Partial Purification and Characterization of Extracellular Fructofuranosidase with Transfructosylating Activity from <Emphasis Type="Italic">Candida</Emphasis> sp.

The present work was carried out with the aim to investigate some properties of an extracellular fructofuranosidase enzyme, with high transfructosylating activity, from Candida sp. LEB-I3 (Laboratory of Bioprocess Engineering, Unicamp, Brazil). The enzyme was produced through fermentation, and after cell separation from the fermented medium, the enzyme was concentrated by ethanol precipitation and than purified by anion exchange chromatography. The enzyme exhibited both fructofuranosidase (FA) and fructosyltransferase (FTA) activities on a low and high sucrose concentration. With sucrose as the substrate, the data fitted the Michaellis–Menten model for FA, showing rather a substrate inhibitory shape for fructosyltransferase activity. The K _m and v _max values were shown to be 13.4 g L⁻¹ and 21.0 μmol mL⁻¹ min⁻¹ and 25.5 g L⁻¹ and 52.5 μmol mL⁻¹ min⁻¹ for FA and FTA activities, respectively. FTA presented an inhibitory factor K _i of 729.8 g L⁻¹. The optimum conditions for FA activity were found to be pH 3.25–3.5 and temperatures around 69 °C, while for FTA, the optimum condition were 65 °C (±2 °C) and pH 4.00 (±0.25). Both activities were very stable at temperatures below 60 °C, while for FA, the best stability occurred at pH 5.0 and for FTA at pH  4.5–5.0. Despite the strong fructofuranosidase activity, causing hydrolysis of the fructooligosaccharides (FOS), the high transfructosilating activity allows a high FOS production from sucrose (44%).     

Production of Cell Membrane-Bound α- and β-Glucosidase by Lactobacillus acidophilus

Hydrolytic enzymes, viz. α- and β-glucosidase, were produced from indigenous isolate, Lactobacillus acidophilus, isolated from fermented Eleusine coracana. Production of these enzymes was enhanced by optimizing media using one factor at a time followed by response surface methodology. The optimized media resulted in a 2.5- and 2.1-fold increase in α- and β-glucosidase production compared with their production in basal MRS medium. Localization studies indicated 80% of the total activity to be present in the cell membrane-bound fraction. Lack of sufficient release of these enzymes using various physical, chemical, and enzymatic methods confirmed their unique characteristic of being tightly cell membrane bound. Enzyme characterization revealed that both α- and β-glucosidase exhibited optimum catalytic activity at 50 °C and pH 6.0 and 5.0, respectively. K _m and V _max of α-glucosidase were 4.31 mM and 149 μmol min⁻¹ mL⁻¹ for p-nitrophenyl-α-d-glucopyranoside as substrate and 3.8 mM and 120 μmol min⁻¹ mL⁻¹ for β-glucosidase using p-nitrophenyl-β-d-glucopyranoside as the substrate.     

Properties of <Emphasis Type="Italic">Aspergillus subolivaceus</Emphasis> free and immobilized dextranase

Aspergillus subolivaceus dextranase is immobilized on several carriers by entrapment and covalent binding with cross-linking. Dextranase immobilized on BSA with a cross-linking agent shows the highest activity and considerable immobilization yield (66.7%). The optimum pH of the immobilized enzyme is shifted to pH 6.0 as compared with the free enzyme (pH 5.5). The optimum temperature of the reaction is resulted at 60 °C for both free and immobilized enzyme. Thermal and pH stability are significantly improved by the immobilization process. The calculated K _m of the immobilized dextranase (14.24 mg mL⁻¹) is higher than that of the free dextranase (11.47 mg mL⁻¹), while V _max of the immobilized enzyme (2.80 U μg protein⁻¹) is lower than that of the free dextranase (11.75 U μg protein⁻¹). The immobilized enzyme was able to retain 76% of the initial catalytic activity after 5.0 cycles.     

