
PROVIDER: S-EPMC10991503 | biostudies-literature | 2022 May
REPOSITORIES: biostudies-literature
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Project description:Cooking oil waste leads to well-known environmental impacts and its bioremediation by lipase-based enzymatic activity can minimize the high cytotoxic potential. In addition, they are among the biocatalysts most commercialized worldwide due to the versatility of reactions and substrates. However, although lipases are able to process cooking oil wastes, the products generated from this process do not necessarily become less toxic. Thus, the aim of the current study is to analyze the bioremediation of lipase-catalyzed cooking oil wastes, as well as their effect on the cytotoxicity of both the oil and its waste before and after enzymatic treatment. Thus, assessed the post-frying modification in soybean oil and in its waste, which was caused by hydrolysis reaction catalyzed by commercial and home-made lipases. The presence of lipases in the extracts obtained from orange wastes was identified by zymography. The profile of the fatty acid esters formed after these reactions was detected and quantified through gas chromatography and fatty acids profile compared through multivariate statistical analyses. Finally, the soybean oil and its waste, with and without enzymatic treatment, were assessed for toxicity in cytotoxicity assays conducted in vitro using fibroblast cell culture. The soybean oil wastes treated with core and frit lipases through transesterification reaction were less toxic than the untreated oils, thus confirming that cooking oil wastes can be bioremediated using orange lipases.
| S-EPMC5657992 | biostudies-literature
Project description:Immobilized lipases were applied to the enzymatic conversion of oils from spent coffee ground into biodiesel. Two lipases were selected for the study because of their conformational behavior analysed by Molecular Dynamics (MD) simulations taking into account that immobilization conditions affect conformational behavior of the lipases and ultimately, their efficiency upon immobilization. The enzymatic synthesis of biodiesel was initially carried out on a model substrate (triolein) in order to select the most promising immobilized biocatalysts. The results indicate that oils can be converted quantitatively within hours. The role of the nature of the immobilization support emerged as a key factor affecting reaction rate, most probably because of partition and mass transfer barriers occurring with hydrophilic solid supports. Finally, oil from spent coffee ground was transformed into biodiesel with yields ranging from 55% to 72%. The synthesis is of particular interest in the perspective of developing sustainable processes for the production of bio-fuels from food wastes and renewable materials. The enzymatic synthesis of biodiesel is carried out under mild conditions, with stoichiometric amounts of substrates (oil and methanol) and the removal of free fatty acids is not required.
| S-EPMC4030945 | biostudies-literature
Project description:The world biodiesel production is increasing at a rapid rate. Despite its perceived safety for the environment, more detailed toxicity studies are mandatory, especially in the field of aquatic toxicology. While considerable attention has been paid to biodiesel combustion emissions, the toxicity of biodiesel in the aquatic environment has been poorly understood. In our study, we used an algae culture growth-inhibition test (OECD 201) for the comparison of the toxicity of B100 (pure biodiesel), produced by methanol transesterification of waste cooking oil (yellow grease), B0 (petroleum diesel fuel) and B20 (diesel-biodiesel blended of 20% biodiesel and 80% petroleum diesel fuel by volume). Two marine diatoms Attheya ussuriensis and Chaetoceros muelleri, the red algae Porphyridium purpureum and Raphidophyte Heterosigma akashiwo were employed as the aquatic test organisms. A sample of biodiesel from waste cooking oil without dilution with petroleum diesel (B100) showed the highest level of toxicity for the microalgae A. ussuriensis, C. muelleri and H. akashiwo, compared to hexane, methanol, petroleum diesel (B0) and diluted sample (B20). The acute EC50 in the growth-inhibition test (96 h exposure) of B100 for the four species was in the range of 3.75-23.95 g/L whereas the chronic toxicity EC50 (7d exposure) was in the range of 0.42-16.09 g/L.
