Crystallization kinetics of amorphous lactose, whey-permeate and whey powders

Amorphous lactose, whey-permeate and whey powders have been converted to their crystalline forms by exposure to air at various temperatures and relative humidities. The total time required for sorption, induction and crystallization of these powders was observed by following the time-dependent mass change of the powders during treatment. These experiments have shown that higher temperatures and relative humidities lead to shorter crystallization times. Lactose crystallizes within 1 min at an air temperature of 100 degrees C and relative air humidity of 80%, whereas whey-permeate and whey powders requires up to 5 min at the same set of conditions. Thus, as previously described, the presence of proteins and salts in the whey-permeate and whey powders reduces the crystallization rate. The rate constants and activation energies have been determined over a range of temperatures and humidities to enable the calculation of crystallization times for the design of an industrial process that crystallizes whey and whey-permeate powders. Finally, the crystallization rates found in this work are sufficiently fast to be applicable in an industrial process that crystallizes whey and whey-permeate powders.     

Ethanol production from cheese whey permeate by Kluyveromyces marxianus UFV-3: A flux analysis of oxido-reductive metabolism as a function of lactose concentration and oxygen levels

In order to investigate the effect of lactose concentration and oxygen level on the growth and metabolism of Kluyveromyces marxianus UFV-3 in cheese whey permeate, batch cultures were conducted under aerobic, hypoxic, and anoxic conditions, with lactose at initial concentration ranging from 1 to 240 g L⁻¹. The increase in lactose concentration increased ethanol yield and ethanol volumetric productivity, and has reduced cell yield. When lactose concentration was equal or above 50 g L⁻¹ and the oxygen levels were low, the ethanol yield was close to its theoretical value. Maximum ethanol concentrations attained in this study were 76 and 80 g L⁻¹ in hipoxia and anoxia, respectively. The lactose consumption rate in anoxia was greater than in aerobiosis and hipoxia. However, under anoxia, the lactose consumption rate of K. marxianus followed a saturation kinetics, which was not observed in hypoxia and aerobiosis. All oxygen levels investigated, showed a tendency for saturation of the ethanol production rate above 65 g L⁻¹ lactose. Ethanol production rate was also higher on anoxia.     

Crystallization and X-ray diffraction of spray-dried and freeze-dried amorphous lactose

Crystallization of spray-dried and freeze-dried amorphous lactose over different relative vapor pressures (RVP) and storage times was studied. Crystallization was observed from increasing peak intensities in X-ray diffraction patterns. Lactose was crystallized in the samples stored at RVP of 44.1% and above in both types of dehydrated powders. The rate of crystallization increased with increasing RVP and storage time. Similar crystallization behavior of both spray-dried and freeze-dried lactose was observed. Lactose crystallized as alpha-lactose monohydrate, anhydrous beta-lactose, and the anhydrous form of alpha- and beta-lactose in a molar ratio of 5:3 and 4:1 in both spray-dried and freeze-dried forms. Peak intensities of X-ray diffraction patterns for anhydrous beta-lactose were decreased, and for alpha-lactose monohydrate increased with increasing storage RVP and time. The crystallization data were successfully modeled using Avrami equation at RVP of 54.5% and above. The crystallization data obtained is helpful in understanding and predicting storage stability of lactose-containing food and pharmaceutical products.     

Research perspectives and role of lactose uptake rate revealed by its study using 14C-labelled lactose in whey fermentation

The present investigation examines the effect of pH, temperature and cell concentration on lactose uptake rate, in relation with kinetics of whey fermentation using kefir and determines the optimum conditions of these parameters. Lactose uptake rate was measured by adding ¹⁴C-labelled lactose in whey. The results reveal the role of lactose uptake rate, being the main factor that affects the rate of fermentation, in contrast to the activity of the enzymes involved in lactose bioconversion process. Lactose uptake rate results discussion showed that mainly Ca²⁺ is responsible for the reduced whey fermentation rate in comparison with fermentations using synthetic media containing lactose. Likewise, the results draw up perspectives on whey fermentation research to improve whey fermentation rate. Those perspectives are research to remove Ca²⁺ from whey, the use of nano and microtubular biopolymers and promoters such as γ-alumina pellets and volcan foaming rock kissiris in order to accelerate whey fermentation.     