An approach to improve ACE-inhibitory activity of casein hydrolysates with plastein reaction catalyzed by Alcalase

The preparation method of casein hydrolysates with high ACE-inhibitory activity was studied by Alcalase-catalyzed hydrolysis coupled with plastein reaction. Casein hydrolysates with an IC₅₀ value of about 47 μg mL⁻¹ were first prepared by hydrolysis of casein with Alcalase and then modified with plastein reaction catalyzed by the same enzyme. The impacts of four reaction conditions on plastein reaction of casein hydrolysates were studied, and then optimal conditions were determined using response surface methodology with the decrease of free amino groups in the reaction mixture as response. When the concentration of casein hydrolysates was fixed at 35% by weight, the maximum decrease of free amino groups in the reaction mixture of 181.8 μmol g⁻¹ proteins was obtained. The optimum conditions for the above decrease were found to be an E/S ratio of 7.7 kU g⁻¹ proteins, reaction temperature of 42.7 °C and reaction time of 6 h. Analysis results showed that ACE-inhibitory activity of casein hydrolysates prepared could be improved significantly by plastein reaction. When casein hydrolysates were modified by plastein reaction, with a decrease of free amino groups in the mixture of about 154.7 μmol g⁻¹ proteins and 181.8 μmol g⁻¹ proteins, their IC₅₀ values could be decreased to 0.6 and 0.5 μg mL⁻¹.     

Preparation of Alcalase-catalyzed casein plasteins in the presence of proline addition and the ACE-inhibitory activity of the plasteins in vitro

Casein hydrolysates were prepared by hydrolysis of casein with alkaline protease Alcalase for 6 h and showed the highest ACE-inhibitory activity in vitro with an IC₅₀ value of 47.1 μg mL⁻¹. Casein hydrolysates prepared were subjected to Alcalase-catalyzed plastein reaction in the presence or absence of proline addition to prepare casein plasteins. Some optimal reaction conditions of plastein reaction in the presence of proline addition were studied using response surface methodology with the decrease in free amino groups in the casein plasteins as response. When the concentration of casein hydrolysates was fixed at 35% (w w⁻¹) and reaction time at 6 h, the optimal conditions were reaction temperature 48 °C, addition level of proline 0.54 mol/mol free amino groups of casein hydrolysates and addition level of Alcalase 9.5 kU g⁻¹ proteins. With these conditions, the maximal decrease in free amino groups in casein plasteins was 195.7 μmol g⁻¹ proteins. The ACE-inhibitory activities of twelve casein plasteins in vitro, prepared in the presence or absence of proline addition with different reaction extents, were evaluated and compared. The results showed that the ACE-inhibitory activity of the casein plasteins prepared in the presence of proline addition changed irregularly, different to that of the casein plasteins prepared in the absence of proline addition, and might relate to the different linking of proline to the peptides in casein hydrolysates during plastein reaction. When the casein plasteins prepared in the presence of proline addition had a decrease in free amino groups 195.7 μmol g⁻¹ proteins, the IC₅₀ value of the casein plasteins was lowered to 0.2 μg mL⁻¹.     

Large-scale production and application of leucine aminopeptidase produced by <Emphasis Type="Italic">Aspergillus oryzae</Emphasis> LL1 for hydrolysis of chicken breast meat

