Recovery of tetrodotoxin from pufferfish viscera extract by amine-functionalized magnetic nanocomposites.

Tetrodotoxin (TTX) has been widely used in pharmacology, food poisoning analysis, therapeutic use, and neurobiology. In the last decades, the isolation and purification of TTX from natural sources (e.g., pufferfish) were mostly based on column chromatography. Recently, functional magnetic nanomaterials have been recognized as promising solid phases for the isolation and purification of bioactive compounds from aqueous matrices due to their effective adsorptive properties. Thus far, no studies have been reported on the utilization of magnetic nanomaterials for the purification of TTX from biological matrices. In this work, an effort has been made to synthesize Fe3O4@SiO2 and Fe3O4@SiO2-NH2 nanocomposites for the adsorption and recovery of TTX derivatives from a crude pufferfish viscera extract. The experimental data showed that Fe3O4@SiO2-NH2 displayed a higher affinity toward TTX derivatives than Fe3O4@SiO2, achieving maximal adsorption yields for 4epi-TTX, TTX, and Anh-TTX of 97.9, 99.6, and 93.8%, respectively, under the optimal conditions of contact time of 50 min, pH of 2, adsorbent dosage of 4 g L-1, initial adsorbate concentration of 1.92 mg L-1 4epi-TTX, 3.36 mg L-1 TTX and 1.44 mg L-1 Anh-TTX and temperature of 40 °C. Interestingly, desorption of 4epi-TTX, TTX, and Anh-TTX from Fe3O4@SiO2-NH2-TTX investigated at 50 °C was recorded to achieve the highest recovery yields of 96.5, 98.2, and 92.7% using 1% AA/ACN for 30 min reaction, respectively. Remarkably, Fe3O4@SiO2-NH2 can be regenerated up to three cycles with adsorptive performance remaining at nearly 90%, demonstrating a promising adsorbent for purifying TTX derivatives from pufferfish viscera extract and a potential replacement for resins used in column chromatography-based techniques.

Auxin production of the filamentous cyanobacterial Planktothricoides strain isolated from a polluted river in Vietnam.

Cyanobacteria are photosynthetic microorganisms with widespread diversity and extensive global distribution. They produce a wide variety of bioactive substances (e.g., lipopeptides, fatty acids, toxins, carotenoids, vitamins and plant growth regulators) that are released into culture media. In this study, the capability of a cyanobacterial strain of Planktothricoides raciborskii to produce intra- and extracellular auxins was investigated. The filamentous cyanobacterial P. raciborskii strain was isolated from a river in Vietnam, and it was cultivated in the laboratory under the optimum conditions of the BG11 culture medium and a pH of 7.0. The auxins were identified and quantified by the Salkowski colorimetric method and high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS). Colorimetric analysis revealed that P. raciborskii produces extracellular indole-3-acetic acid (IAA) in the absence and presence of l-tryptophan. The maximum extracellular IAA concentration of the culture reached 118 ± 2 µg mL-1, which was supplemented with 900 µg mL-1 of l-tryptophan. HPLC-MS analysis revealed that the isolated cyanobacteria accumulate other plant-growth-promoting hormones besides IAA, such as indole-3-carboxylic acid (ICA), indole-3 butyric acid (IBA) and indole propionic acid (IPA). This is the first report on the production of auxins in an isolated strain of cyanobacteria Planktothricoides from a polluted river. The capability of producing auxins makes the P. raciborskii strain an appropriate candidate for the formulation of a biofertilizer.

Kinetics of enzymatic transesterification and thermal deactivation using immobilized Burkholderia lipase as catalyst.

The most effective way of enzymatic synthesis of biodiesel is through lipase-catalyzed transesterification, while its performance and economic feasibility should still be improved. In this study, lipase produced by an isolated Burkholderia sp. was immobilized on microsize Celite materials functionally modified with long alkyl groups. The specific activity of the immobilized lipase was 1,154 U/g. The methanolysis of olive oil catalyzed by the immobilized lipase obeyed Ping Pong Bi Bi model with an estimated V max, K m,TG, K m,M and K i,M value of 0.61 mol/(L min), 7.93 mol/L, 1.01 mol/L, and 0.24 mol/L, respectively. The activation energy of the enzymatic reaction is estimated as 15.51 kJ/mol. The immobilized lipase exhibits high thermal stability with thermal deactivation energy of 83 kJ/mol and a long half-life. The enthalpy, Gibb's free energy, and entropy of the immobilized lipase were in the range of 80.02-80.35 kJ/mol, 88.35-90.13 kJ/mol, and -28.22 to -25.11 J/(mol K), respectively.

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.

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.

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.

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.

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.

Response surface optimization of dissolved oxygen and nitrogen sources for the biodegradation of MTBE and BTEX.

Response surface methodology (RSM) using central composite design was applied to obtain the optimal dissolved oxygen (DO) and nitrogen (N) concentrations for biodegrading MTBE (Methyl tert-butyl ether) and BTEX (benzene, ethylbenzene, toluene, p-xylene). Moreover, the effects of DO, N, and their interaction on the degradation process were evaluated. It was found that N, N(2), DO and DO(2) have significant effects on the efficiency of MTBE and BTEX removal. The removal efficiency when using biostimulation with bioaugmentation (BwB) is higher than with other processes, being greater than 82% at concentrations of 12 and 48 mg l(-1) for DO and N, respectively. However, it was also found that the interaction term of DO x N has no significant effect on the degradation processes.