Concentration of n‐3 polyunsaturated fatty acids in cholesterol‐reduced cod‐liver oil by lipases

This study was conducted to elucidate the effects of different lipases originated from Candida rugosa (CR), porcine pancreas (PP) and Aspergillus niger (AN) on the degree of hydrolysis (DH) in cholesterol‐reduced cod‐liver oil (87.5%) and evaluate the changes in n‐3 polyunsaturated fatty acid concentrations in the oil hydrolysed by the lipases. The lipase‐catalysed hydrolysis of cholesterol‐reduced cod‐liver oil was performed at 37 °C for 8 h. Among all the lipase samples studied, DH in the oil after lipase‐catalysed hydrolysis followed the decreasing order: CR lipase (70.01%) > PP lipase (26.18%) > AN lipase (18.57%). Triacylglycerol levels in the oil hydrolysed by all the lipases studied decreased, while mono‐ and diacylglycerol levels increased during lipase‐catalysed hydrolysis. The eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) concentrations in cholesterol‐reduced cod‐liver oil hydrolysed by the CR lipase were remarkably higher than those by the PP or AN lipase. Thus, it is suggested in this study that the CR lipase appears to be most suitable for producing and n‐3 polyunsaturated fatty acids including EPA and DHA concentrates from cholesterol‐reduced cod‐liver oil.     

An improved method for the measurement of 3‐monochloropropanediol esters by matrix solid‐phase dispersion supported liquid–liquid extraction

3‐Monochloropropanediol (3‐MCPD) esters are contaminants produced from the high‐temperature processing of edible oils. The accurate measurement of 3‐MCPD using an easy‐to‐follow and reliable method that uses a readily available instrument is important. Here, we report an acid transesterification heptafluorobutyrylimidazole (HFBI) derivatisation method for the accurate measurement of 3‐MCPD esters in edible oils. We developed a dispersed matrix solid‐phase supported liquid–liquid extraction (DMSP‐SLE) system to remove impurities. Both the transesterification and DMSP‐SLE conditions were optimised. A good linear relationship was obtained within the range of 0.05–10 mg kg^?1 (R² ≥ 0.999) in both blank solvent and an oil sample. The limit of detection was 20.36 μg kg^?1. The average recovery of the 3‐MCPD esters spiked at 0.5, 1.0 and 2.0 mg kg^?1 into a blank oil matrix was in a range from 105.09 ± 2.77% to 120.16 ± 10.88%. The method we developed was further confirmed by performing detection on a Food Analysis Performance Assessment Scheme (FAPAS) sample.     

Response surface optimization and characteristics of Indica rice starch‐based fat substitute prepared by α‐amylase

Response surface methodology (RSM) was used to study the effect of enzyme to substrate ratio (11.8–23.6 U α‐amylase/g rice starch), hydrolysis temperature (90–100°C) and pH value (6.0–6.6) on the gel strength of rice starches‐based fat substitute using α‐amylase hydrolysis. The optimum conditions obtained from response surface analysis was 16.52 U/g enzyme dosage, 92°C hydrolysis temperature while the influence of pH was found insignificant in the range tested. Under these optimum conditions, the gel strength of this fat substitute was 113 g/cm², very close to the gel strength of butter of 114 g/cm², while the solubility of the substitute was 1.33 ± 0.01% and the swelling power 4.85 ± 0.02. There were no observable differences in the granular size distribution between the untreated rice starch and the hydrolyzed rice starch. Rheological properties of this rice starch‐based fat substitute implied that it is easier for the substitute to form three‐dimensional networks under 34°C.     