| S-EPMC6317304 | biostudies-literature
Project description:Waste cooking oil (WCO) represents one of the most available renewable energy sources to produce solid alcohol biofuel (SABF) and biodiesel. The objective of this work is to conduct energy and cost balances for biodiesel and SABF generation from supernatant waste cooking oil (S-WCO) and bottom waste cooking oil (B-WCO) derived from catering facilities in Iran, respectively. The total energy input/output for SABF and biodiesel, respectively, were 31.738/37.180 MJ/L and 34.269/42.163 MJ/L, with the majority of the energy contribution being B-WCO/S-WCO (78.77/73.57 %), followed by machinery (19.76/18.30 %), and ethanol/methanol (0.69/7.47 %), in both cases. Renewable energy and non-renewable energy inputs for SABF and biodiesel were calculated as 25.071 MJ/L/6.667 MJ/L and 25.280 MJ/L/8.990 MJ/L of total energy input, respectively. Total cost and revenues for SABF vs. biodiesel production processes were 1.318 vs. 1.342 $/L and 5.620 vs. 2.481$/L, respectively. Gross return, net return and benefit to cost ratio values for SABF vs. biodiesel were 1.449 vs. 1.548 $/L, 1.060 vs. 1.157 $/L, and 1.827 vs. 1.876, respectively. Hence, considering the energy balances of the valorization process for WCO, the population growth in cities, and the need for inexpensive and plentiful sources of renewable energy, the production of SABF and biodiesel may offer a valuable solution in developing countries.
| S-EPMC11867234 | biostudies-literature
Project description:In this study, two highly thermotolerant and methanol-tolerant lipase-producing bacteria were isolated from cooking oil and they exhibited a high number of catalytic lipase activities recording 18.65 ± 0.68 U/mL and 13.14 ± 0.03 U/mL, respectively. Bacterial isolates were identified according to phenotypic and genotypic 16S rRNA characterization as Kocuria flava ASU5 (MT919305) and Bacillus circulans ASU11 (MT919306). Lipases produced from Kocuria flava ASU5 showed the highest methanol tolerance, recording 98.4% relative activity as well as exhibited high thermostability and alkaline stability. Under the optimum conditions obtained from 3D plots of response surface methodology design, the Kocuria flava ASU5 biocatalyst exhibited an 83.08% yield of biodiesel at optimized reaction variables of, 60 ○C, pH value 8 and 1:2 oil/alcohol molar ratios in the reaction mixture. As well as, the obtained results showed the interactions of temperature/methanol were significant effects, whereas this was not noted in the case of temperature/pH and pH/methanol interactions. The obtained amount of biodiesel from cooking oil was 83.08%, which was analyzed by a GC/Ms profile. The produced biodiesel was confirmed by Fourier-transform infrared spectroscopy (FTIR) approaches showing an absorption band at 1743 cm-1, which is recognized for its absorption in the carbonyl group (C=O) which is characteristic of ester absorption. The energy content generated from biodiesel synthesized was estimated as 12,628.5 kJ/mol. Consequently, Kocuria flava MT919305 may provide promising thermostable, methanol-tolerant lipases, which may improve the economic feasibility and biotechnology of enzyme biocatalysis in the synthesis of value-added green chemicals.