FTIR study of state and phase transitions of low moisture sucrose and lactose

Mid-infrared spectra of freeze-dried sucrose and lactose systems were acquired over a range of temperatures (30-200 degrees C) and water contents (0-6.3%). Starting from the glassy state, the experimental conditions were selected to cover the main thermal transitions: the glass-rubber transition, the crystallisation and, for some samples, the subsequent melting. The FTIR spectra were very sensitive to the physical state. While subtle but systematic spectral differences between the glassy and rubbery states were detectable throughout the spectrum, a very pronounced increase in spectral resolution was observed as crystallisation occurred and was followed by the expected spectral broadening during melting. The temperatures at which these changes occurred were in satisfactory agreement with the transition temperatures measured by differential scanning calorimetry (DSC). The increase in molecular mobility as a result of increasing temperature or plasticisation by water led to a significant shift of the O-H stretching band to higher wavenumbers indicating a weakening of hydrogen bonding. This shift reached a maximum as the DSC measured crystallisation temperature range was approached. As expected, the crystallisation led to a highly effective hydrogen bonding network. This was more significant for lactose than for sucrose. No significant step change in hydrogen bonding was observed at Tg. As anticipated, the temperature at which these transitions occurred decreased with increasing water content but overlapped when observed in the context of the shifted temperature (T-Tg).     

Biodegradable biocomposite films based on whey protein and zein: barrier, mechanical properties and AFM analysis

Whey and zein protein are byproducts of the food industries and have good film making properties. Single and laminated films were produced from zein protein and whey protein and their film properties were studied. Glycerol and olive oil were used as plasticizer for the single and laminated films. The laminated films exhibited higher ultimate tensile strength (UTS) than the single whey protein films (260% and 200% in the whey-zein-glycerol and whey-zein-olive oil films, respectively). The UTS of the whey protein films increased 2-3-fold after lamination. The laminated films showed higher barrier properties than the single whey protein films (180% in the whey protein-zein-glycerol films and 200% in the whey protein-zein-olive oil films in comparison to single whey protein films) and lower than the single zein films.     

X-ray diffraction analysis of dehydrated myelin

Improved X-ray diffraction data from dry nerve myelin are presented. In addition to the spacings of approx. 150 Å, 60 Å, 44 Å and 34.6 Å, which have been previously reported, we identify a 14 Å series. The data suggests that the hydrocarbon chains in the single bilayer ($\text{≈ 60}\text{A}\text{?}$) is ordered, whereas in the double bilayer ($\text{≈ 150}\text{A}\text{?}$) and in the fluid phase ($\text{≈ 44}\text{A}\text{?}$) it is disordered. It is shown that cholesterol ($\text{≈34.6}\text{A}\text{?}$) exists as a bilayer, and the 14 Å series is probably another cholesterol phase.     

An X-ray diffraction and X-ray absorption spectroscopy joint study of neuroglobin

Neuroglobin (Ngb) is a member of the globin family expressed in the vertebrate brain, involved in neuroprotection. A combined approach of X-ray diffraction (XRD) on single crystal and X-ray absorption spectroscopy (XAS) in solution, allows to determine the oxidation state and the structure of the Fe-heme both in the bis-histidine and the CO-bound (NgbCO) states. The overall data demonstrate that under X-ray the iron is photoreduced fairly rapidly, and that the previously reported X-ray structure of ferric Ngb [B. Vallone, K. Nienhaus, M. Brunori, G.U. Nienhaus, Proteins 56 (2004) 85-92] very likely refers to a photoreduced species indistinguishable from the dithionite reduced protein. Results from the XAS analysis of NgbCO in solution are in good agreement with XRD data on the crystal. However prolonged X-ray exposure at 15 K determines CO release. This preliminary result paves the way to experiments aimed at the characterization of pentacoordinate ferrous Ngb, the only species competent in binding external ligands such as O₂, CO or NO.     

Effect of Lactose and Dried Whey Supplementation on Growth Performance and Histology of the Immune System in Broilers

The objective of this study was to evaluate the effect of lactose and dried whey supplementation as dietary component on growth performance and histology of lymphoid organs and ileum in broilers. A total of 480 day-old chicks were utilised for 42 days. Animals were assigned randomly to one of three treatments: control, lactose (2.5%), and dried whey (3.85%). Body weight was greater for animals supplemented with lactose or dried whey than for those not supplemented. There were no effects of treatments on feed intake and feed efficiency. In general, the effects of lactose or dried whey supplementation on histology of lymphoid organs and ileum were variable. Plasma cell counts were lower for animals supplemented with lactose than for those supplemented with dried whey. However, the length of intestinal villi during the starter period was greater for experimental groups than for control group.     

