Evaluation and optimization of food-grade tannin acyl hydrolase production by a probiotic Lactobacillus plantarum strain in submerged and solid state fermentation

Tannin acyl hydrolase (tannase) production by Lactobacillus plantarum MTCC1407 was studied in submerged and solid-state fermentation process. Sequential optimization strategy using Plackett–Burman screening and response surface methodology was adopted to optimize the submerged fermentation process. Eight medium components were evaluated initially by Plackett–Burman two level factorial designs to identify the most significant parameters that affect the tannase production. The significant variables affecting tannase production were found to be tannic acid, glucose and MnSO₄·7H₂O. These factors were further optimized by response surface methodology. Maximum tannase activity of 9.13 U ml^?1 was observed at 30 h using the following medium composition (g l^?1): tannic acid, 13.16; glucose, 1.5; NH₄Cl, 1.0; CaCl₂·2H₂O, 1.0; K₂HPO₄, 0.5; KH₂PO₄, 0.5; MgSO4·7H₂O, 0.5 and MnSO₄·7H₂O, 0.03. Among the various carbon sources examined for tannase production by L. plantarum, glucose and tannic acid combination was found to be decisive for enhancing tannase yield. Solid state fermentation was conducted using various solid substrates and agricultural residues. Maximum tannase activity of 5.319 U gds^?1 was obtained using coffee husk as substrate.     

Optimization of critical medium components using response surface methodology for lipase production by Rhizopus delemar

The production process of a 1,3-position specific lipase from Rhizopus delemar was optimized by response surface methodology (RSM) and a Box–Behnken experimental design was used to study the interactive effects of fermentation medium components on lipase activity and microorganism growth. Preliminary batch tests were employed to obtain the favorable conditions for lipase activity analysis and found that sucrose, molasses, yeast extract, sunflower oil, tween-80 have significant influences on the lipase production and microorganism growth. The concentrations of five fermentation medium components were optimized. Among five variables, molasses sucrose and yeast extract were identified as less significant variables for lipase production. The optimum fermentation medium composition for lipase production by R. delemar was sucrose concentration 4.19 g/L, molasses sucrose 1.32 g/L, yeast extract 0.53 g/L, sunflower oil 1.11% (v/v), and tween-80 1.80% (v/v). In these conditions, the biomass concentration of 4.52 g/L with a lipolytic activity of 1585 μmol/L min was reached.     

Influence of the bed geometry on the kinetics of the extraction of clove bud oil with supercritical CO2

There is a need for scientific research that evaluates the influence of important process variables on the scale up of supercritical technology. For supercritical fluid extraction (SFE), one of these variables is the extractor's bed geometry, which can be defined by the ratio of the bed height (H_B) to the bed diameter (D_B). A systematic study is needed to select suitable criteria that can be used to obtain similar extraction curves among beds with different geometries. In this study, maintaining a constant ratio of solvent mass to feed mass for two beds with 1-L volumes but different geometries (E-1: H_B/D_B = 7.1; E-2: H_B/D_B = 2.7) was confirmed as a successful scale up criterion. For constant values of the temperature, pressure and bed porosity, there is experimental evidence that the mass transfer rate is equal in the two beds when the solvent flow rate is high. When 0.6 kg of clove buds was packed in the beds, the extraction rates were 2.10 ± 0.08 and 2.3 ± 0.1 g extract/min for beds E-1 and E-2, respectively. However, when the solvent flow rate was lower, the extraction rates were 0.93 ± 0.06 and 1.12 ± 0.02 g extract/min for beds E-1 and E-2, respectively. This difference in behavior between the extraction beds is associated with the axial dispersion of the fluid, which is more pronounced when the H_B/D_B ratio is increased. Thin particles tend to compact in the beds with high H_B/D_B ratios, which shorten the solvent passage. Non-isothermal profiles and differences in chemical composition of the extracts were also observed: 17% more α-humulene and 9% more eugenol were extracted in E-1 and E-2, respectively.     