Leucine aminopeptidase (LAP) production by Aspergillus oryzae LL1 (LAP LL1) was scaled up by varying fermenting conditions. The maximal LAP activity for 5, 20, 250, and 2,000 L fermenters were 0.22, 0.16, 0.19, and 0.12 U mL⁻¹, respectively. LAP LL1 production coincides with increasing pH for all scale cases. A pilot plant scale hydrolysis experiment using LAP LL1 was prepared by removing the cells from 1,000 L of A. oryzae LL1 culture and concentrating the supernatant to ∼60 L, with a LAP LL1 activity yield of ∼20% and a specific LAP activity of 0.054 U mg⁻¹, comparable to Flavourzyme. Using a casein substrate, LAP LL1 specific protease activity (50.9 U mg⁻¹) is 43% higher than Flavourzyme (35.6 U mg⁻¹). We believe LAP LL1 has potential use as an industrial enzyme for food processing. Comparing the hydrolysis effects on chopped chicken breast meat (100 g), the degree of hydrolysis (D_H) of LAP LL1 (50.6%) was about twice as effective as Flavourzyme (25.6%). On a pilot-plant scale hydrolysis experiment of chopped chicken breast meat (54 kg), the D_H of LAP LL1 was 45.6%. Adding 30 μmol L⁻¹ zinc increased D_H by 33.3% for LAPs LL1 but zinc did not enhance Flavourzyme hydrolysis. Together, this study provides possibility that LAP LL1 has significant potential for application in the food industry. Shie-Jea Lin and Yi-Hong Chen have contributed equally to this work.     

Antioxidant capacity and antioxidative compounds in barley (<Emphasis Type="Italic">Hordeum</Emphasis><Emphasis Type="Italic">vulgare</Emphasis> L.) grain optimized using response surface methodology in hot air roasting

Response surface methodology was used to predict optimum conditions for hot air roasting of barley grains (temperature, time, and amount). Antioxidant capacity in the grains was highest under optimum conditions of 250 °C, 63.5 min and 42 g (one and a half layers). A correlation of R ² = 0.74 (p < 0.05) was found between 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity and total phenolic contents. Ethanol and aqueous extracts were prepared from grains roasted under optimum conditions and assessed for antioxidant capacity. Antioxidative compounds in the extracts were then identified using GC–MS. The IC₅₀ value of ethanol extract was significantly lower (11.45 μg mL^–1) than that of aqueous extract (33.54 μg mL^–1) and α-tocopherol (12.6 μg mL^–1) but higher than BHT (9.59 μg mL^–1). The same trend was observed in linoleic acid assay. In reducing power, the ethanol extract and α-tocopherol were not significantly different. Phenolic acids p-hydroxybenzaldehyde, vallinic and gallic acids were identified as the major compounds in the extracts. The results obtained from this study show that it is possible to optimize antioxidant capacity in barley grains during roasting.     

Immobilization and characterization of naringinase for the hydrolysis of naringin

The degree of hydrolysis of naringin was investigated at various temperatures (40, 50, 60 °C), enzyme concentrations (0.01–0.30 mg ml⁻¹), and pH values (2.5–5.5) for naringinase enzyme. Naringinase was immobilized on celite by simple adsorption. Naringin content was determined by HPLC method. The degree of hydrolysis of naringin showed a linear increase up to an enzyme concentration of 0.2 mg ml⁻¹ that corresponds to 82% hydrolysis. The optimum values of pH for the hydrolysis of naringin were 4.0 for free and 3.5 for immobilized enzymes. Maximum enzyme activities were found to be 70 and 60 °C for free and immobilized enzymes, respectively. The values of K _m,app and V _max,app calculated were 1.22 mM and 0.45 μmol min⁻¹ mg enzyme⁻¹ for free and 2.16 mM and 0.3 μmol min⁻¹ mg enzyme⁻¹ for immobilized enzyme, respectively. The mathematical modelling was applied to the experimental data for hydrolysis of naringin as a function of time at 30, 40 and 50 °C. The increase in temperature from 30 to 50 °C increased the rate constant 3.09 times for free enzyme. However, the rate constants found for immobilized enzyme applications did not increase in a similar trend as a function of temperature. The retained activity of celite-adsorbed naringinase was found to be 83% at their optimum conditions. The retained activity of immobilized enzyme was followed up to the fifth run and was found to be almost unchanged after the third use at optimum reaction conditions (pH 3.5, 60 °C).     