Improving antioxidant activities of whey protein hydrolysates obtained by thermal preheat treatment of pepsin,trypsin, alcalase and flavourzyme

Antioxidant activity of whey protein concentrate (WPC) hydrolysates was evaluated. Hydrolysates were obtained by pepsin, trypsin, alcalase and flavourzyme enzymatic reaction and preheat treatment of 95 °C for 5 or 10 min. The degree of hydrolysis (DH) was determined by 2,4,6‐trinitrobenzene sulphonic acid method, and antioxidant properties were determined by three spectrophotometric methods: ferricyanide method, ferric reducing/antioxidant power assay and diphenyl‐picryl hydrazinyl radical‐scavenging activity. For all the enzymes, briefly preheat treatment (95 °C/5 min) increased DH of WPC. Alcalase hydrolysates showed the highest antioxidant activity by three methods. The changes in antioxidant activity was coincidental with the changes in DH (R² = 0.988). Hydrolysates analysed by polyacrylamide gel electrophoresis and high performance liquid chromatography indicated that the α‐La was hydrolysed completely by pepsin, trypsin and alcalase and was resistant to flavourzyme to some extent; β‐lactoglobulin was only completely hydrolysed by trypsin and alcalase. Results indicated that antioxidant activity of hydrolysates was greatly related to the exposure of amino acid residues.     

Assessment of hydrolysis of cheese whey and use of hydrolysate for bioproduction of carotenoids by Sporidiobolus salmonicolor CBS 2636

BACKGROUND: The increasing industrial demand for carotenoids has led to growing interest in their bioproduction. The need to reduce production costs has encouraged the use of low‐cost agroindustrial substrates. In this context, this work studied the pretreatment of Mozzarella cheese whey and the use of the pretreated whey as fermentation medium for the bioproduction of carotenoids by Sporidiobolus salmonicolor CBS 2636. RESULTS: Bioproduction was carried out in an orbital shaker using a 10 mL L^?1 inoculum, incubation at 25 °C and a stirring rate of 180 rpm for 120 h in a non‐illuminated environment. The carotenoids were recovered using liquid N₂ combined with dimethyl sulfoxide for cell rupture and an acetone/methanol mixture (7:3 v/v) for extraction. The maximum concentration of total carotenoids obtained was 590.4 µg L^?1 in a medium with 900 g L^?1 cheese whey hydrolysate and 4 g L^?1 K₂HPO₄ at 180 rpm, 25 °C and pH 4. CONCLUSION: The use of enzyme‐hydrolysed cheese whey was more effective in carotenoid bioproduction by S. salmonicolor CBS 2636 than the use of acid‐hydrolysed cheese whey. The concentration of total carotenoids obtained with the enzymatic hydrolysate was six times higher than that obtained with the acid hydrolysate. Copyright © 2009 Society of Chemical Industry     

Antioxidant function of tea dregs protein hydrolysates in liposome–meat system and its possible action mechanism

Tea dregs possess abundant proteins, and the objective of this study was to investigate the antioxidant activity of tea dregs protein hydrolysate with limited hydrolysis by protamex and its possible action mechanism. Tea dregs protein was hydrolysed by alcalase, protamex or neutrase. The hydrolysis condition was optimised, and the hydrolysate was characterised for 1,1‐diphenyl‐2‐picryl hydrazyl (DPPH) radical‐scavenging activity, hydroxyl radical‐scavenging activity and antioxidant activity in linoleic acid (LA) system and in chicken products. Tea dregs protein hydrolysate (TDPH) was formulated (0.1%, 0.5%, 1.0%, w/w) into chicken products to determine in situ antioxidant efficacy. Thiobarbituric acid‐reactive substances (TBARS) and peroxide value (POV) formed in chicken products during storage (4 °C, 0–7 days) were analysed. Results showed that the optimum hydrolysis condition was at 50 °C, pH 7.0 for 20 min, and the concentration of tea dregs protein was 1.5%; ratio of protamex to substrate was 6000 U g^?1. The radical‐scavenging ratio of TDPH to 1,1‐diphenyl‐2‐picryl hydrazyl (DPPH) was 90.30% at the concentration of 0.1 mg mL^?1 and that to hydroxyl radical was 65.18% at the concentration of 1.0 mg mL^?1. Moreover, it also showed strong antioxidant activity both in linoleic acid (LA) system and in chicken products. The molecular weight distribution of tea dregs hydrolysates was determined by nanofiltration tubular membrane, and the protein hydrolysates with molecular weight above 8000 Da had more effective antioxidant activity. The radical‐scavenging activities to DPPH and hydroxyl radical were 85.72% at 0.1 mg mL^?1 and 71.52% at 1.0 mg mL^?1, respectively. These findings suggest that the enzymatic hydrolysate of tea dregs protein probably possesses the specific peptides/amino acids which could stabilise or terminate the radicals through donating hydrogen. In addition, the hydrolysate could form a physical barrier around the fat droplets.     