Characterization of purified fungal endoxylanase and its application for production of value added food ingredient from agroresidues

Xylanase from Aspergillus foetidus MTCC 4898 was purified using ammonium sulphate fractionation followed by ion exchange and gel permeation chromatography with 12.26-fold purity and 29.9% recovery. Purified xylanase was found to be 29.9 kDa. Optimum temperature and pH for xylanase activity of purified xylanase were found to be 50 °C and 5.3 respectively. Presence of additives like polyethylene glycol, sodium azide, Tween 80, KCl and NaCl increased the stability of purified xylanase by 35, 29, 28, 32 and 43% respectively at 50 °C after 180 min. Kinetic parameters like K_m and V_max were found to be 4 mg/ml and 7288 μmol/mg/min respectively. The purified xylanase was found to be an endoxylanase as it produced only xylooligosaccharides (XOS) from birchwood xylan. Production of XOS was carried out from xylan extracted from agro-residues using β-xylosidase free xylanase. Maximum yield of XOS was 7.28 ± 0.14 mg/ml and 4.52 ± 0.21 mg/ml from wheat straw xylan and rice straw xylan respectively. XOS mixture was suitable for food industry looking at its high thermal stability at low pH. Prebiotic effect of XOS was evaluated by in vitro fermentation of XOS using known probiotic strains of Bifidobacterium spp.     

Solution and structural binding studies of phosphate with thiophene-based azamacrocycles

Two thiophene-based monocyclic receptors L1 and L2 have been studied for phosphate binding in solutions (D₂O and DMSO-d₆) by ¹H NMR and ³¹P NMR titrations, and in the solid state by single crystal X-ray analysis. Results from ¹H NMR titrations suggest that the ligands bind phosphate anions in a 1:2 binding mode in DMSO-d₆, with the binding constants of 5.25 and 4.20 (in log K), respectively. The binding of phosphate to L1 and L2 was further supported by ³¹P NMR in D₂O at pH = 5.2. The crystal structure of the phosphate complex of L1 reveals unambiguous proof for the formation of a ditopic complex via multiple hydrogen bonds from NH ⋯ O and CH ⋯ O interactions.     

Fermentation and heat-moisture treatment induced changes on the physicochemical properties of foxtail millet (Setaria italica) flour

This research intends to verify the effect of Lactobacillus paracasei Fn032 fermentation and heat-moisture treatment (HMT) on the physicochemical properties of foxtail millet (Setaria italica) flour. The results obtained showed a significant (P < 0.05) increase in protein content (12.02–20.54%), total starch (15.78–51.01%) and starch fractions after fermentation and HMT. Differential scanning calorimetry (DSC) analysis showed high decomposition temperature (T_d) trend of 180.59 and 189.82 °C after HMT. However, there was significant (P < 0.05) enthalpy (ΔH) decrease. Flour digestion resulted in variation of slow digestible starch (SDS) and resistant starch (RS) count from 6.83 to 18.42% and 7.61 to 22.68% respectively, after fermentation and HTM. Following this observation, it was ascertained that in X-ray diffraction; pasting viscosity and fluorescence spectrophotometry show greater HMT influenced on the flour components. Findings from the scanning electron microscopy (SEM) analysis showed microstructure differences of the flours samples. Fermentation and heat moisture treatment methods present a possible way of changing or improving the physicochemical properties and add nutritional value to foxtail millet meal.     