Two-stage fermentation process for the production of calcium magnesium acetate and propionate road deicers

Calcium magnesium acetate (CMA) and propionate (CMP) are environmentally benign deicing chemicals that can replace sodium chloride that is widely used on roads and highways at present for snow and ice control to provide safe driving conditions during winter. The price of CMA from petroleum-derived acetic acid is quite expensive. Anaerobic fermentations have not proven economical due to the low acid productivity and concentrations. A novel method for the production of CMA and CMP from lactose and whey permeate via a two-stage anoxic fermentation system, with calcium hydroxide for pH control is described in this paper. A homolactic bacterium Lactobacillus plantarum is used to convert lactose to calcium magnesium lactate (CML) in the first stage, and Propionicibacterium acidipropionici P200910 is used to convert CML to CMA and CMP in the second stage. In both stages, the conversion rates were 90% (w/w). Lactic acid productivity was 2.03 g/L/h in the first stage at a dilution ratio of 0.06 h⁻¹. Propionic and acetic acid yield was 1.79 g/L/h at a dilution rate of 0.05 h⁻¹. Calcium hydroxide addition did not significantly alter the overall yield of acids in either stage. However, the ratio of concentration of propionate to acetate in the final product changed from 3.0 when NaOH is used to 2.0 when lime is applied for pH control. After separation of the biomass, the liquid with a total concentration of 48–55 g/L of CMA and CMP can be processed to obtain a solid road deicer product.     

An X-ray diffraction study of poly-L-arginine hydrochloride

An x-ray study has been made of polyarginine hydrochloride to investigate whether, like polylysine hydrochloride, it can undergo conformational changes merely from variations in the degree of hydration. X-ray powder and fiber photographs of specimens containing up to about five molecules of water per arginine residue show features characteristic of α-helical structures including a 5.4-Å layer line and a meridional 1.5-Å reflection. Increasing the water content from ¹/₂ to 6¹/₂ molecules per residue causes the a axis of the hexagonal unit cell to increase from 14.4 Å to 15.8 Å, with no appreciable change in the 27.0 Å c axis. Removal of the last half molecule of water results in a very diffuse α pattern, but on rehydration the sharp pattern reappears. Specimens containing five to twenty water molecules per residue show quite a different pattern, the spacing of which do not vary appreciably with hydration. This pattern includes a meridional 3.4-Å reflection, a feature commonly shown by β structures, and indeed all the reflections can be satisfactorily indexed in terms of a monoclinic unit cell with a = 9.26 Å, b = 22.05 Å, c = 6.76 Å, and γ = 108.9°. These dimensions are shown by models to be compatible with a β pleated-sheet structure.     

Comparison of the aggregation behavior of soy and bovine whey protein hydrolysates

Soy-derived proteins (soy protein isolate, glycinin, and β-conglycinin) and bovine whey-derived proteins (whey protein isolate, -lactalbumin, β-lactoglobulin) were hydrolyzed using subtilisin Carlsberg, chymotrypsin, trypsin, bromelain, and papain. The (in)solubility of the hydrolysates obtained was studied as a function of pH. At neutral pH, all soy-derived protein hydrolysates, particularly those from glycinin, obtained by hydrolysis with subtilisin Carlsberg, chymotrypsin, bromelain, and papain showed a stronger aggregation compared to the non-hydrolyzed ones. This increase in aggregation was not observed upon hydrolysis by trypsin. None of the whey-derived protein hydrolysates exhibited an increase in aggregation at neutral pH. The high abundance of theoretical cleavage sites in the hydrophobic regions of glycinin probably explains the stronger exposure of hydrophobic groups than for the other proteins, which is suggested to be the driving force in the aggregate formation.     

X-ray diffraction analysis of internal wool lipids

Polarised optical microscopy (POM) and X-ray diffraction techniques were applied to intercellular lipids extracted from wool to study their structural arrangement in order to determine their role in the diffusion properties of wool fibre. Intercellular wool lipids (IWL) arranged as concentrated liposomes were shown to be a good intercellular lipid model, allowing their study by X-ray diffraction techniques. The results confirm that intercellular lipids of wool fibre are organised in a lamellar structure of 5.0–8.0 nm width, termed β-layer, which had been assumed to be lipids arranged as a bilayer. Structurally, internal wool lipids are distributed at least in two domains at low temperatures: an ordered phase made up of ceramides and free fatty acids (FFA) alone, arranged in crystal orthorhombic states separately, and a liquid crystal state when mixed together. At 40 °C there is a reversible phase transition produced by the melt of the crystal orthorhombic states, whereas the liquid crystal state remains until 65 °C.     