| S-EPMC8249636 | biostudies-literature
Project description:Waste cooking oil (WCO) is a readily available and cheap feedstock for biodiesel production. However, WCO contains high levels of free fatty acids (FFAs), which negatively impact the biodiesel yield if homogeneous catalysts are used. Heterogeneous solid acid catalysts are preferred for low-cost feedstocks because the catalysts are highly insensitive to high levels of FFA in the feedstock. Therefore, in the present study, we synthesized and evaluated different solid catalysts, pure β-zeolite, ZnO-β-zeolite, and SO42-/ZnO-β-zeolite for the production of biodiesel using WCO as feedstock. The synthesized catalysts were characterized by Fourier transform infrared spectroscopy (FTIR), pyridine-FTIR, N2 adsorption-desorption, X-ray diffraction, thermogravimetric analysis, and scanning electron microscopy, while the biodiesel product was analyzed using nuclear magnetic resonance (1H and 13C NMR) and gas chromatography-mass spectroscopy. The results revealed that the SO42-/ZnO-β-zeolite catalyst showed excellent catalytic performance for simultaneous transesterification and esterification of WCO, with a higher percentage conversion than the ZnO-β-zeolite and pure β-zeolite catalyst, due to the large pore size and high acidity. The SO42-/ZnO-β-zeolite catalyst exhibits 6.5 nm pore size, a total pore volume of 0.17 cm3/g, and high surface area of 250.26 m2/g. Experimental parameters such as catalyst loading, methanol:oil molar ratio, temperature, and reaction time were varied in order to establish the optimal parameters. The highest WCO conversion of 96.9% was obtained using the SO42-/ZnO-β-zeolite catalyst under an optimum reaction condition of 3.0 wt % catalyst loading, 200 °C reaction temperature, and 15:1 molar ratio of methanol to oil in 8 h reaction time. The WCO-derived biodiesel properties conform to the ASTM6751 standard specification. Our investigation of its kinetics revealed that the reaction follows a pseudo first-order kinetic model, with an activation energy (Ea) of 38.58 kJ/mol. Moreover, the stability and reusability of the catalysts were evaluated, and it was found that the SO42-/ZnO-β-zeolite catalyst exhibited good stability, giving a biodiesel conversion of over 80% after three synthesis cycles.
| S-EPMC10324085 | biostudies-literature
Project description:The present study focuses on the catalytic activity of cesium modified natural phosphate in biodiesel production from rapeseed oil and low free fatty acids (FFA) used in cooking oil. The catalyst was prepared by impregnation of cesium chloride (CsCl) on the natural phosphate followed by calcination up to 800 °C. The phosphate based catalyst was thermally, structurally, morphologically, and texturally characterized in order to determinate the relationship between its physicochemical properties and its catalytic activity. The chosen catalyst was demonstrated to be an active catalyst for the transesterification of rapeseed oil achieving a biodiesel yield of 99.55% under suitable reaction conditions: a methanol to oil molar ratio of 12 : 1, reaction temperature of 70 °C, catalyst amount of 4 wt% based on oil weight and reaction time of 6 h. Results from low FFA waste cooking oil transesterification indicated that a methyl esters yield of 99.52% could be obtained. Furthermore, results from esterification/transesterification of acidified rapeseed oil indicate that a yield of 93% may be obtained, thus giving rise to a potential application in 2nd generation biodiesel production from low acidic oils. Some important physicochemical properties of the obtained biodiesel were evaluated and compared with the EN14214 and ASTM D-6751 standards for biodiesel specifications.
| S-EPMC9057709 | biostudies-literature
Project description:BackgroundOleaginous yeasts are fast emerging as a possible feedstock for biodiesel production. Yarrowia lipolytica, a model oleaginous yeast is known to utilize a variety of hydrophobic substrates for lipid accumulation including waste cooking oil (WCO). Approaches to increase lipid content in this yeast include metabolic engineering which requires manipulation of multiple genes in the lipid biosynthesis pathway. A classical and cost-effective approach, namely, random chemical mutagenesis on the yeast can lead to increased production of biodiesel as is explored here.ResultsIn this study, chemical mutagenesis using the alkylating agent, N- methyl-N'-nitro-N-nitrosoguanidine (MNNG) as well as an additional treatment with cerulenin, a fatty acid synthase inhibitor generated 800 mutants of Y. lipolytica NCIM 3589 (761 MNNG treated and 39 MNNG + cerulenin treated). A three-stage screening using Sudan Black B plate technique, Nile red fluorimetry and total lipid extraction using solvent was performed, which enabled selection of ten high lipid yielding mutants. Time course studies of all the ten mutants were further undertaken in terms of biomass, lipid yield and lipid content to select three stable mutants (YlB6, YlC7 and YlE1) capable of growing and accumulating lipid on WCO, with lipid contents of 55, 60 and 67% as compared to 45% for the wild type. The mutants demonstrated increased volumetric lipid productivities (0.062, 0.044 and 0.041 g L-1 h-1) as compared to the wild type (0.033 g L-1 h-1). The fatty acid profile of the three mutants consisted of a high content of C16 and C18 saturated and monounsaturated fatty acids and was found to be suitable for biodiesel production. The fuel properties, namely, density, kinematic viscosity, total acid number, iodine value of the three mutants were evaluated and found to lie within the limits specified by internationally accepted standards. Additionally, it was noted that the mutants demonstrated better cetane numbers and higher heating values than the wild type strain.ConclusionThe chemical mutagenesis strategy adopted in this study resulted in the successful isolation of three stable high SCO yielding mutants. The mutants, namely, YlB6, YlC7 and YlE1 exhibited a 1.22, 1.33 and 1.49-fold increase in lipid contents when grown on 100 g L-1 waste cooking oil than the parental yeast strain. The fatty acid methyl ester (FAME) profiles of all the three mutants was determined to be suitable for biodiesel suggesting their potential applicability while simultaneously addressing the management of waste cooking oil.