Ammonia removal by air stripping in a semi-batch jet loop reactor

Ammonia is very toxic chemical and it can be removed by air stripping at high pH. JLRs have found applications in wastewater treatment processes due to their high mass transfer rates. In JLRs, intrinsic high turbulence result in a very large air-liquid surface area for greater mass transfer. Therefore, in this study, ammonia removal by air stripping from synthetically prepared ammonia solution at the high pH in a semi-batch JLR due to its high mass transfer capabilities have been investigated. Investigated parameters in a JLR were initial ammonia concentration (10–500 mg/L), temperature (20–50 °C), air flow rate (5–50 L/min) and liquid circulation rate (35–50 L/min). While it was demonstrated that temperature and air flow rate have a significant effect on the ammonia removal, it was determined that initial ammonia concentration and liquid circulation rate have no significant effect on the ammonia removal. The overall volumetric mass transfer coefficients (K_La) have been calculated from obtained model and it was determined that increasing temperature and air flow rate have a very significant effect on K_La. It was concluded that JLR provides higher mass transfer capabilities than other type of reactors even if less air is given.     

Formation of diamond-like BN phases under shock compression of graphite-like BN with different degree of structural ordering

The formation of diamond-like modifications of BN under high-temperature shock compression (P = 33 GPa and T up to 3500 K) has been studied. The powders of graphite-like BN with different degree of three-dimensional ordering of structure (D₃) were used as starting materials. To increase shock temperature and preserve dense phase formed, boron nitride powder mixed with alkali halide salt was compressed in cylindrical ampoule. The recovered samples were studied by X-ray diffraction and electron microscopy. It was revealed that the phase composition of shock compression products depends on the degree of three-dimensional ordering of initial graphite-like structure. Turbostratic BN with disordered structure (D₃ = 0) transformed mainly into cubic cBN by diffusion mechanism, whereas highly ordered graphite-like BN (D₃ = 0.95) transformed only into wurtzite wBN by martensitic mechanism. Both cubic and wurtzite dense modifications are formed from partly ordered initial structures (for which D₃ was 0.45 ÷ 0.7). The both diamond-like phases have nanocrystalline structure.     

Influence of the pH on the ZnO nanoparticle growth in supercritical water: Experimental and simulation approaches

In order to improve the knowledge on the nucleation and the growth mechanisms of metal oxides nanoparticles produced in supercritical water domain, ZnO was used as a “model” material. A continuous process of hydrothermal synthesis was employed to synthesize ZnO nanopowders (T = 410 °C and P = 305 bar) from Zn(NO₃)₂ and KOH solutions with different values of [KOH]/[Zn(II)] ratio from 0 to 8 in order to investigate the pH effect on the growth of ZnO nanocrystallite in terms of size and morphology. The samples were characterized by X-Ray Diffraction and Transmission Electronic Microscopy. ZnO crystal was considered as a cylindrical crystallite with a diameter D and height H. Especially, the aspect ratio D/H was already used to observe the change of ZnO nanoparticle shape correlated with TEM observations and results from a CFD simulation model. A schematic synoptic of the ZnO growth in supercritical domain is also presented.     

Assembly of one 2D layer and two 3D pillared-layer structures derived from metal ions and 5-(pyridin-4-yl)isophthalic acid with or without pillars

Three new coordination polymers, [Mn(L)(H₂O)₂]₂·5H₂O (1), [Cu(L)(4,4′-bipy)]₂·H₂O (2) and [Pb(L)(4,4′-bipy)_0.5] (3) (H₂L = 5-(pyridin-4-yl)isophthalic acid; 4,4′-bipy = 4,4′-bipyridine) have been synthesized and characterized by IR, thermogravimetric analysis, X-ray powder diffraction and X-ray single crystal diffraction. Complex 1 shows a 2D layer structure which stacks with the other ones to form 1D channels to hold 1D water chains. Complex 2 displays a 3D + 3D → 3D polycatenation network. 3 shows a unique 2D + 2D → 3D polycatenation net in which each Pb center has a hemidirected coordination geometry due to the existence of “inert pair effect”. The thermal and luminescent properties of 1–3 were also examined.     

