Dang-Thuan Tran

Enzymatic transesterification of microalgal oil from Chlorella vulgaris ESP-31 for biodiesel synthesis using immobilized Burkholderia lipase.

An indigenous microalga Chlorella vulgaris ESP-31 grown in an outdoor tubular photobioreactor with CO(2) aeration obtained a high oil content of up to 63.2%. The microalgal oil was then converted to biodiesel by enzymatic transesterification using an immobilized lipase originating from Burkholderia sp. C20. The conversion of the microalgae oil to biodiesel was conducted by transesterification of the extracted microalgal oil (M-I) and by transesterification directly using disrupted microalgal biomass (M-II). The results show that M-II achieved higher biodiesel conversion (97.3 wt% oil) than M-I (72.1 wt% oil). The immobilized lipase worked well when using wet microalgal biomass (up to 71% water content) as the oil substrate. The immobilized lipase also tolerated a high methanol to oil molar ratio (>67.93) when using the M-II approach, and can be repeatedly used for six cycles (or 288 h) without significant loss of its original activity.

Immobilization of Burkholderia sp. lipase on a ferric silica nanocomposite for biodiesel production.

In this work, lipase produced from an isolated strain Burkholderia sp. C20 was immobilized on magnetic nanoparticles to catalyze biodiesel synthesis. Core-shell nanoparticles were synthesized by coating Fe(3)O(4) core with silica shell. The nanoparticles treated with dimethyl octadecyl [3-(trimethoxysilyl) propyl] ammonium chloride were used as immobilization supporters. The Burkholderia lipase was then bound to the synthesized nanoparticles for immobilization. The protein binding efficiency on alkyl-functionalized Fe(3)O(4)-SiO(2) was estimated as 97%, while the efficiency was only 76% on non-modified Fe(3)O(4)-SiO(2). Maximum adsorption capacity of lipase on alkyl-functionalized Fe(3)O(4)-SiO(2) was estimated as 29.45 mg g(-1) based on Langmuir isotherm. The hydrolytic kinetics (using olive oil as substrate) of the lipase immobilized on alkyl-grafted Fe(3)O(4)-SiO(2) followed Michaelis-Menten model with a maximum reaction rate and a Michaelis constant of 6251 Ug(-1) and 3.65 mM, respectively. Physical and chemical properties of the nanoparticles and the immobilized lipase were characterized by Brunauer-Emmett-Teller (BET) analysis, scanning electron microscope (SEM), and Fourier transform infrared spectroscopy (FT-IR). Moreover, the immobilized lipase was used to catalyze the transesterification of olive oil with methanol to produce fatty acid methyl esters (FAMEs), attaining a FAMEs conversion of over 90% within 30 h in batch operation when 11 wt% immobilized lipase was employed. The immobilized lipase could be used for ten cycles without significant loss in its transesterification activity.

Effect of solvents and oil content on direct transesterification of wet oil-bearing microalgal biomass of Chlorella vulgaris ESP-31 for biodiesel synthesis using immobilized lipase as the biocatalyst.

In this work, a one-step extraction/transesterification process was developed to directly convert wet oil-bearing microalgal biomass of Chlorella vulgaris ESP-31 into biodiesel using immobilized Burkholderia lipase as the catalyst. The microalgal biomass (water content of 86-91%; oil content 14-63%) was pre-treated by sonication to disrupt the cell walls and then directly mixed with methanol and solvent to carry out the enzymatic transesterification. Addition of a sufficient amount of solvent (hexane is most preferable) is required for the direct transesterification of wet microalgal biomass, as a hexane-to-methanol mass ratio of 1.65 was found optimal for the biodiesel conversion. The amount of methanol and hexane required for the direct transesterification process was also found to correlate with the lipid content of the microalga. The biodiesel synthesis process was more efficient and economic when the lipid content of the microalgal biomass was higher. Therefore, using high-lipid-content microalgae as feedstock appears to be desirable.

Biodiesel synthesis via heterogeneous catalysis using modified strontium oxides as the catalysts

In this work, alkaline earth metal oxides (i.e., MgO, CaO, and SrO) were used as catalysts for the transesterification of olive oil with methanol. The most efficient catalyst was further doped with either CaO or SiO(2) to improve its catalytic activity, which was evaluated by conducting transesterification at different reaction temperatures, different water content, and using different types of oils. Finally, repeated tests were conducted to evaluate the reusability of the doped catalyst. The results show that the conversion of refined olive oil to biodiesel was more than 80% in 15 min when SrO was applied, while using SrO doped SiO(2) (SrO/SiO(2)) further increased the conversion to 95% in 10 min. SrO/SiO(2) also featured good water and free fatty acids (FFAs) tolerance, as the conversion was still higher than 90% (in 20 min) when the water and FFAs contents were increased to 3.23 and 3.14 wt.%, respectively. Addition of hexane significantly improved the reusability of SrO/SiO(2) for transesterification, as the biodiesel production still reached nearly 80% after the catalyst was repeatedly used for four times.

Microalgae harvesting and subsequent biodiesel conversion.

Chlorella vulgaris ESP-31 containing 22.7% lipid was harvested by coagulation (using chitosan and polyaluminium chloride (PACl) as the coagulants) and centrifugation. The harvested ESP-31 was directly employed as the oil source for biodiesel production via transesterification catalyzed by immobilized Burkholderia lipase and by a synthesized solid catalyst (SrO/SiO2). Both enzymatic and chemical transesterification were significantly inhibited in the presence of PACl, while the immobilized lipase worked well with wet chitosan-coagulated ESP-31, giving a high biodiesel conversion of 97.6% w/w oil, which is at a level comparable to that of biodiesel conversion from centrifugation-harvested microalgae (97.1% w/w oil). The immobilized lipase can be repeatedly used for three cycles without significant loss of its activity. The solid catalyst SrO/SiO2 worked well with water-removed centrifuged ESP-31 with a biodiesel conversion of 80% w/w oil, but the conversion became lower (55.7-61.4% w/w oil) when using water-removed chitosan-coagulated ESP-31 as the oil source.

Kinetics of transesterification of olive oil with methanol catalyzed by immobilized lipase derived from an isolated Burkholderia sp. strain.

This work was carried out to investigate the acyl migration phenomena which has been considered as the factor having significant impact on kinetics of transesterification of oils catalyzed by a Burkholderia lipase with 1,3-regioselectivity. Transesterification of olive oil with methanol catalyzed by the immobilized lipase produces various intermediates, including 1-monoglyceride, 2-monoglyceride, 1,2-diglyceride, and 1,3-diglyceride. Migration kinetics of fatty acid groups from sn-2 of 2-monoglyceride and 1,2-diglyceride to 1-monoglyceride and 1,3-diglyceride were investigated for the temperature range of 25-65°C. The kinetics of transesterification of olive oil with methanol involving acyl migration in the presence of water was also systematically studied at 25, 40, and 65°C. Increasing temperature could increase the acyl migration rate. The overall biodiesel conversion was improved from 73.4% (at 25°C) to 90.0% and 92.4% when conducting at 40 and 65°C, respectively. Thermodynamics aspects of equilibrium state of the immobilized lipase-catalyzed transesterification were also discussed.