Semi-rational Engineering of a Promiscuous Fatty Acid Hydratase for Alteration of Regioselectivity

Fatty acid hydratases (FAHs) catalyze regio- and stereo-selective hydration of unsaturated fatty acids to produce hydroxy fatty acids. Fatty acid hydratase-1 (FA-HY1) from Lactobacillus Acidophilus is the most promiscuous and regiodiverse FAH identified so far. Here, we engineered binding site residues of FA-HY1 (S393, S395, S218 and P380) by semi-rational protein engineering to alter regioselectivity. Although it was not possible to obtain a completely new type of regioselectivity with our mutant libraries, a significant shift of regioselectivity was observed towards cis-5, cis-8, cis-11, cis-14, cis-17-eicosapentaenoic acid (EPA). We identified mutants (S393/S395 mutants) with excellent regioselectivity, generating a single hydroxy fatty acid product from EPA (15-OH product), which is advantageous from application perspective. This result is impressive given that wild-type FA-HY1 produces a mixture of 12-OH and 15-OH products at 63 : 37 ratio (12-OH : 15-OH). Moreover, our results indicate that native FA-HY1 is at its limit in terms of promiscuity and regiospecificity, thus it may not be possible to diversify its product portfolio with active site engineering. This behavior of FA-HY1 is unlike its orthologue, fatty acid hydratase-2 (FA-HY2; 58 % sequence identity to FA-HY1), which has been shown earlier to exhibit significant promiscuity and regioselectivity changes by a few active site mutations. Our reverse engineering from FA-HY1 to FA-HY2 further demonstrates this conclusion.     

The oxidation of NaBH4 on electrochemicaly treated silver electrodes

This study is related to the development of suitable anode material for the direct borohyride fuel cells (DBFCs). The effect of electroactive Ag oxides upon the oxidation of NaBH₄ was investigated. The treatment of Ag surface with H₂O₂ gave a granulated compact layer which was observed to give a very high current density and good reversibility at lower concentration compared with the electrode treated with NaOH which resulted a fibrous layer with a lower electrocatalytic effect. The mode of surface oxidation was found to have a profound effect upon the behavior of the electrode. It was also observed that there is an optimum NaBH₄ concentration to obtain the maximum current density.     

Counterion-coupled gemini (Cocogem) surfactants based on dodecylisopropylol amine and dicarboxylic acids: synthesis,characterization and evaluation as biocide against SRB

New cocogem surfactants were synthesized by interaction of dodecylisopropylol amine with dicarboxylic (oxalic, succinic, adipic, sebacic, tartaric, maleic, fumaric, isophthalic) acids. By tensiometric method, the surface activity of aqueous solutions of the synthesized cocogem surfactants at the border with air was studied and, by conductometric method, the specific electrical conductivity of these solutions was determined. The degree of counterion binding (β), critical micelle concentration (CMC), effectiveness of surface tension reduction (π_CMC), surface excess concentration (Γ_max), area per molecule at the interface (A_min), changes of Gibbs free energies of adsorption (ΔG_ad) and micellization (ΔG_mic) have been calculated. The character of change of the colloidal-chemical indices depending on spacer-group nature and length has been clarified. So, with an elongation of the spacer group and when passing from cis-form to trans-form, the value of CMC decreases. The obtained cocogem surfactants exhibit a considerable bactericidal effectiveness against sulfate-reducing bacteria. The bactericide properties of the cocogem surfactants containing in the spacer chain a saturated hydrocarbon fragment and benzene ring are stronger than for the others.     

Synthesis and characterization of novel polyfuran/poly(2‐iodoaniline) conducting composite

The electrically conducting polyfuran/poly(2‐iodoaniline) (PFu/P2IAn) and P2IAn/PFu composites were prepared by chemical oxidative polymerization using polyfuran and poly(2‐iodoaniline) in HCl and CHCl₃ media. The conductivities of composites were determined as a function of the amount of guest polymer. It was found out that the conductivities of P2IAn/PFu composites increased 100‐fold, whereas the conductivities of PFu/P2IAn composites did not show a specific increase. The composite compositions were altered by varying guest polymer feed ratios during preparation. Generally, the electrical conductivities of P2IAn/PFu composites increased with increasing the amount of PFu. Homopolymers and composites were further characterized thermally, employing thermogravimetry (TGA) and morphologically employing scanning electron microscopy (SEM). Further evidences concerning the polymer structures were obtained by FTIR and UV‐vis spectroscopies and magnetic susceptibility measurements. TGA results revealed that PFu/P2IAn among the homopolymers (PFu and P2IAn) and P2IAn/PFu composite have the highest thermal stability. The composites synthesized varying the host and the guest polymer order have different conductivities, morphological structures, and thermal properties. © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 89: 2823–2830, 2003     