Production and Application of Xylanase from Penicillium sp. Utilizing Coffee By-products

The lignocellulosic coffee by-products such as coffee pulp, coffee cherry husk, silver skin, and spent coffee were evaluated for their efficacy as a sole carbon sources for the production of xylanase in solid-state fermentation using Penicillium sp. CFR 303. Among the residues, coffee cherry husk was observed to produce maximum xylanase activity of 9,475 U/g. The process parameters such as moisture (50%), pH (5.0), temperature (30 °C), particle size (1.5 mm), inoculum size (20%), fermentation time (5 days), carbon source (xylose), and nitrogen source (peptone) were optimized and the enzyme activity was in the range of 19,560–20,388 U/g. The enzyme production was further improved to 23,494 U/g with steam as a pre-treatment. The extracellular xylanase from the fungal source was purified to homogeneity from culture supernatant by ammonium sulfate fractionation, DE32-cellulose with a recovery yield of 25.5%. It appeared as a single band on SDS-PAGE gel with a molecular mass of approximately 27 kDa. It had optimum parameters of 50 °C temperature, pH 5.0, K _m 5.6 mg/mL, and V _max 925 μmol mg⁻¹ min⁻¹ with brichwood xylan as a substrate. The crude enzyme hydrolysed lignocellulosic substrate as well as industrial pulp. Production of xylanase utilizing coffee by-products constitutes a renewable resource and is reported for the first time.     

Purification and characterization of hydroperoxide lyase from amaranth tricolor (Amaranthus mangostanus L.) leaves

Hydroperoxide lyase (HPL) was extracted from amaranth tricolor leaves using Triton X-100, and purified to electrophoretic homogeneity by ammonium sulfate precipitation, ion-exchange chromatography, hydrophobic interaction chromatography and hydroxyapatite chromatography. The purified HPL preparation consisted of a single band and spot with a molecular mass of about 55 kDa as shown in SDS–PAGE and 2-D PAGE, respectively; the isoelectric point was found to be about 5.4. The maximum activity of the enzyme was observed at pH 6.0 and 25 °C, respectively. The HPL showed higher activity against 13-hydroperoxy-linolenic acid compared to 13-hydroperoxy-linoleic acid. K _m value for 13-hydroperoxy-linolenic acid was 62.7 μM, and the corresponding V _max was 178.5 μM min⁻¹. The activity of HPL was significantly inhibited by nordihydroguaiaretic acid, HgCl₂ and 2(E)-hexenal but not by EDTA and hexanal.     

Comparison Between Systems for Synthesis of Fructooligosaccharides from Sucrose Using Free Inulinase from Kluyveromyces marxianus NRRL Y-7571

This work is focused on the synthesis of the fructooligosaccharides (FOS) from sucrose using free inulinase from Kluyveromyces marxianus NRRL Y-7571 in aqueous and aqueous–organic systems. The most significant variables for the aqueous–organic system were identified using a fractional factorial design. The evaluated variables were the temperature, pH, sucrose concentration, inulinase activity, aqueous/organic ratio, and the polyethylene glycol concentration. The use of sequential experimental design methodology was shown to be very useful in the optimization of the FOS synthesis by inulinase either in aqueous or aqueous–organic systems. For the aqueous–organic system, the maximum Y _FOS reached was 16.7 ± 1.1 wt.% with the following operational conditions: temperature of 40 °C, enzyme activity of 4 U mL⁻¹, organic solvent/total system ratio of 25/100, pH of 6.0, and sucrose concentration of 55%. In the aqueous system, the maximum conversion obtained was 12.8 ± 1.0 wt.% under the following conditions: 40 °C, pH 5.0, 55% sucrose, and inulinase activity 4 U mL⁻¹.     