High yield expression and efficient purification of deuterated human protein galectin-2

Structural studies of biological macromolecules often require deuterated proteins, necessitating an effective bioprocessing strategy for high yield deuteration and purification. The fermentation and bioseparation studies reported here concern deuterated human protein galectin-2 mutant C57M (hGal-2), a protein showing potential for therapeutic applications. Using the vector pET-28a and a defined D₂O based minimal medium with glycerol as the sole carbon source and kanamycin for selection, we have demonstrated that a high density of Escherichia coli expressing deuterated protein at a bench bioreactor scale (7 L) can be achieved, with due attention to prevention of oxygen limitation. Yields achieved were 58 g/L biomass (wet weight) containing 0.7 g/L hGal-2. Affinity chromatography and ion-exchange chromatography were combined to achieve high purity as well as removal of hGal-2 aggregates, giving an overall yield of 1200 mg deuterated hGal-2. The deuterated hGal-2 was characterized and compared with the non-deuterated protein by size exclusion chromatography (SEC), HPLC, N-terminal sequencing, mass spectrometry (MS) and a dot blot immunoassay, showing that deuteration and subsequent purification did not impact the lactose binding and antibody recognition abilities of hGal-2. MS for both intact and trypsin-digested hGal-2 demonstrated that the extent of labeling of non-exchangeable hydrogen atoms by deuterium was (66 ± 1)%, which provides sufficient contrast variation for structural studies using small angle neutron scattering. The fermentation and bioseparation method established in this work can be applied to process other deuterated proteins with high yield and purity, opening the way to advanced structural studies.     

Preparation and characterization of lead-free (K0.47Na0.51Li0.02)(Nb0.8Ta0.2)O3 piezoceramic/epoxy composites with 0–3 connectivity

Lead-free (K_0.47Na_0.51Li_0.02)(Nb_0.8Ta_0.2)O₃ (KNLNT) piezoceramic/epoxy composites with 0–3 connectivity were prepared using cold-pressing. The dielectric and piezoelectric properties of the composites were examined as a function of mean particle size (D) within the range of 27–174 μm at a fixed ceramic content of 85 vol%. The dielectric constant increased with D by the combined effects of increased connectivity and decreased surface-to-volume ratio of ceramics. When D = 125 μm, the piezoelectric constant showed a highest value of 44 pC/N that is much greater than those of previous reports on lead-free piezoelectric 0–3 ceramic/polymer composites.     

Modeling and optimizing of electrochemical oxidation of C.I. Reactive Orange 7 on the Ti/Sb–SnO2 as anode via response surface methodology

The decolorization and degradation of an organic dye, Reactive Orange 7 (RO7) in aqueous media by electrochemical oxidation process using Ti/Sb–SnO₂ electrode as anode was modeled and optimized using response surface methodology (RSM) based on central composite design (CCD). The anode electrode was prepared using dip-coating and thermal decomposition method. Accordingly reduced quadratic model was developed to give the substrate color removal efficiency percentage as function of effective parameters such as: initial dye concentration, pH of the solution, electrolyte concentration and current density. The fit of the model is checked by the determination coefficient (R²). In this case, the value of the determination coefficient (R² = 0.9949) is indicated. Maximum color removal efficiency was achieved at the obtained conditions of: pH = 4, concentration of electrolyte = 3.5 g/L and current density = 19 mA/cm². Dye removal rate increased by increasing the concentration of electrolyte, lowering pH and increasing the current density. In optimum conditions, decolorization was obtained completely after 5 min; and the removal of chemical oxygen demand (COD) was reduced to 70.3% after 90 min.     

The production of amorphous regions in carbon nanotubes by 140 keV He ion irradiation

The production of amorphous regions in carbon nanotubes irradiated with 140 keV He ions was studied using Raman spectroscopy and transmission electron microscopy (TEM). Intensity ratios of Raman D to G bands (I_D/I_G) initially increase and then decrease as a function of ion fluence at all investigated irradiation temperatures (room temperature, 200 and 400 °C). The critical ion fluences corresponding to the maximum in I_D/I_G ratios increase with increasing irradiation temperature because of the enhanced defect annealing. The displacement per atom (dpa) values, consistent with a maximum in I_D/I_G ratios, are determined to be 0.15 dpa at room temperature and 200 °C, and 0.3 dpa at 400 °C. TEM examination of all irradiated specimens supports Raman results indicating that the maximum in I_D/I_G correlates to the formation of amorphous regions. The study shows that after formation of amorphous regions at high fluences, I_D/I_G ratio can be no longer used to measure amorphous/graphitic content in CNTs.     