Metal chelating properties of Cibacron Blue F3GA‐derived poly(EGDMA‐HEMA) microbeads

Metal chelating properties of Cibacron Blue F3GA‐derived poly(EGDMA‐HEMA) microbeads have been studied. Poly(EGDMA‐HEMA) microbeads were prepared by suspension copolymerization of ethylene glycol dimethacrylate (EGDMA) and hydroxy‐ethyl methacrylate (HEMA) by using poly(vinyl alcohol), benzoyl peroxide, and toluene as the stabilizer, the initiator, and the pore‐former, respectively. Cibacron Blue F3GA was covalently attached to the microbeads via the nucleophilic substitution reaction between the chloride of its triazine ring and the hydroxyl groups of the HEMA, under alkaline conditions. Microbeads (150–200 μm in diameter) with a swelling ratio of 55%, and carrying 16.5 μmol Cibacron Blue F3GA/g polymer were used in the adsorption/desorption studies. Adsorption capacity of the microbeads for the selected metal ions, i.e., Cu(II), Zn(II), Cd(II), Fe(III), and Pb(II) were investigated in aqueous media containing different amounts of these ions (5–200 ppm) and at different pH values (2.0–7.0). The maximum adsorptions of metal ions onto the Cibacron Blue F3GA‐derived microbeads were 0.19 mmol/g for Cu(II), 0.34 mmol/g for Zn(II), 0.40 mmol/g for Cd(II), 0.91 mmol/g for Fe(III), and 1.05 mmol/g for Pb(II). Desorption of metal ions were studied by using 0.1 M HNO₃. High desorption ratios (up to 97%) were observed in all cases. Repeated adsorption/desorption operations showed the feasibility of repeated use of this novel sorbent system. © 1999 John Wiley & Sons, Inc. J Appl Polym Sci 71: 1397–1403, 1999     

Comparison of metal chelate affinity sorption of BSA onto Dye/Zn (II)-derived poly(ethylene glycol dimethacrylate-hydroxyethyl methacrylate) microbeads

Poly(ethylene glycol dimethacrylate-hydroxyethyl methacrylate) [poly-(EGDMA-HEMA)]microbeads in the size range of 150–200 μm were produced by a modified suspension copolymerization of EGDMA and HEMA. The dyes (Congo red, Cibacron blue F3GA, and alkali blue 6B) were covalently immobilized; then, Zn(II) ions were incorporated within the microbeads by chelation with the dye molecules. The maximum amounts of dye loadings were 14.5 μmol/g, 16.5 μmol/g, and 23.7 μmol/g for Congo red, Cibacron blue F3GA, and alkali blue 6B, respectively. Different amounts of Zn(II) ions(2.9–53.8 mg/g polymer) were incorporated on the microbeads by changing the initial concentration of Zn(II) ions and the pH of the medium. Bovine serum albumin (BSA) adsorption onto dye/Zn(II)-derived microbeads containing Congo red, Cibacron blue F3GA, and alkali blue 6B was investigated. The maximum BSA adsorptions onto the dye/Zn(II)-derived microbeads from aqueous solutions containing different amounts of BSA were 159 mg/g, 122 mg/g, and 93 mg/g for the Congo red, Cibacron blue F3GA, and alkali blue 6B dyes, respectively. The maximum BSA adsorptions were observed at pH 6.0 in all cases. Desorption of BSA molecules was achieved by using 0.025M EDTA (pH 4.9). High desorption ratios (more than 93% of the adsorbed BSA) were observed in all cases. It was possible to reuse these novel metal chelate sorbents without significant losses in their adsorption capacities. © 1997 John Wiley & Sons, Inc. J Appl Polym Sci 65: 2085–2093, 1997     