Effect of dynamic high-pressure microfluidization at different temperatures on the antigenic response of bovine β-lactoglobulin

The antigenic response of β-lactoglobulin (β-Lg), treated by dynamic high-pressure microfluidization (DHPM) at different temperatures, was determined by an indirect competitive enzyme-linked immunosorbent assay using polyclonal antibodies from rabbit serum. DHPM treatment causes changes in the protein structure and may influence the antigenicity of β-Lg. DHPM treatment of β-Lg at 90 °C showed significant effects with the antigenic response of 5.2 μg mL⁻¹ (untreated), 45 μg mL⁻¹ (40 MPa), 79 μg mL⁻¹ (80 MPa), 132 μg mL⁻¹ (120 MPa), and 158 μg mL⁻¹ (160 MPa). In combination with temperature treatment (70–90 °C), the antigenic response enhanced as the temperature increased at 160 MPa. The β-Lg antigenicities were about 14, 108, and 158 μg mL⁻¹ at 70, 80, and 90 °C, respectively. However, the influence of DHPM pressures on the antigenic response of β-Lg standards was different. DHPM modified β-Lg standards showed a remarkable increase in antigenicity when treated to 80 MPa. Above 80 MPa, the antigenic response decreased.     

Characterization of purified xylanase from finger millet (<Emphasis Type="Italic">Eleusine coracana-</Emphasis>Indaf 15) malt

Xylanase (E.C. 3.2.1.8) was purified to apparent homogeneity from 96 h finger millet (Eleusine coracana, Indaf-15) malt by a three step purification procedure via ammonium sulphate fractionation, DEAE-cellulose ion exchange and Sephadex G-75 gel permeation chromatographies with a recovery of 4.0% and fold purification of 60. Xylanase, having a molecular weight of 29 ± 2 kDa was found to be monomeric on SDS-PAGE. pH optimum of the enzyme was found to be in the range of 5.0–5.5. The activation energy was 25 kJmol⁻¹. Xylanase showed maximum stability at 35 °C in a pH range of 5.0–6.0. K _m and V _max of purified xylanase were found to be 0.2% and 4.5 μmol min⁻¹, respectively. Metal ions such as Ca²⁺, Mg²⁺, Mn²⁺, Cu²⁺, Fe²⁺, Ag²⁺ and Ni²⁺ enhanced xylanase activity at 5 mM concentration. p-chloromercuribenzoate, citric, oxalic and boric acids inhibited the enzyme in concentration dependent manner. The mode of action of xylanase was found to be “endo” as determined by the analysis of products liberated from larchwood xylan by ESI-MS and H¹NMR. In vitro studies using Bifidobacterium and Lactobacillus sp. confirmed the prebiotic activity of the xylo-oligosaccharides.     

Activities and sequences of the angiotensin I‐converting enzyme (ACE) inhibitory peptides obtained from the digested lentil (Lens culinaris) globulins

The aim of this study was the identification of potentially bioaccessible ACE‐inhibitory peptides obtained by in vitro gastrointestinal digestion of lentil globulins. ACE‐inhibitory peptides were purified by ion exchange chromatography and gel filtration. After the first step of purification, three peptide fractions with potential antihypertensive properties were obtained and the highest inhibitory activity was determined for the fraction 5 (IC₅₀ = 0.02 mg mL^?1). This fraction was separated on Sephadex G10, and six peptide fractions were obtained. The peptides of fraction (5‐F) with the highest potential antihypertensive activity (IC₅₀ = 0.13 mg mL^?1) were identified using ESI‐MS/MS. The sequences of peptides were KLRT, TLHGMV and VNRLM. Based on Lineweaver–Burk plots for the fraction 5‐F, the kinetic parameters as K_m (1.24 mm ), V_max (0.012 U min^?1), K_i (0.12 mg mL^?1) and mode of inhibition were determined.     