Extraction of sunflower oil with supercritical CO2: Experiments and modeling

Extraction of sunflower oil from sunflower seeds (Heliantus annuus L.) using supercritical CO₂ was studied. The shrinking core model was applied to the modeling of the packed-bed extraction process. The experimental data were obtained for extraction conducted at the pressures of 20, 30, 40, 50 and 60 MPa; the temperatures of 313, 333 and 353 K, the CO₂ flow rates of 1–4, and 6 cm³ CO₂ min⁻¹; the mean particle diameters of 0.23, 0.55, 1.09, 2.18 mm. The supercritical CO₂ extraction process was modeled by a quasi steady state model as a function of extraction time, pressure, temperature, CO₂ flow rate, and particle diameter. The supercritical CO₂ extraction process. The intraparticle diffusion coefficient (effective diffusivity) D_e was used as adjustable parameter. The model using the best fit of D_e was correlated the data satisfactorily.     

Screening of twelve Clostridium botulinum (group I) spores for high-pressure resistance at elevated-temperatures

Twelve strains of Clostridium botulinum group I spores, suspended in phosphate buffer (0.1 M) at approximately 10⁷ CFU/ml concentration, were subjected to high pressure treatments (800 and 900 MPa; 0.5–15 min) at elevated temperatures (90 and 100 °C). The treatments were chosen to have a range of pressure/temperature severity to be able to discriminate the spore strains for their pressure resistance. An insulated test chamber was used to achieve temperature stability during treatment. Preliminary tests showed the need for an 8 day anaerobic incubation for enumeration. Results showed that strains PA9508B, HO9504A and CK2-A had higher pressure resistance than others among the 12 strains studied. Strain 62A was least resistant and completely inactivated by the treatment. Estimated D values of the more resistant strains were in the 0.66–1.8 min range at 900 MPa at 100 °C treatment. The temperature sensitivity parameter (Z_P value) in the 800–900 MPa pressure range varied between 10 and 16 °C, and pressure sensitivity parameter (Z_T value) in the 90–100 °C temperature range varied between 340 and 760 MPa. The most pressure resistant strain was PA9508B with an estimated Z_P value of 16.0 °C and Z_T value of 470 MPa. Since the pathogenic strains of C. botulinum have different pressure resistance, the most resistance strain should be selected as the basis for process establishment.     

Micromechanisms of solid electrolyte interphase formation on electrochemically cycled graphite electrodes in lithium-ion cells

The microstructural and compositional changes that occurred in the solid electrolyte interphase (SEI) formed on graphite electrodes subjected to voltammetry tests (vs. Li/Li⁺) at different voltage scan rates were investigated. The microstructure of the SEI layer, characterized using high-resolution transmission electron microscopy, consisted of an amorphous structure incorporating crystalline domains of ～5–20 nm in size. Evidence of lithium compounds, namely Li₂CO₃ and Li₂O₂, and nano-sized graphite fragments was found within these crystalline domains. The morphology and thickness of the SEI depended on the applied voltage scan rate (dV/dt). The variations in the Li⁺ diffusion coefficient (D_Li⁺) at the electrode/electrolyte interface during the SEI formation process were measured and two regimes were identified depending on the scan rate; for dV/dt ≥ 3.00 mV s^?1, D_Li⁺ was 3.13 × 10^?8 cm² s^?1. At lower scan rates where D_Li⁺ was low, 0.57 × 10^?8 cm² s^?1, a uniform and continuous SEI layer with a tubular morphology was formed whereas at high dV/dt, the SEI formed had a columnar morphology and did not provide a uniform coverage.     

