SEED LIPIDS OF THE MIRACLE FRUIT (SYNSEPALUM DULCIFICUM)

The lipids from the Miracle Fruit seeds comprised 10.15% of the dry weight. The neutral lipids, phospholipids and glycolipids were separated by silicic acid chromatography and the fatty acid composition of each fraction determined. The non-saponifiable lipids, amounting to 1.6% of the neutral lipids were separated into several fractions by thin-layer chromatography and the components of each analyzed by combined gas chromatography-mass spectrometry. The hydrocarbon fraction contained the n-alkane series from C₁₇ to C₃₂ with the C₂₉ and C₃₁ members present in relatively large quantities.
The major triterpene alcohols were α-and β-amyrin and the major sterol was identified as δ⁷ spinasterol. Two short-chain alchols were also present.     

Quenching Mechanisms and Kinetics of α-Tocopherol and β-Carotene on the Photosensitizing Effect of Synthetic Food Colorant FD&C Red No. 3

ABSTRACT: Effects of FD&C Red No. 40, Red No. 3, Yellow No. 5, Yellow No. 6, Green No. 3, Blue No. 1 and Blue No. 2 on 0.03M soybean oil oxidation in acetone at 25 °C under light were studied by measuring headspace oxygen depletion. As Red No. 3 increased from 0 to 5, 20, 100 and 200 ppm, the headspace oxygen was reduced by 2 to 70, 73, 77 and 77%, respectively, for 4 h. Only Red No. 3 acted as a photosensitizer to produce singlet oxygen in the oil. The quenching rates of α-tocopherol and β-carotene for the singlet oxygen by Red No.3 were 4.1 × 10⁷ M⁻¹s⁻¹ and 7.3 × 10⁹ M⁻¹s⁻¹, respectively. When β-carotene was below 1.86 × 10⁻⁶ M, β-carotene quenched singlet oxygen, but it quenched both singlet oxygen and Red No. 3 at or above 3.72 × 10⁻⁶ M. However, α-tocopherol quenched singlet oxygen only.     

ISOLATION OF β2-CHACONINE, A POTATO BITTERNESS FACTOR

The 4-rhamnoglucoside of alkaloid solanidine, β₂-chaconine, was isolated from a mixture of α-solanine with β₂-chaconine by fractionation from hot ethyl acetate. The mixture of the two glycoalkaloids is obtained by endogenous enzyme action in a homogenate of potato berries and blossoms within a 24–36 h incubation at 37° C, pH 6.00. The identity of β₂-chaconine was ascertained by m.p., [α]D° IR spectrum, TLC of sugar components and reference to partial acid hydrolysis products of α-chaconine.     

THE MOLECULAR WEIGHT OF α-AMYLASE INHIBITOR FROM WHITE BEAN cv 858B (PHASEOLUS VULGARIS L.) IS 56 kDa, NOT 20 kDa

The molecular weights (M_rs) of α-amylase inhibitors (αAIs)fiom 18 (Ah) bean cultivars estimated by Superose 12 gelfiltration chromatography were 22–62% smaller than those determined by Sephadex G-100 gel filtration and by polyacrylamide gel electrophoresis (PAGE) methods. αAI-4 from WKB cultivar 858B was purified and the Mr was shown to be 51.0 kDa based on Sephadex G-100 gel filtration chromatography and by PAGE. A M_r for aAI-4 of 56.714 kDa was determined by laser-assisted time-of-flight mass spectrometry and appears to be the true M_r of the mature glycosylated active aAI-4. The results show that Superose 12 gelfiltration chromatography is not usefulfor M_r determination of some proteins. Sodium dodecyl sulfate electrophoresis (SDS-PAGE) showed that the 56.7 kDa aAI-4 molecule dissociated into 45.0, 33.6,15.2 and 12.4 kDa submolecules, with only the two small subunits, a and β, present at high SDS concentration. This provides evidence that the aAI-4 molecules composition is α₂β².     