Determination of Melamine in Liquid Milk and Milk Powder by Titania-Based Ligand-Exchange Hydrophilic Interaction Liquid Chromatography

A simple and rapid high-performance liquid chromatography (HPLC) procedure for the analysis of melamine in liquid milk and milk powder has been developed. Decrease of acetonitrile percentage and phosphate buffer concentration in mobile phase, and lowering of buffer pH and column temperature would benefit the retention of melamine on titania. Taking advantage of the ligand-exchange and hydrophilic interaction mixed retention mode on bare titania column, neither complex pretreatment nor ion-pair reagent was required. The whole analysis for one sample including sample pretreatment and HPLC analysis could be accomplished within 30 min. The method presented good linearity (R ² = 0.9998) in a wide range of 0.02–10 μg mL⁻¹. The limit of detection (3σ) and limit of quantification (10σ) of the method were 6 and 20 μg L⁻¹, respectively, which were equivalent to 15 and 50 μg kg⁻¹ melamine in liquid milk, 60 and 200 μg kg⁻¹ melamine in milk powder, respectively. Such sensitivity could be compared with those obtained by HPLC with solid-phase extraction or HPLC coupled with tandem mass spectrometry and was adequate for the screening of melamine in tainted dairy products. The repeatability (RSDs) of the retention times and peak areas of 11 replicate detections of 1.0 μg mL⁻¹ melamine were 0.32% and 2.5%, respectively. The intermediate precision on three consecutive days (RSDs, n = 6) of the retention times and peak areas were 1.1% and 2.3%, respectively. The recovery of spiked melamine in dairy samples ranged from 95.2% to 105%. The simplicity, sensitivity and rapidity of the proposed method make it an effective alternative detecting technique for melamine.     

Aqueous medium enzymatic preparation of <Emphasis Type="SmallCaps">l</Emphasis>-alpha glycerylphosphorylcholine optimized by response surface methodology

The ability of Rhizopus chinensis lipase (Thermo 4S-3) to catalyze the deacylation of l,2-diacyl-sn-glycero-3-phosphocholines (sn-1,2-PC) for l-alpha glycerylphosphorylcholine (l-α-GPC) preparation was investigated. Response surface methodology (RSM), based on a modified central composite rotatable design, was employed to examine the effects of substrate concentrations, temperature, enzyme loading, and dosage of CaCl₂ on the l-α-GPC yield. RSM analysis indicated good correlation between experimental and predicated values. The optimal condition was confirmed as follows: reaction time, 3.5 h; temperature, 43 °C; enzyme loading, 28.2 U mL⁻¹; substrate concentration, 51.5 mg mL⁻¹; and dosage of CaCl₂, 1.9 mg mL⁻¹. Under these conditions, the l-α-GPC yield increased by 96.8%, which was close to the amount predicted by the model.     

Electrochemical sensor based on Prussian blue nanorods and gold nanochains for the determination of H<Subscript>2</Subscript>O<Subscript>2</Subscript>

In this paper, a novel electrochemical sensor for the detection of H₂O₂ was proposed based on gold nanochains and prussian blue nanorods (PB@MWCNTs), which were synthesized with multiwall carbon nanotubes (MWCNTs) as a template. With the introduction of MWCNTs and the gold nanochains, the proposed system shows synergy among the Prussian blue (PB), MWCNTs, and the gold nanochains, which amplifies the H₂O₂ sensitivity greatly. A linear range from 1.75 × 10⁻⁶ to 1.14 × 10⁻³ M with a detection limit of 0.5 × 10⁻⁶ M (S/N = 3) and a high sensitivity 300 μA mM⁻¹ cm⁻² for H₂O₂ detection is obtained. Moreover, the sensor exhibits good repeatability and stability.     