A review on the performance of photovoltaic/thermoelectric hybrid generators

Utilization of a broad range of solar spectrum has the potential for high power output from solar cells. However, solar photovoltaics (PVs) can convert only part of the solar electromagnetic spectrum into electricity efficiently. The remaining of the solar radiation is often dissipated in the form of heat, which causes performance reduction and reduces the life expectancy of the solar PV cell. Thermoelectric generators (TEGs) are devices that operate like a heat engine by converting thermal energy into electricity through thermoelectric effect. Integrating a TEG into a PV converter will enhance its efficiency and reduce the amount of heat dissipated. Different studies have been carried out and are still taking place to increase the total efficiency of a coupled photovoltaic thermoelectric generator (PV-TEG) system. This review discusses the concept of PV converters and thermoelectric devices and presents the various models and numerical and experimental investigations on performance enhancement of integrated PV-TEGs. The influence of key parameters on the performance of PV-TEG were also discussed. The review is expected to serve as a reference to recent work on research and development of integrated PV-TEG systems.     

Mass transfer enhancement factor for chemical absorption of carbon dioxide into sodium metaborate solution

Hydrogen is getting increasing attention as a medium for energy storage, and sodium borohydride is accepted as a suitable carrier for hydrogen. The main product of the process by means of which hydrogen is produced from sodium borohydride is sodium metaborate. Our aim was to find an alternative use for sodium metaborate and specifically investigating the feasibility to use it for carbon dioxide capture from flue gases. The products of this chemical absorption are sodium carbonate, sodium bicarbonate and boric acid, all of which are industrially important chemicals. A bubble column was used in the experiments. Oxygen desorption technique was employed to determine the liquid side physical mass transfer coefficient. Chemical mass transfer coefficient was determined by absorption of carbon dioxide from its mixture with nitrogen into sodium metaborate solution. Enhancement factor was then calculated and a correlation was developed for it.     

Rational Engineering of Hydratase from Lactobacillus acidophilus Reveals Critical Residues Directing Substrate Specificity and Regioselectivity

Enzymatic conversion of fatty acids (FAs) by fatty acid hydratases (FAHs) presents a green and efficient route for high-value hydroxy fatty acid (HFA) production. However, limited diversity was achieved among HFAs, to date, with respect to chain length and hydroxy position. In this study, two highly similar FAHs from Lactobacillus acidophilus were compared: FA-HY2 has a narrow substrate scope and strict regioselectivity, whereas FA-HY1 utilizes longer chain substrates and hydrates various double-bond positions. It is revealed that three active-site residues play a remarkable role in directing substrate specificity and regioselectivity of hydration. If these residues on FA-HY2 are mutated to the corresponding ones in FA-HY1, a significant expansion of substrate scope and a distinct enhancement in hydration of double bonds towards the ω-end of FAs is observed. A three-residue mutant of FA-HY2 (TM-FA-HY2) displayed an impressive reversal of regioselectivity towards linoleic acid, shifting the ratio of the HFA regioisomers (10-OH/13-OH) from 99:1 to 12:88. Notable changes in regioselectivity were also observed for arachidonic acid and for C₁₈ polyunsaturated fatty acid substrates. In addition, TM-FA-HY2 converted eicosapentaenoic acid into its 12-hydroxy product with high conversion at the preparative scale. Furthermore, it is demonstrated that microalgae are a source of diverse FAs for HFA production. This study paves the way for tailor-made FAH design to enable the production of diverse HFAs for various applications from the polymer industry to medical fields.     

Experimental study for predicting the specific heat of water based Cu-Al2O3 hybrid nanofluid using artificial neural network and proposing new correlation

In this study, an artificial neural network model has been created in order to estimate the specific heat of Cu-Al₂O₃/water hybrid nanofluid based on temperature (T) and volume concentration (φ). Specific heat values of the Cu-Al₂O₃/water hybrid nanofluid prepared in five-volume concentration were measured experimentally in the 20°C to 65°C temperature range. The dataset was reserved into three primary parts, with the inclusion of 901 (70%) for the training, 257 (20%) for the test and 129 (10%) for the validation. As a result of comparison with experimental values, it is concluded that this model predicts specific heat with R-value of 0.99994 and an average relative error of approximately 5.84e-9. In addition, a mathematical correlation has been developed to estimate the specific heat of the Cu-Al₂O₃/water hybrid nanofluid. The data acquired from the mathematical correlation, developed, were in great correlation with all the experimental values with an average deviation of −0.005%. This result has revealed that the developed mathematical correlation is an ideal design for estimating the specific heat of the Cu-Al₂O₃/water hybrid nanofluid.