Search results for: methanolysis
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 3

Search results for: methanolysis

3 In situ Real-Time Multivariate Analysis of Methanolysis Monitoring of Sunflower Oil Using FTIR

Authors: Pascal Mwenge, Tumisang Seodigeng

Abstract:

The combination of world population and the third industrial revolution led to high demand for fuels. On the other hand, the decrease of global fossil 8fuels deposits and the environmental air pollution caused by these fuels has compounded the challenges the world faces due to its need for energy. Therefore, new forms of environmentally friendly and renewable fuels such as biodiesel are needed. The primary analytical techniques for methanolysis yield monitoring have been chromatography and spectroscopy, these methods have been proven reliable but are more demanding, costly and do not provide real-time monitoring. In this work, the in situ monitoring of biodiesel from sunflower oil using FTIR (Fourier Transform Infrared) has been studied; the study was performed using EasyMax Mettler Toledo reactor equipped with a DiComp (Diamond) probe. The quantitative monitoring of methanolysis was performed by building a quantitative model with multivariate calibration using iC Quant module from iC IR 7.0 software. 15 samples of known concentrations were used for the modelling which were taken in duplicate for model calibration and cross-validation, data were pre-processed using mean centering and variance scale, spectrum math square root and solvent subtraction. These pre-processing methods improved the performance indexes from 7.98 to 0.0096, 11.2 to 3.41, 6.32 to 2.72, 0.9416 to 0.9999, RMSEC, RMSECV, RMSEP and R2Cum, respectively. The R2 value of 1 (training), 0.9918 (test), 0.9946 (cross-validation) indicated the fitness of the model built. The model was tested against univariate model; small discrepancies were observed at low concentration due to unmodelled intermediates but were quite close at concentrations above 18%. The software eliminated the complexity of the Partial Least Square (PLS) chemometrics. It was concluded that the model obtained could be used to monitor methanol of sunflower oil at industrial and lab scale.

Keywords: biodiesel, calibration, chemometrics, methanolysis, multivariate analysis, transesterification, FTIR

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2 Lipid from Activated Sludge as a Feedstock for the Production of Biodiesel

Authors: Ifeanyichukwu Edeh, Tim Overton, Steve Bowra

Abstract:

There is increasing interest in utilising low grade or waste biomass for the production of renewable bioenergy vectors i.e. waste to energy. In this study we have chosen to assess, activated sludge, which is a microbial biomass generated during the second stage of waste water treatment as a source of lipid for biodiesel production. To date a significant proportion of biodiesel is produced from used cooking oil and animal fats. It was reasoned that if activated sludge proved a viable feedstock it has the potential to support increase biodiesel production capacity. Activated sludge was obtained at different times of the year and from two different sewage treatment works in the UK. The biomass within the activated sludge slurry was recovered by filtration and the total weight of material calculated by combining the dry weight of the total suspended solid (TSS) and the total dissolved solid (TDS) fractions. Total lipids were extracted from the TSS and TDS using solvent extraction (Folch methods). The classes of lipids within the total lipid extract were characterised using high performance thin layer chromatography (HPTLC) by referencing known standards. The fatty acid profile and content of the lipid extract were determined using acid mediated-methanolysis to obtain fatty acid methyl esters (FAMEs) which were analysed by gas chromatography and HPTLC. The results showed that there were differences in the total biomass content in the activated sludge collected from different sewage works. Lipid yields from TSS obtained from both sewage treatment works differed according to the time of year (between 3.0 and 7.4 wt. %). The lipid yield varied slightly within the same source of biomass but more widely between the two sewage treatment works. The neutral lipid classes identified were acylglycerols, free fatty acids, sterols and wax esters while the phospholipid class included phosphatidylcholine, lysophosphatidycholine, phosphatidylethanolamine and phosphatidylinositol. The fatty acid profile revealed the presence of palmitic acid, palmitoleic acid, linoleic acid, oleic acid and stearic acid and that unsaturated fatty acids were the most abundant. Following optimisation, the FAME yield was greater than 10 wt. % which was required to have an economic advantage in biodiesel production.

Keywords: activated sludge, biodiesel, lipid, methanolysis

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1 Surface Display of Lipase on Yarrowia lipolytica Cells

Authors: Evgeniya Y. Yuzbasheva, Tigran V. Yuzbashev, Natalia I. Perkovskaya, Elizaveta B. Mostova

Abstract:

Cell-surface display of lipase is of great interest as it has many applications in the field of biotechnology owing to its unique advantages: simplified product purification, and cost-effective downstream processing. One promising area of application for whole-cell biocatalysts with surface displayed lipase is biodiesel synthesis. Biodiesel is biodegradable, renewable, and nontoxic alternative fuel for diesel engines. Although the alkaline catalysis method has been widely used for biodiesel production, it has a number of limitations, such as rigorous feedstock specifications, complicated downstream processes, including removal of inorganic salts from the product, recovery of the salt-containing by-product glycerol, and treatment of alkaline wastewater. Enzymatic synthesis of biodiesel can overcome these drawbacks. In this study, Lip2p lipase was displayed on Yarrowia lipolytica cells via C- and N-terminal fusion variant. The active site of lipase is located near the C-terminus, therefore to prevent the activity loosing the insertion of glycine-serine linker between Lip2p and C-domains was performed. The hydrolytic activity of the displayed lipase reached 12,000–18,000 U/g of dry weight. However, leakage of enzyme from the cell wall was observed. In case of C-terminal fusion variant, the leakage was occurred due to the proteolytic cleavage within the linker peptide. In case of N-terminal fusion variant, the leaking enzyme was presented as three proteins, one of which corresponded to the whole hybrid protein. The calculated number of recombinant enzyme displayed on the cell surface is approximately 6–9 × 105 molecules per cell, which is close to the theoretical maximum (2 × 106 molecules/cell). Thus, we attribute the enzyme leakage to the limited space available on the cell surface. Nevertheless, cell-bound lipase exhibited greater stability to short-term and long-term temperature treatment than the native enzyme. It retained 74% of original activity at 60°C for 5 min of incubation, and 83% of original activity after incubation at 50°C during 5 h. Cell-bound lipase had also higher stability in organic solvents and detergents. The developed whole-cell biocatalyst was used for recycling biodiesel synthesis. Two repeated cycles of methanolysis yielded 84.1–% and 71.0–% methyl esters after 33–h and 45–h reactions, respectively.

Keywords: biodiesel, cell-surface display, lipase, whole-cell biocatalyst

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