| S-EPMC5655982 | biostudies-literature
Project description:BackgroundWe have recently developed a one-step, genetically encoded immobilization approach based on fusion of a target enzyme to the self-crystallizing protein Cry3Aa, followed by direct production and isolation of the fusion crystals from Bacillus thuringiensis. Using this approach, Bacillus subtilis lipase A was genetically fused to Cry3Aa to produce a Cry3Aa-lipA catalyst capable of the facile conversion of coconut oil into biodiesel over 10 reaction cycles. Here, we investigate the fusion of another lipase to Cry3Aa with the goal of producing a catalyst suitable for the conversion of waste cooking oil into biodiesel.ResultsGenetic fusion of the Proteus mirabilis lipase (PML) to Cry3Aa allowed for the production of immobilized lipase crystals (Cry3Aa-PML) directly in bacterial cells. The fusion resulted in the loss of PML activity, however, and so taking advantage of its genetically encoded immobilization, directed evolution was performed on Cry3Aa-PML directly in its immobilized state in vivo. This novel strategy allowed for the selection of an immobilized PML mutant with 4.3-fold higher catalytic efficiency and improved stability. The resulting improved Cry3Aa-PML catalyst could be used to catalyze the conversion of waste cooking oil into biodiesel for at least 15 cycles with minimal loss in conversion efficiency.ConclusionsThe genetically encoded nature of our Cry3Aa-fusion immobilization platform makes it possible to perform both directed evolution and screening of immobilized enzymes directly in vivo. This work is the first example of the use of directed evolution to optimize an enzyme in its immobilized state allowing for identification of a mutant that would unlikely have been identified from screening of its soluble form. We demonstrate that the resulting Cry3Aa-PML catalyst is suitable for the recyclable conversion of waste cooking oil into biodiesel.
| S-EPMC6598307 | biostudies-literature
Project description:Herein, we have designed and demonstrated a facile and effective platform for the covalent anchoring of a tetrameric hemoprotein, hemoglobin (Hb). The platform comprises of naphthyl substituted amine functionalized gel type hydrophobic ionic liquid (NpNH2-IL) through which the heme protein was covalently attached over a glassy carbon electrode (Hb-NpNH2-IL/GCE). UV-vis and FT-IR spectral results confirmed that the Hb on NpNH2-IL retains its native structure, even after being covalently immobilized on NpNH2-IL platform. The direct electron transfer of redox protein could be realized at Hb-NpNH2-IL/GCE modified electrode and a well resolved redox peak with a formal potential of -0.30 V and peak separation of 65 mV was observed. This is due to the covalent attachment of highly conducting NpNH2-IL to the Hb, which facilitates rapid shuttling of electrons between the redox site of protein and the electrode. Further, the fabricated biosensor favoured the electrochemical reduction of bromate in neutral pH with linearity ranging from 12 to 228 µM and 0.228 to 4.42 mM with a detection limit and sensitivities of 3 µM, 430.7 µA mM-1 cm-2 and 148.4 µA mM-1 cm-2 respectively. Notably, the fabricated biosensor showed good operational stability under static and dynamic conditions with high selectivity and reproducibility.
| S-EPMC6639313 | biostudies-literature