Characteristics of immobilised β‐glucosidase and its effect on bound volatile compounds in orange juice

In this study, bound volatile compounds were isolated and extracted with Amberlite XAD‐2 resin and then hydrolysed by free or immobilised β‐glucosidase. The released bound volatiles were analysed by GC‐MS. In addition, the optimisation of immobilisation method on sodium alginate and the characteristics of immobilised β‐glucosidase were studied. The results showed that crosslinking‐entrapment was the best method. The optimal conditions of this method were as follows: sodium alginate concentration 3.5%, glutaraldehyde concentration 1%, crosslinking time 3 h, immobilisation time 2 h and CaCl₂ concentration 3%. The optimum temperature for β‐glucosidase (65 °C) was decreased by 10 °C after immobilisation, while the optimum pH values for free and immobilised β‐glucosidase were both at pH 5.0. The K_m values of free and immobilised β‐glucosidase were 14.89 and 0.59 m , respectively. In total, thirteen and six bound volatile compounds were detected in orange juice hydrolysed by free and immobilised β‐glucosidase, including benzenic compounds, terpenic compounds, hydroxy esters, C₁₃‐norisoprenoids and alcohols.     

Prospects for the utilization of the endogenous enzymes in sorghum malt in the hydrolysis of starch: Case study with utilization of breadfruit starch for ethanol production

Hydrolysis of breadfruit flour by acid and enzymes of sorghum malt were investigated. Mash (malt to breadfruit flour) ratios of 2: 3 and 1: 1 were used. The optimum mashing profile of 20 minutes at 64°C followed by 30 minutes at 72°C was established. The addition of an exogenous enzyme improved the wort properties. Kinetic study of the enzymatic hydrolysis revealed non‐adherence with either the simple Michaelis‐Menten Kinetics or one with product inhibition. Anaerobic fermentation of the hydrolysate by Sacchammyces cerevisiae at pH 4.7 and temperature of 30°C, was carried out with and without mineral and nitrogen sources supplementation of the wort. The results showed that the enzymes in sorghum malt could be used, with some external enzyme supplementation, to hydrolyse breadfruit starch to yield appreciable amounts of sugar which may be used as food sweetener, substrate for single cell protein production, and in the production of beverages. This is particularly relevant to operations in developing countries.     

Toxicological assessment of 3-chloropropane-1,2-diol and glycidol fatty acid esters in food

Fatty acid esters of 3‐chloropropane‐1,2‐diol (3‐MCPD) and glycidol are a newly identified class of food process contaminants. They are widespread in refined vegetable oils and fats and have been detected in vegetable fat‐containing products, including infant formulas. There are no toxicological data available yet on the 3‐MCPD and glycidol esters, and the primary toxicological concern is based on the potential release of 3‐MCPD or glycidol from the parent esters by lipase‐catalyzed hydrolysis in the gastrointestinal tract. Although 3‐MCPD is assessed as a nongenotoxic carcinogen with a tolerable daily intake (TDI) of 2 μg/kg body weight (bw), glycidol is a known genotoxic carcinogen, which induces tumors in numerous organs of rodents. The initial exposure estimates, conducted by Federal Institute for Risk Assessment (BfR) under the assumption that 100% of the 3‐MPCD and glycidol are released from their esters, revealed especially that infants being fed commercial infant formula could ingest harmful amounts of 3‐MCPD and glycidol. However, the real oral bioavailability may be lower. As this gives rise for toxicological concern, the currently available toxicological data of 3‐MCPD and glycidol and their esters are summarized in this review and discussed with regard to data gaps and further research needs.     

