Rheological properties of milk gels formed by a combination of rennet and glucono-delta-lactone.

The effects of heat treatment of milk, and a range of rennet and glucono-delta-lactone (GDL) concentrations on the rheological properties, at small and large deformation, of milk gels were investigated. Gels were made from reconstituted skim milk at 30 degrees C, with two levels each of rennet and GDL. Together with controls this gave a total of sixteen gelation conditions, eight for unheated and eight for heated milk. Acid gels made from unheated milks had low storage moduli (G') of < 20 Pa. Heating milks at 80 degrees C for 30 min resulted in a large increase in the G' value of acid gels. Rennet-induced gels made from unheated milk had G' values in the range approximately 80-190 Pa. However, heat treatment severely impaired rennet coagulation: no gel was formed at low rennet levels and only a very weak gel was formed at high levels. In gels made with a combination of rennet and GDL unusual rheological behaviour was observed. After gelation, G' initially increased rapidly but then remained steady or even decreased, and at long ageing times G' values increased moderately or remained low. The loss tangent (tan delta) of acid gels made from heated milk increased after gelation to attain a maximum at pH approximately 5.1 but no maximum was observed in gels made from unheated milk. Gels made by a combination of rennet and GDL also exhibited a maximum in tan delta, indicating increased relaxation behaviour of the protein-protein bonds. We suggest that this maximum in tan delta was caused by a loosening of the intermolecular forces in casein particles caused by solubilization of colloidal calcium phosphate. We also suggest that in combination gels made from unheated milk a low value for the fracture stress and a high tan delta during gelation indicated an increased susceptibility of the network to excessive large scale rearrangements. In contrast. combination gels made from heated milk formed firmer gels crosslinked by denatured whey proteins and underwent fewer large scale rearrangements.

Influence of calcium, magnesium, or potassium ions on the formation and stability of emulsions prepared using highly hydrolyzed whey proteins.

Oil-in-water emulsions (4 wt % soy oil) containing 4 wt % whey protein hydrolysate (WPH) (27% degree of hydrolysis) and different levels of calcium, magnesium, or potassium chloride were prepared in a two-stage homogenizer. Other emulsions containing 4 wt % WPH but including 0.35 wt % hydroxylated lecithin and different levels of the above minerals were similarly prepared. The formation and stability of these emulsions were determined by measuring oil droplet size distributions using laser light scattering and by confocal scanning laser microscopy and a gravity creaming test. Both lecithin-free and lecithin-containing emulsions showed no change in droplet size distributions with increasing concentration of potassium in the range 0-37.5 mM. In contrast, the diameter of emulsion droplets increased with increasing calcium or magnesium concentration >12.5 mM. Emulsions containing hydroxylated lecithin were more sensitive to the addition of calcium or magnesium than the lecithin-free emulsions. Storage of emulsions at 20 degrees C for 24 h further increased the diameter of droplets and resulted in extensive creaming in emulsions containing >25 mM calcium or magnesium. It appears that both flocculation and coalescence processes were involved in the destabilization of emulsions induced by the addition of divalent cations.

Characterization of commercial and experimental sodium caseinates by multiangle laser light scattering and size-exclusion chromatography.

A range of sodium caseinate samples were characterized by a multiangle laser light scattering (MALLS) system or by the use of MALLS as an on-line detector with size-exclusion chromatography (SEC). Sodium caseinate solutions, analyzed using a MALLS system alone, gave weight-average molar mass (M(w)) values in the range 1200-4700 kDa and z-average root-mean-square radius (R(g)) values ranged from approximately 50 to 120 nm. When these solutions were ultracentrifuged at 90000g for 1 h, a cloudy top layer was formed; the subnatant was carefully removed and analyzed by SEC-MALLS. The M(w) values were found to be in the range approximately 30-575 kDa, and R(g) values ranged from approximately 22 to 49 nm. During SEC, the MALLS system detected some very large-sized material that eluted close to the void volume; this material was hardly detected by the concentration detectors, i.e., ultra-violet (UV) and differential refractive index (DRI). The intensity of the light scattering (LS) signal from this very large sized material was greatly reduced in the subnatant. SEC of sodium caseinate samples revealed two main peaks with M(w) of approximately 420-750 kDa and 39-69 kDa, respectively. The R(g) values were very large for a protein molecule, and initial calculations suggested that the shape of caseinate molecules was likely to be highly elongated.

Desorption of Trichoderma reesei cellulase from cellulose by a range of desorbents.