RHEOLOGICAL AND CHEMICAL PROPERTIES OF MOZZARELLA CHEESE

Dynamic viscoelastic parameters and chemical properties of Mozzarella cheese produced using a "no-brine" cheese making method with 3 different cooking temperatures (38, 41, and 44C) were determined. Samples were stored for 3 weeks at 4C before dynamic mechanical analysis at 22C. G', G" and tan δ were 5.8 – 6.4 × 10⁵ dyne/cm², 1.9 – 2.1 × 10⁵ dyne/cm², and 0.33 – 0.35, respectively, at 1% strain and 10 rad/s. The percentage of intact α_s-casein and β-casein were 38–40% and 33–35% of total protein in the cheese, respectively. The range of cooking temperatures used in this experiment had little effect on dynamic viscoelastic properties or the amount of intact protein for the cheese.     

EXAMINATION OF THE MICROBIAL STATUS OF SELECTED INDIGENOUS FERMENTED FOODS IN NIGERIA

Four Nigerian traditionally fermented foods (wara, nono, ogi and kununzaki) were evaluated for the presence of some microorganisms of public health concern. Among the dairy foods , Staphylococcus aureus and Klebsiella sp. were isolated from wara while Escherichia coli, Salmonella sp. and Klebsiella sp. were isolated from nono. The cereal-based fermented foods (ogi and kunu-zaki) contained Bacillus subtilis, E. coli, S. aureus, Klebsiella sp. and Enterococcus faecalis. The mesophilic aerobic counts were: 5 × 10⁵ for wara; nono, 1.53 × 10⁷; ogi, 3.6 × 10⁶ and kunu-zaki, 2.6 × 10⁶ cfu/mL. The enterobacteriaceae counts on nono, wara, ogi and kunu-zaki were 1.79 × 10⁷, 4.5 × 10⁵, 4.0 × 10⁵ and 1.2 × 10⁶ cfu/mL, respectively. No Vibrio count (detection limit: <10 cfu/mL) was recorded in all the food samples considered. The yeast and mold counts ranged from 1.0 × 10⁵– 3.31 × 10⁷ among the food products. The antimicrobial susceptibility patterns of the organisms isolated from dairy products (nono and wara) revealed that they were resistant to ampicillin (100%) and sensitive to gentamicin (100%) and nalidixic acid (100%). Most isolates from cereal based products (ogi and kunu-zaki) were 100% resistant to penicillin, ampicillin and chloramphenicol. This work highlights the need to maintain hygienic standards in the preparation of our locally fermented cereal and dairy foods.     

EFFECT OF SEVERAL EXPERIMENTAL PARAMETERS ON COMBINATION OF RED KIDNEY BEAN (PHASEOLUS VULGARIS) α-AMYLASE INHIBITOR WITH PORCINE PANCREATIC α-AMYLASE

Purified red kidney bean (Phaseolus vulgaris) amylase inhibitor forms a 1:1 stoichiometric complex with porcine pancreatic α-amylase leading to complete loss of enzyme activity on starch. Rate of complex formation is pH dependent and is maximal at pH 5. The rate constants for complex formation, as measured by loss of amylase activity, were 2.85 × 10⁴ M^-1 sec^-1 at pH 6.9 (ionic strength of 0.918) and 2.55 × 10⁵ M^-1 sec^-1 at pH 5 at 30°C. At pH 6.9, rate of complex formation was 4.8 times faster at 0.918 ionic strength as compared with the rate at 0.138 ionic strength. At 30°C, pH 6.9 and ionic strength of 0.168 the dissociation constant of the enzyme-inhibitor complex was determined to be 3.5 × 10^-11 M. The rate constant for dissociation of the complex was calculated to be 8.7 × 10^-8 sec^-1 under the same conditions. The rate constant for complex formation, at ionic strength of 0.168, was 1.1 × 10⁴ M^-1 sec^-1 at 37⁰ and 9.77 × 10² M^-1 sec^-1 at 25.7°C. The calculated activation energy for complex formation is 39.5 kcal/mole suggesting a rate-controlling conformational change. Oxidation of the carbohydrate moiety of the glycoprotein inhibitor caused complete loss of activity. Maltose, a competitive inhibitor of α-amylase, bound as readily to the enzyme-inhibitor complex as to free α-amylase. Trypsinized α-amylase, although still able to bind to Sephadex, did not bind inhibitor. The experiments with maltose and trypsinized amylase suggest the inhibitor may not bind at the active site of α-amylase.     