Relationship between mechanical properties and coefficient of friction of sputtered a-C/Cu composite thin films

The article reports on properties of a-C films containing different amount of Cu. Films were sputtered by unbalanced magnetron from a graphite target with Cu fixing ring in argon under different deposition conditions. Relationships between the structure, mechanical properties, macrostress σ and coefficient of friction (CoF) μ of a-C/Cu films sputtered on Si substrates were investigated in detail. Besides, a special attention was concentrated on investigation of the effect of a deposition rate a_D of the a-C/Cu film on its hardness H and macrostress σ. Four main issues were found: (1) the addition of Cu into a-C film strongly influences its structure and mechanical properties, i.e. the hardness H, effective Young's modulus E⁎ macrostress σ and CoF, and makes it possible to form electrically conductive films; here E⁎ =  E / (1 − ν²), E is the Young's modulus, and ν is the Poisson's ratio, (2) the hardness H and compressive macrostress σ of the a-C/Cu film decrease with increasing a_D due to decreasing of total energy E_T delivered to the film during its growth, (3) hard a-C/Cu films with low value of CoF (μ ≈ 0.1) can be sputtered at high deposition rates a_D ranging from ~ 10 to ~ 80 nm/min, and (4) CoF decreases with increasing (i) hardness H and (ii) resistance of film to plastic deformation characterized by the ratio H³/E⁎² but only in the case when compressive macrostress σ is low.     

Influence of heating and laser irradiation on the Raman D band in single-wall carbon nanotubes

Fine structure of the Raman D band in single-wall carbon nanotubes (SWNTs) was investigated by heating and laser irradiation. It is shown that the D band is composed of three components at ～ 1313, 1340, and 1355 cm^? 1, denoted by D₁, D₂, and D₃, respectively. The D₁ and D₂ intensities significantly increase with laser irradiation in air and vacuum, respectively. The D₃ intensity drastically increases with heating in air. From these results, it is suggested that the fine structure of the D band is attributed to different kinds of defects introduced in SWNTs.     

Biodegradable latexes from animal-derived waste: Biosynthesis and characterization of mcl-PHA accumulated by Ps. citronellolis

Backgroundmcl-PHA biosynthesis by Pseudomonas citronellolis from tallow-based biodiesel as inexpensive carbon feed stock was accomplished. Fermentation protocols, kinetic analysis, an efficient product recovery strategy, and a detailed product characterization are presented.ResultsA maximum specific growth rate, μ_max. of 0.10 and 0.08 h^?1, respectively, was achieved in two different fermentation set-ups. Volumetric productivity for mcl-PHA amounted to 0.036 g/L h and 0.050 g/L h, final intracellular PHA contents calculated from the sum of active biomass and PHA to 20.1 and 26.6 wt.%, respectively. GC-FID analysis showed that the obtained biopolyester predominantly consists of 3-hydroxyoctanoate and 3-hydroxydecanoate, and, to a minor extent, 3-hydroxydodecanoate, 3-hydroxynonanoate, 3-hydroxyhexanoate, and 3-hydroxyheptanoate monomers. This was confirmed by ¹H- and ¹³C NMR, also evidencing the occurrence of low quantities of unsaturated and 3-hydroxyvalerate building blocks. High purity of the recovered materials was proofed by elemental analysis. Regarding the results from thermogravimetric analysis, differential scanning calorimetry and molecular mass determination, results were in a range typical for this type of PHA (1st fermentation: decomposition temperature T_d = 296 °C, peak of melting range T_m = 48.6 °C; glass transition temperature T_g = ?46.9 °C, degree of crystallinity X_c = 12.3%, M_w = 66,000, M_n = 35,000, dispersity index P_i = 1.9; 2nd fermentation: T_d = 295 °C, T_m = 53.6 °C, T_g = -43.5 °C, X_c = 10.4%, M_w = 78,000, M_n = 196,000, P_i = 2.5).