The desorption of Trichoderma reesei cellulase from Avicel by a wide range of desorbents was measured. Emphasis was placed on desorption at alkaline pH. A maximum desorption of 65-68% Avicelase activity was achieved by contact with NaOH, pH 10.0, at 40 degrees C for 5 min in the presence of 0.005% Triton X-100 or Tween 80. The design of a suitable desorption process using these conditions is discussed. Glycerol was also effective as a desorbent either alone or in combination with alkali and detergent. However, relatively high concentrations of glycerol were needed and the maximum desorption achieved, 68%, was not significantly greater than that with only alkali and detergent.

Centrifugal dewatering of acid casein curd: effect of casein manufacturing and centrifugation variables on curd compression in a laboratory centrifuge.

Data relevant to curd compression in a horizontal, solid bowl decanter centrifuge have been obtained by studying the dewatering of acid casein curd in a batch laboratory centrifuge. Analysis of curd compression under centrifugal force predicts a moisture content gradient in the dewatered curd from a maximum at the curd-liquid interface to a minimum at the centrifuge bowl wall. This moisture content gradient was also measured experimentally, and its practical implications are discussed. Increases in centrifugal force, centrifugation time, and centrifugation temperature all caused a marked de crease in dewatered curd moisture content, whereas in creases in precipitation pH and maximum washing temperature caused a smaller decrease in dewatered curd moisture content.

Solubilisation of bovine rumen and decolorisation of bovine blood by enzymic hydrolysis with alcalase.

Enzymic hydrolysis has potential for upgrading a range of waste materials from the meat industry. The effects of the reaction variables pH, time, temperature, enzyme concentration and rumen concentration on the solubilisation of rumen by Alcalase have been determined. Reaction progress curves showed rapid initial rates followed by a gradual decline in rate. The optimum reaction pH was 8·5. Optimum reaction temperatures for hydrolysis and solubilisation were about 55°C and 60°C, respectively, for a 3-h reaction. Solubilisation and degree of hydrolysis both increased with increasing enzyme concentration and decreased with increasing substrate concentration.

Effect of microorganisms on rate of liquid extraction of ethanol from fermentation broths.

Liquid extraction is one means of removing metabolic products continuously during a fermentation and so reducing product inhibition. It is known that microbial organisms are attracted to liquid-liquid interfaces, and it is important for the design of extraction systems to establish if this has a detrimental effect on the rate of extraction. The extraction of ethanol from aqueous suspensions of yeast (Saccharomyces cerevisiae) using n- decanol is described in this paper. It was found that the presence of the yeast cells severely reduced the rate of ethanol extraction. The overall mass transfer coefficient was reduced from 5.0 x 10(-6) to 0.7 x 10(-6) m/s. This reduced overall mass transfer coefficient was unaffected by yeast concentration in the range 0.1-20 kg/m(3). The results are consistent with the yeast cells adsorbing to the interface in closely packed layers and preventing mass transfer by simply reducing the available interfacial area. Optical microscope observations confirmed that a yeast layer several cell diameters thick rapidly built up at the interface when a small decanol droplet was added to a yeast suspension.

Kinetics of the hydrolysis of lean meat protein by alcalase: Derivation of two alternative rate equations and their fit to experimental data.

Two rate equations have been developed to model the hydrolysis of ground lean meat protein by Alcalase. The first equation was based on classical Michaelis-Menten kinetics and the second on the adsorption of enzyme to the protein prior to reaction. It was assumed that this adsorption could be modelled by a Langmuir-type adsorption isotherm. Each equation considered the enzyme to be competitively inhibited by reaction product, and considered enzyme inactivation to be first order. Both rate equations have been fitted to experimental data obtained from the hydrolysis of meat protein by Alcalase. Initial rate data indicated that the adsorption model was more appropriate. However, both equations gave satisfactory fits to 11 reaction progress curves determined over a wide range of enzyme and substrate concentrations.

Hydrolysis of the sarcoplasmic, myofibrillar and connective tissue proteins of lean beef by alcalase and its relationship to whole meat hydrolysis.

The enzymic hydrolysis and solubilisation of the three meat protein fractions have been investigated in order to elucidate some aspects of the hydrolysis of whole beef protein. Much of the water-soluble sarcoplasmic protein fraction was insolubilised on heating for 25 min at 60°C. While most of this insoluble protein was easily resolubilised enzymically, some of it remained insoluble after 5 hours' hydrolysis. Solubilisation of connective tissue by Alcalase after a 3 h reaction increased markedly from 23% at 55°C to 99% at 60°C. This explains the high optimum temperature for solubilisation of whole beef by Alcalase. A significant portion of myofibrillar tissue remained insoluble after 3 hours' reaction. indicating that the majority of the insoluble solids remaining after whole meat hydrolysis at 60°C derived from myofibrillar tissue. There was good agreement between an experimental reaction progress curve for whole beef hydrolysis and one estimated from progress curves for hydrolysis of the three meat protein fractions.