AN ESTEROLYTIC ACTIVITY FROM A WILD EDIBLE MUSHROOM, LYCOPERDON PERLATUM

Lycoperdon perlatum Pers. (Lycoperdaceae, Agaricales, Agaricomycetidae, Agaricomycetes, Basidiomycota, Fungi) was evaluated for its esterolytic potential. Native electrophoresis of the crude extracts showed four bands having Rf values of 0.34, 0.39, 0.52 and 0.59. The esterase showed the highest activity toward a short-chain substrate, p -nitrophenyl acetate. Optimum reaction conditions for L. perlatum crude extract were attained at pH 8.0 and 40C. Esterolytic activity of enzyme extract was stimulated in the presence of Mn^{2 +} , Fe^{2 +} , Ca^{2 +} and Zn^{2 +} in the reaction mixture. The enzyme activity was stimulated by incubation at pH 6.0 but retained 77% of its original activity at its optimum pH after 24 h. Thermal inactivation was displayed after incubation for 20 min at various temperatures above 30C. At 1 mM final concentration, 2-mercaptoethanol, dithiothreitol, ethylenediamine tetraacetic acid and p -methylphenyl sulfonylfluoride inhibited the esterolytic reaction. These results support that the crude L. perlatum extract possesses an esterolytic activity having properties similar to other esterases.

PRACTICAL APPLICATIONS

Esterases catalyzing the cleavage and formation of ester bonds are known α/β-hydrolases (EC 3.1.1.X). Esterases are used for the synthesis of flavor esters for the food industry, modification of triglycerides for fat and oil industry and resolution of racemic mixtures used for the synthesis of fine chemicals for the pharmaceutical industry. Therefore, the search for new enzyme sources is important for the development of new enzymes and applications.     

Effect of technological parameters on the characteristics of kasseri cheese made from raw or pasteurized ewes' milk

Two groups of kasseri cheese (pasta filata type) were manufactured from raw or pasteurized ewes' milk, without starter cultures. Cheeses of each group were divided into two subgroups: the first was ripened and stored at 4°C and packaged in plastic film; the second ripened and stored at 15°C and coated with paraffin wax. Milk pasteurization and technological parameters had a significant effect on the pH ( P  < 0.05), while only technological parameters had an effect on the total solids content. At day 120, the range of mean cfu/g counts for the mesophilic aerobic flora was 9.5 × 10⁷−1.4 × 10⁸; for the thermophilic streptococci, the range was 2.6 × 10⁷−7.6 × 10⁷; and for the thermophilic bacilli, 9.8 × 10⁶−1.7 × 10⁷. Changes in the N fractions became significant after 30 days of ripening. For mature 120-day-old cheeses, the percentage of total N soluble at pH 4.6 was 22.7%–22.9% in raw milk cheeses and 19.0%–21.7% in pasteurized milk cheeses. The percentage of total N soluble at 12% TCA was 10.1%–12.2% in raw milk cheeses and 7.3%–11.5% in pasteurized milk cheeses; the percentages of total N soluble at 5% PTA were 3.1%–4.0% and 2.6%–3.6%, respectively. The residual α_s-casein percentages at day 120 ranged between 63% and 78% of the respective area at day 1; the residual β-casein ranged between 67% and 75%. There were some characteristic differences in the reverse phase-HPLC peptide profiles of the four cheeses. In general, the effect of the different ripening conditions was more pronounced in cheeses made from pasteurized milk.     

SURFACE PROPERTIES OF PROTEIN LAYERS ADSORBED FROM MIXTURES OF GELATIN WITH VARIOUS CASEINS

We report surface tensions and surface shear viscosities of protein layers adsorbed from mixtures of gelatin +α_s11-casein, β-casein, k-casein or sodium caseinate. Under conditions where the two protein components carry opposite net charges, the combined experimental data at 25C and neutral pH are consistent with a two-layer model of the mixed protein film. The surface properties of sodium caseinate lie intermediate between those for the two major individual caseins (α_s1 and β).     