Optimization of Fermentation Parameters for the Production of Concentrated Starters from Nonbitter Streptococcus lacti INIA 12

Streptococcus lactis INIA 12, a selected nonbitter strain, reached its maximum counts and its highest lactic acid production in a culture medium containing 125 g/L skim milk powder, fortified with 5 g/L yeast extract and digested with 10 ppm papain for 20 min at 65°C. Lactose added to the medium did not enhance growth rate or biomass production. A growth temperature of 32°C and the maintenance of pH at 6.80, with 10N NaOH as the neutralizer, were the optimum fermentation parameters in batch cultures. In ten 40-L fermentations carried out at 32°C and pH 6.80, with a 5% inoculum, a 0.2 kg/cm² nitrogen head space pressure and a stirring rate of 80 rpm, maximum counts of S. lactis (10¹⁰ CFU/mL) were reached after incubation for 6 hr at 32°C and pH 6.80.     

3‐Chloropropane‐1,2‐diol (3‐MCPD) in Soy Sauce: A Review on the Formation,Reduction, and Detection of This Potential Carcinogen

Soy sauce, a dark‐colored seasoning, is added to enhance the sensory properties of foods. Soy sauce can be consumed as a condiment or added during the preparation of food. There are 3 types of soy sauce: fermented, acid‐hydrolyzed vegetable protein (acid‐ HVP), and mixtures of these. 3‐Chloropropane‐1,2‐diol (3‐MCPD) is a heat‐produced contaminants formed during the preparation of soy sauce and was found to be a by‐product of acid‐HVP‐produced soy sauce in 1978. 3‐MCPD has been reported to be carcinogenic, nephrotoxic, and reproductively toxic in laboratory animal testing and has been registered as a chemosterilant for rodent control. 3‐MCPD is classified as a possible carcinogenic compound, and the maximum tolerated limit in food has been established at both national and international levels. The purpose of this review is to provide an overview on the detection of 3‐MCPD in soy sauce, its toxic effects, and the potential methods to reduce its concentration, especially during the production of acid‐HVP soy sauce. The methods of quantification are also critically reviewed with a focus on efficiency, suitability, and challenges encountered in analysis.     

Migration of lactic acid,lactide and oligomers from polylactide food-contact materials

Polylactide (PLA) is used for manufacturing lunch boxes and for packaging fresh food in Japan. PLA can be hydrolysed relatively easily to produce lactic acid, lactide and oligomers. Different types of PLA sheet were subjected to migration tests under various conditions and the lactic acid, lactide and oligomers contents of the migration solutions were determined using liquid chromatography/mass spectrometry (LC/MS). Furthermore, the change in molecular weight was determined by a migration test. PLA was stable at 40°C for 180 days; the total of lactic acid, lactide and oligomers migration levels were 0.28–15.00 µg cm^?2. PLA decomposed clearly at 60°C for only 10 days, the total migration levels were increased to 0.73–2840 µg cm^?2. PLA sheets with a high D-lactic acid content decomposed particularly rapidly. The amounts of alkali decomposition products, based on the conversion of lactide and oligomers to lactic acid by alkali hydrolysis, corresponded with the total migration levels.     

Prehydrolysis of xylan in culm of Sasa kurilensis with dilute sulphuric acid

To prepare a substrate for microbial conversion of xylose into xylitol, the culm of Sasa kurilensis was hydrolysed with dilute sulphuric acid. A fermentable substrate with a relatively high xylose concentration (21.9?g?L^?1) was obtained by hydrolysis with 2% sulphuric acid with a liquid to solid ratio of 10?:?1 at 121°C for 1?h. During hydrolysis at elevated temperatures, some undesirable byproducts were also generated, such as degradation products of solubilized sugars and lignin, which are potential inhibitors of microbial metabolism. These compounds were successfully removed from the hydrolysate by treatment with a commercially available activated charcoal (30?g?L^?1 dose).     