Effects of reaction variables on the hydrolysis of lean beef tissue by alcalase.

The effects of the reaction variables, pH, time, temperature, enzyme: meat ratio and meat concentration, on the hydrolysis and solubilisation of lean meat protein by Alcalase have been determined. Experiments were performed with both dilute meat suspensions and undiluted ground meat. Reaction progress curves were similar to those for the enzymic hydrolysis of other proteins, with rapid initial reaction rates followed by a gradual decline in rate. The optimum reaction pH was 8·5. Optimum temperatures were found to be 55°C for the hydrolysis reaction and 60°C for meat solubilisation. Both solubilisation and degree of hydrolysis increased with increasing enzyme concentration. At an enzyme: meat ratio of 80 mg Alcalase 0·6L per gram of dry meat, a solubilisation of 94% and a degree of hydrolysis of 18% were achieved after a 3-h reaction period. As meat concentration was increased, degree of hydrolysis decreased slightly and solubilisation decreased markedly.

New concepts for rapid yeast settling. I. Flocculation with an inert powder.

A novel technique for settling microorganisms has been described. The technique involves adding a dense, inert powder to a suspension of microorganisms under conditions where flocculation of the microorganism with the inert poweder occurs. The flocs formed are small and relatively dense and settle rapidly. Suspensions of Saccharomyces cerevisiae yeast have been flocculated with several different inert seed materials achieving rapid settling and separations of up to 99.9%. Nickel powder was used as a seed material for most experiments described here, and iron sand showed promise as a cheaper seed for large-scale use. The degree of flocculation and cell separation obtained depended largely on the seed concentration and the components in solution. Temperature and pH had little effect. When the method was initially applied to a practical fermentation, flocculation was poor because of inhibiting compounds in the fermentation medium, but modification of the technique produced good flocculation in the medium.

New concepts for rapid yeast settling. II. pH switching with an inert powder.

A new technique is outlined for the rapid settling of yeast cells in fermentation media. The technique involved the addition of dense, inert particles (nickel powder) to a yeast suspension (Saccharomyces cerevisiae) at pH 4.5 and a rapid change of pH to 8.0-9.0. When the pH was changed large flocs formed immediately and settled rapidly, leaving a clear supernatant. On returning the pH to 4.5 the flocs were destroyed. This technique gave larger flocs and higher settling rates than the constant pH method, and much lower nickel/yeast ratios were required. Good flocculation also occurred in a fermentation medium. The technique was used to recycle yeast cells to a semicontinuous ethanol fermentation. Application of the technique to this and similar systems is discussed. The factors affecting yeast/inert powder flocculation are also discussed and a model is proposed to explain the observed experimental behavior for flocculation with a rapid change in pH.

The tensile properties of frozen and thawed lean beef.

The tensile properties of frozen beef are relevant to many size reduction operations in frozen meat processing. The tensile properties of lean beef aligned both along, and perpendicular to, the fibre direction have been studied over the temperature range +10°C to -30°C and over a range of strain rates. Strain rate had an effect on tensile strength and other tensile parameters for frozen beef with either fibre orientation. Two distinct types of behaviour were observed. At the higher temperatures (e.g. -5°C and -10°C for beef oriented perpendicular to the fibre direction) there was a rise in tensile strength with increase in strain rate. This viscoelastic fracture behaviour can be attributed to the viscoelastic character of meat proteins and also to the plasticity of ice. At the lower temperatures (e.g. -15°C to -30°C for beef oriented perpendicular to the fibre direction) there was a decrease in tensile strength with increase in strain rate. This fracture behaviour can be attributed to the brittleness of ice. Temperature had a marked effect on all tensile parameters. The maximum tensile strength at most strain rates occurred in the temperature range -10°C to -20°C, and the strength was lower at -30°C than at -20°C.

Nonporous magnetic materials as enzyme supports: studies with immobilized chymotrypsin.

Chymotrypsin has been immobilized to several nonporous magnetic materials. Nickel particles were considered to be most suitable as immobilized enzyme supports. Chymotrypsin immobilized to nonporous magnetic supports was not fouled significantly by either whole milk or clarified yeast homogenate. AE-cellulose-chymotrypsin was rapidly fouled by both these materials and chymotrypsin immobilized to acrylic-based ion exchangers was slowly fouled. Immobilized enzyme activity was found to be inversely proportional to particle diameter for nonporous rock magnetic particles. Immobilization by adsorption and then glutaraldehyde crosslinking was used to produce controlled amounts of chymotrypsin on the particles. Esterolytic activity increased with enzyme loading but caseinolytic activity did not increase. Chymotrypsin is inhibited by metal ions from the magnetic supports. It is partially protected by use of a preliminary protein coating and may be reactivated by incubation with EDTA or BSA.