STRUCTURAL FEATURES OF RED KIDNEY BEAN á-AMYLASE INHIBITOR IMPORTANT IN BINDING WITH á-AMYLASE

The structural features of the glycoprotein α-amylase inhibitor from red kidney bean (Phaseolus vulgaris) important for binding to α-amylase were examined. The inhibitor contained 13.0% carbohydrate. The carbohydrate portion of the inhibitor is composed of 25 mannose, 2 xylose, 1 fucose and 17 N-cetylglucosamine residues per mol. Seventy (70) percent of the neutral carbohydrates were removed from the inhibitor by treatment with endo-β- N -acetylglucosaminidase H. The carbohydrate remaining attached to the protein consisted of 7 mannose, 2 xylose, and 1 fucose residues (N-acetylogucosamine was not determined). The intact glyco chains obtained from the pronase digest of the inhibitor failed to inhibit β-amylase (3.3 times 10⁵ fold molar excess of glyco chains to enzyme) when preincubated with the enzyme at 30°C for 30 min at pH 6.9 before adding starch as substrate.
Oxidation of one tryptophan residue of the α-amylase inhibitor with N-bromosuccinimide at pH 6.0 led to a 50% loss in inhibitory activity. Periodate oxidation of α-amylase inhibitor led to less of two tyrosine and one methionine residues per mole of inhibitor within 15 min. There was not a direct correlation between modification of these residues and loss of inhibitory activity. Modification of three of the five histidine residues with diethylpyrocarbonate resulted in about 50% loss in inhibitory activity.     

Singlet Oxygen Oxidation Rates of ∝-, γ-, and δ-Tocopherols

ABSTRACT:  The reaction rate constants of 5 × 10⁻⁴ M, 10 × 10⁻⁴ M, and 20 × 10⁻⁴ M α-, γ-, and δ-tocopherols with singlet oxygen in methylene chloride containing 1 × 10⁻⁵ M chlorophyll under light at 25 °C for 60 min were studied. The oxidation of tocopherols determined by a spectrophotometric method showed that the losses of 20 × 10⁻⁴ M α-, γ-, and δ-tocopherols after 60 min under light were 21%, 16%, and 9%, respectively. The degradation of α-, γ-, and δ-tocopherols was undetectable in the absence of chlorophyll under light or in the presence of chlorophyll in dark. The losses of tocopherols under light were mainly due to singlet oxygen oxidation. The degradation rates of 20 × 10⁻⁴ M α-, γ-, and δ-tocopherols were 6.6 ×10⁻⁶ M/min, 5.0 × 10⁻⁶ M/min, and 2.9 × 10⁻⁶ M/min, respectively. The reaction rates between α-, γ-, or δ-tocopherol and singlet oxygen were 4.1 ×10⁶/M s, 3.3 × 10⁶/M s, and 1.4 × 10⁶/M s, respectively. The singlet oxygen oxidation rate of δ-tocopherol was significantly lower than α- or γ-tocopherol at α= 0.05. As the electron density in the chromanol ring of tocopherol increased, the singlet oxygen oxidation was increased.     

Comparative evaluation of the olive oil given by a new processing system

To evaluate the level of quality of the olive oil given by a new processing system we compared it with the percolation oil, which is known to be of high quality. The new product was characterized by the following: (i) higher contents of tocopherols, tyrosol- and hydroxytyrosol-aglycones, volatile aromatic substances, sterols, triterpene alcohols and lipochromes, and higher values of 1,2-diglycerides/1,3-diglycerides and campesterol/stigmasterol ratios, colour indices and turbidity; (ii) lower contents of waxes and triterpene dialcohols, and lower values for spectrophotometric indices, carbonyl index, Wolff's ratio (K₂₃₂/K₂₇₀), acidity, and peroxide number; (iii) similar contents of phenols, o- diphenols, free tyrosol, free hydroxytyrosol, triglycerides, stigmastadienes, and aliphatic alcohols, similar sensory score, and similar value of resistance to autoxidation, alcohol index, overall quality indices, and fatty acid composition.     