Tryptophan degradation during heat treatments: Part 2—Degradation of protein-bound tryptophan

The stability of free or protein-bound tryptophan was determined during acid (4 m methane sulfonic acid) or alkaline (4·2 m sodium hydroxide) hydrolysis at 110°C under nitrogen. The influence of time and of glucose, starch or amino acid addition was studied. It was found that hydrolysis with methane sulfonic acid caused tryptophan losses when glucose or starch was present.The thermal degradation of protein-bound tryptophan was studied using either glycyl-l-tryptophyl-glycine or casein. Analysis of tryptophan was carried out after acid or alkaline hydrolysis, through high performance liquid chromatography and UV detection.Heat treatment of tripeptide (9·4 mM, pH 8, 125°C, 3 to 48 h) in the presence of oxygen, air or nitrogen resulted in peptide hydrolysis, with the formation of the corresponding dipeptides and of free tryptophan. After acid or alkaline hydrolysis, tryptophan loss was seen to be higher when the initial heat treatment was performed in the presence of oxygen than in the presence of air.Thermal treatment of 4 or 5% w/v casein solutions (pH 7 or 8) was carried out at 125°C, under oxygen, air or nitrogen. A marked loss of tryptophan was found to occur after 24 h at 125°C when oxygen or air was present. Under nitrogen, protein-bound tryptophan was heat stable, even in the presence of glucose or starch. It is therefore unlikely that the indole ring of protein-bound tryptophan may react with reducing carbohydrates through Maillard-type reactions. Oxidative degradation of protein-bound tryptophan is also unlikely to reach any significant extent during the heat processing of foods, unless catalysts or strong oxidizing agents are present.     

Combined chemical and enzymic treatments of corn husk lignocellulosics

The potential for using corn (Zea mays L) husk residues (carbohydrates 827 g kg^?1, lignin 66 g kg^?1 DM) as a carbohydrate source for the production of soluble sugars by combined chemical pretreatment and enzymic hydrolysis was assessed. Comparative investigations of acidic and alkaline pretreatments on corn husk have shown that pentose-containing carbohydrates comprised 86–93% of the solubilised fraction. While pretreatments with 1.25 M NaOH at 25.85° C resulted in preferential extraction of hemicellulose having DP; > 12, acid pretreatments 0.51 M H₂SO₄, 0.51 M H₃PO₄ at (85° C) resulted in extensive depolymerisation of this polysaccharide. Xylose and low molecular weight carbohydrates were identified as the major products. Subsequent hydrolysis of the solubilised fraction with crude hemicellulase preparations yielded 40.90% fermentable sugars. When NaOH (0.02–1.25 M), H₂SO₄ and H₃PO₄ (0.02–0.51 M) were used as pretreatment solvents (25–85° C, 2 h), NaOH was the most effective in increasing the susceptibility of the residual husk towards enzymes, yielding 83–96% reducing sugars. This solvent solubilised up to 60.6% of the lignin and appeared to disintegrate the fibrillar structure of husk. The crystallinity of husk residues increased following the chemical pretreatments and was positively correlated with cellulose content. Enzymic hydrolysis with commercial cellulase preparations proceeded in two stages: a rapid breakdown of amorphous cellulose after which the hydrolysis rate levelled off. Similar biphasic patterns were observed for the pyrolysis temperature of cellulose. Under the most optimal conditions for husk saccharification (pretreatment with 1.25 M NaOH, 25° C, 2 h, followed by enzymic hydrolysis using a mixture of cellulase and cellobiase), 96% of the cellulose-enriched residues was hydrolysed to reducing sugars. A cellulase preparation from Trichoderma reesei exhibited substantial hemicellulolytic activity and could, therefore, be used as the sole saccharifying enzyme preparation for husk lignocellulosics.     

Production of soya milk containing low flatulence‐causing oligosaccharides in a packed bed reactor using immobilised α‐galactosidase

Peanut α‐galactosidase was immobilised in calcium alginate beads and used to hydrolyse the flatulence‐causing oligosaccharides, raffinose and stachyose, in soya milk in batch and in packed bed reactor with recycle. The immobilised enzyme exhibited a slightly lower activity than the free enzyme. The activity yield of immobilised α‐galactosidase was 75.1% and the immobilisation yield was 82.6%. Batch hydrolysis using immobilised enzyme at 55 °C resulted in 96% reduction in the oligosaccharides after 12 h. For the continuous process, a packed bed reactor with recycle was used. More than 98% of the oligosaccharides were hydrolysed after 6 h of reaction at 55 °C. The immobilised enzyme also proved to be stable up to three repeated hydrolysis reactions.     