Polycyclic aromatic hydrocarbons (PAHs) in traditional and industrial smoked beef and pork ham from Serbia

Smoked beef and pork ham samples were analysed during process of smoking (after packing and storing) for the presence of the 16 EU priority PAHs via Fast GC/HRMS method. This study showed that there are differences in PAH contents between final smoked beef ham samples from traditional smokehouse (TS) (3.9 μg kg⁻¹) and industrial smokehouse (IS), (1.9 μg kg⁻¹). Also there is a difference in PAH contents in final smoked pork ham samples (4.9 μg kg⁻¹, TS; 4.2 μg kg⁻¹, IS). In beef and pork ham samples from the same smokehouse different PAH contents were observed during smoking. The highest content of examined PAHs in all beef and pork ham samples during smoking showed benzo[c]fluorene (BcL) (beef ham: from 0.3 μg kg⁻¹ to 1.5 μg kg⁻¹; pork ham: from 0.2 μg kg⁻¹ to 2.1 μg kg⁻¹).The maximum level for benzo[a]pyrene (BaP) of 5 μg kg⁻¹ in smoked meat products was not exceeded in any samples. Correlation statistic analysis (P < 0.05) of obtained contents from samples both from TS and IS showed that BaP is a good marker both for 16 EU priority PAHs and 12 IARC probably and possibly carcinogenic PAHs (IS: R _BaP/Σ16PAHs = 0.95, R _BaP/Σ12PAHs = 0.96; TS: R _BaP/Σ16PAHs = 0.71, R _BaP/Σ12PAHs = 0.88).     

A Sensitive, Selective New Analytical Reagent, 2, 6-Diacetylpyridine bis-4-phenyl-3-thiosemicarbazone for Extractive Spectrophotometric Determination of Mo(VI) in Food Samples

Synthesized a new reagent 2, 6-Diacetylpyridine bis-4-phenyl-3-thiosemicarbazone (2, 6-DAPBPTSC) characterized and is proposed as a sensitive and selective analytical reagent for the extractive spectrophotometric determination of molybdenum(VI) at pH 3.5 to form a yellowish orange colored 1:1 chelate complex. The absorbance was measured at a maximum wavelength, 500 nm. This method obeys Beer’s law in the concentration range 0.90–9.00 μg mL⁻¹ and the correlation coefficient of Mo(VI)–2,6-DAPBPTSC complex is 0.954, which indicates an adequate linearity between the two variables with good molar absorptivity and Sandell’s sensitivity, 1.212 × 10⁴ L mol⁻¹cm⁻¹, 0.0079 μg cm⁻², respectively. The precision and accuracy of the method is checked with calculation of relative standard deviation (n = 5), 0.894% and the detection limit value is 0.0056 μg mL⁻¹. The instability constant of the method has been calculated by Asmus’ method as 6.476 × 10^–5, at room temperature. The interfering effect of various cations and anions has also been studied. The method was successfully applied for the determination of Mo(VI) in food and water samples. The validity of the present method was evaluated in terms of Student ‘t’ test and Variance ‘f’ test, which indicates the significance of the present method an inter comparison of the experimental values, using atomic absorption spectrometer.     

Determination of reaction kinetics of hydrolysis of tilapia (Oreochromis niloticus) protein for manipulating production of bioactive peptides with antioxidant activity,angiotensin‐I‐converting enzyme inhibitory activity and Ca‐binding properties

To manipulate enzymatic hydrolysis of tilapia (Oreochromis niloticus) muscle protein for production of bioactive peptides, its reaction kinetics was intensively studied. The study showed that the production of peptides with different bioactive properties including antioxidant activity, angiotensin‐I‐converting enzyme (ACE) inhibition and Ca‐binding property and their kinetics were affected by the degree of hydrolysis and substrate concentration. A comparative study on reaction kinetics found that the kinetic parameters for the production of each bioactive peptide are unique, that is, the maximum initial velocity, V_max, for hydrolysis of protein was as high as 1.07 mg mL^?1 min^?1, but that for the production of peptides with antioxidant activity and Ca‐binding property were very low, range of 7.14–66.7 μg mL^?1 min^?1, and that for the production of peptides with ACE inhibitory activity was the lowest, at 2.57 μg mL^?1 min^?1. This knowledge of reaction kinetics of protein hydrolysis would be useful for manipulating and optimising the production of peptides with desired bioactive properties.