Direct detection of lactococcal bacteriophages in cheese whey using DNA probes

Abstract We have designed a new method for the rapid detection of lactococcal phage directly in milk samples. The method was based on a dot blot analysis and did not require a biological assay with sensitive indicator strains. Culture media or whey permeate samples containing phage were spotted directly onto a nylon membrane and the phage were lysed in situ prior to hybridization. For skim milk, whole milk and whey samples, the samples were first treated with 50 mM EDTA, 20% Triton X-100 and heated at 60°C for 5 min, prior to spotting on the membrane. The detection limit was approximately between 10⁵ and 10⁷ pfu/spot. A large number of samples could be processed simultaneously and the results were obtained within 24 h.     

Neutron and X-ray diffraction structural analysis of phosphatidylinositol bilayers

Phosphatidylinositol (PI) bilayers, squeezed together by applied osmotic pressures, were studied by both neutron diffraction and X-ray diffraction. The lamellar repeat period for PI bilayers decreased from 9.5 nm at an applied pressure of 1.1.10(6) dyn/cm2 (1.1 atm) to 5.4 nm at an applied pressure of 1.6.10(7) dyn/cm2 (16 atm). Further increases in applied pressure, up to 2.7.10(9) dyn/cm2 (2700 atm) reduced the repeat period by only about 0.3 nm, to 5.1 nm. Thus, a plot of applied pressure versus repeat period shows a sharp upward break for repeat periods less than about 5.4 nm. For repeat periods of less than 5.4 nm, analysis of neutron-scattering density profiles and electron-density profiles indicates that the structure of the PI bilayers changes as the bilayers are dehydrated, even though there are only small changes in the repeat period. These structural changes are most likely due to removal of water from the headgroup regions of the bilayer. D2O/H2O exchange experiments show that, at an applied pressure of 2.8.10(7) dyn/cm2, water is located between adjacent PI headgroups in the plane of the bilayer. We conclude that, although electrostatics provide the dominant long-range repulsive interaction, hydration repulsion and steric hindrance between PI headgroups from apposing bilayers provide the major barriers for the close approach of adjacent PI bilayers for repeat periods less than 5.4 nm. This structural analysis also indicates that the phosphoinositol group extends from the plane of the bilayer into the fluid space between adjacent bilayers. This extended orientation for the headgroup is consistent with electrophoretic measurements on PI vesicles.     

An X-ray diffraction study of poly-L-homoarginine hydrochloride.

An X-ray study of the synthetic polypeptide poly(L -homoarginine hydrochloride) has been made to investigate whether, like the chemically related polypeptides poly(L -lysine hydrochloride), poly(L -arginine hydrochloride), and poly(L -ornithine hydrobromide), it can undergo conformational transitions merely from variations in its degree of hydration. X-ray photographs of powder and oriented specimens containing one to 15 molecules of water per L -homoarginine hydrochloride residue showed that this polymer forms only a β-pleated-sheet structure. The pleated sheets, formed by antiparallel polypeptide chains hydrogen-bonded to each other, are piled up along the b axis in an alternating sequence (“sandwich structure”). This structure did not appreciably change with variations of the degree of hydration, and the observed reflections at 56% relative humidity (1.8 molecules of water per residue) could be indexed satisfactorily in terms of a monoclinic unit cell, of space group P2₁, with a = 9.34 Å, b = 40.07 Å, c = 6.94 Å, and γ = 106°. These dimensions are shown by models to be compatible with the proposed structure, and the calculated density of 1.27 g/cm³ agrees well with the experimental value of 1.29 g/cm³. Removal of the last molecule of water results in a very diffuse pattern, while specimens containing 20 molecules of water per residue show only reflections due to water.     

Biotechnological approaches for the value addition of whey

Whey, the liquid remaining after milk fat and casein have been separated from whole milk, is one of the major disposal problems of the dairy industry, and demands simple and economical solutions. In view of the fast developments in biotechnological techniques, alternatives of treating whey by transforming lactose present in it to value added products have been actively explored. Whey can be used directly as a substrate for the growth of different microorganisms to obtain various products such as ethanol, single-cell protein, enzymes, lactic acid, citric acid, biogas and so on. In this review, a comprehensive and illustrative survey is made to elaborate the various biotechnological innovations/techniques applied for the effective utilization of whey for the production of different bioproducts.     

Preliminary X-ray diffraction analysis of HhaII endonuclease-DNA cocrystals

HhaII restriction endonuclease purified from an overproducing recombinant E. coli clone has been cocrystallized with a heptanucleotide duplex, d-GGAGTCC:GGACTCC. The cocrystals are monoclonic and belong to the space group C2. The unit cell dimensions are a = 199.0 +/- 1.0 A, b = 100.0 +/- 0.5 A, c = 80.3 +/- 0.4 A, and beta = 101.0 +/- 1.0 degrees. There appear to be two dimers per asymmetric unit and the crystals diffract to 4-A resolution.