Effects of operating parameters on acid hydrolysis of ground wheat starch: Maximization of the sugar yield by statistical experiment design

Fuels and chemicals production from carbohydrate rich waste materials has received considerable attentions in recent years. Pretreatment and hydrolysis are required stages during processing the waste materials for this purpose. Therefore, the objective of this study was to determine the optimum operating conditions in acid hydrolysis of ground wheat (GW) starch for maximum yield of total sugar (TS) formation. Hydrolysis reaction was realized by dilute sulfuric acid and heat treatment in an autoclave. Factorial experiment design was used to determine the significant variables affecting the yield of hydrolysis. The independent variables were particle size (D_p = <74 and 220 µm), acid concentration (A = 0.98–196 g/dm³), GW concentration (GW = 10–20 g/dm³), hydrolysis temperature (T = 90–130°C), and time (t = 5–60 min). The dependent variables were final TS concentration and hydrolysis yield (Y_H). The significant factors in the hydrolysis of GW were determined as GW concentration, acid (A) concentrations, and hydrolysis temperature (T). Box–Wilson statistical experimental design was used for optimization of hydrolysis conditions by considering the signficant factors. The optimal conditions, maximizing the hydrolysis yield (Y_H = 0.94) were A = 49 g/dm³ (0.5 M), T = 90°C, GW = 35 g/dm³ and t = 15 min. The most suitable A/GW ratio for the maximum hydrolysis yield of 94% was 1.3.     

Optimisation of process conditions for lactose hydrolysis in paneer whey with cold‐active β‐galactosidase from psychrophilic Thalassospira frigidphilosprofundus

The present study was conducted to analyse the physiochemical properties of Indian paneer whey. High concentration of minerals such as potassium, calcium, zinc and sodium, as NaCl, were observed which indicates the suitability of paneer whey in the preparation of beverages. A central composite rotatable design (CCRD) of response surface methodology (RSM) was employed to optimise the hydrolysis of lactose from whey using cold‐active β‐galactosidase of Thalassospira frigidphilosprofundus. Results indicated that 80% of lactose was hydrolysed at pH of 6.5 at 20 °C in 40 min in comparison with 40% at 30 °C. This emphasises the potential use of cold‐active β‐galactosidase in dairy industry.     

Fate of aflatoxin B(1) in contaminated corn gluten during acid hydrolysis

BACKGROUND: Aflatoxins are a group of mycotoxins that cause serious chronic disease outbreaks and contaminate several food products such as corn and its by‐product, corn gluten. The aim of the current study was to evaluate the effect of hydrochloric acid (HCl) on aflatoxin B₁ (AFB₁) degradation in contaminated corn gluten under different HCl concentrations, hydrolysis temperatures and hydrolysis times. RESULTS: During the wet milling process the highest AFB₁ level (45.68 µg kg^?1) (37.86%) was found in corn gluten fraction. Treatment with 1 mol L^?1 HCL at 110 °C resulted in degradation of AFB₁ by 27.6% (33.07 µg kg^?1) after 4 h and reached 42.5% (26.26 µg kg^?1) after 8 h. Increasing HCl concentration from 1 to 3 mol L^?1 HCl resulted in increased degradation of AFB₁, while complete degradation occurred in the presence of 5 mol L^?1 HCl after 4 h at 110 °C. Meanwhile, half‐life time of AFB₁ was recorded after 2 h at 100 °C and was < 2 h at 110 °C in the presence of 3 mol L^?1 HCl. CONCLUSION: It could be demonstrated that the manufacture of hydrolyzed vegetable protein is a suitable method for decontamination of aflatoxin in highly contaminated grains, especially gluten fractions. The hydrolysis reaction could be considered in terms of first‐order reaction kinetics of AFB₁ degradation. Copyright © 2010 Society of Chemical Industry