MICROBIOLOGICAL QUALITY OF TRADITIONAL MARKET CURED FISH IN TANZANIA

The microbiological quality of six varieties of retail market traditionally cured fish in Morogoro, Tanzania was investigated over a five-month period. The fish were contaminated with bacteria and molds at levels of: total aerobes, 10⁶ - 1.7 × 10⁷ c.f.u/g; faecal coliforms, 1.1 × 10¹ - 2.5 × 10³ MPN/g; faecal streptococci, 1.4 × 10¹ - 1.3 × 10³ MPN/g; Staphylococcus aureus, 1.3 × 10³ - 8.6 × 10³ c.f.u/g; Aspergillus flavus group, 2.1 × 10¹ - 2 × 10² c.f.u/g of fish. Of faecal coliform, 45% of the isolates were Escherichia coli. Twenty-five percent of the S. aureus isolates were coagulase positive. Sixteen percent of A. flavus isolates were aflatoxigenic. Aflatoxin contamination ranged from O to 18.5 μg/kg of fish. Insect infestation by Dermestes spp. and mites was observed. The results of this preliminary study emphasize the importance of proper processing and handling offish in the tropics in order to safeguard public health .     

PREPARATION OF ENRICHED FRACTIONS OF α-LACTALBUMIN AND β-LACTOGLOBULIN FROM CHEESE WHEY USING CARBON DIOXIDE

A new process for preparing enriched fractions of α-lactalbumin (α-La) and β-lactoglobulin (β-Lg) from whey protein concentrate was developed. To prepare the fractions, solutions containing from 7% (w/w) to 25% (w/w) whey protein concentrate (WPC75) were sparged with carbon dioxide (CO₂) in a batch reactor. The effects of pressure, temperature, concentration, and residence time on the distribution of the individual whey proteins in each fraction were investigated. Best results were obtained for 7% (w/w) WPC75 solutions, at reactor fill pressures of 4140 kPa and 5520 kPa, temperature of 64C and reactor residence time of 10 min. Gel electrophoresis of unpurified enriched samples showed that recovery of α-La was approximately 55% and that of β-Lg was 78%. The resulting fractions have a pH of 6.0 and contain no added salts.     

The yeast flora of stored ready-to-use carrots and their role in spoilage

Spoilage of ready-to-use grated raw carrots packaged in polymeric films and stored at 10°C was investigated for involvement of yeasts. Cryptococcus albidus was only isolated during the first 3 days of storage, increasing to levels of 10⁵–10⁶g^-1. Candida lambica was more commonly isolated after 3–7 days of storage, and reached 10⁷–10⁸g^-1 after 12 days. Candida sake was present throughout storage, increasing from 10⁵–10⁶ after 3 days to 10⁷–10⁸ after 12 days. In some samples, Candida parapsilosis and Candida tropicalis were also isolated at levels similars to C. sake . All the yeasts isolated at the end of storage were fermentative species and their metabolism was characterized with a Warburg apparatus. Neither the number of yeasts nor the composition of the yeast flora were related to the deterioration of the product. Although Candida lambica inoculated on grated carrots caused spoilage after 12 days at 10°C, the high O₂ permeable film was most effective in reducing exudate.     

EFFECT OF ADDITION OF α-, β-, AND κ-CASEIN, AND Na-CASEINATE ON VISCOELASTIC PROPERTIES OF SKIM MILK CURD

Creep compliance was used to determine the effects of the addition of α-, β-, and κ-casein, and Na-caseinate on the viscoelastic properties of skim milk curd. The results of all measurements can be represented by a six-element mechanical model. Addition of α- and β-casein, and Na-caseinate (1.80g/L) to raw skim milk reduced the instantaneous modulus of rigidity and final viscosity of the curd, while κ-casein addition at the same level increased both viscoelastic parameters. Shielding of κ-casein and depletion of serum Ca⁺⁺ ions by α- and β-casein is thought to have caused the reduction of curd rigidity and viscosity. Subsequent experiments indicated that the addition of β-casein before and after rennet hydrolysis produced different curd strength with the latter producing a stronger curd.     

Effects of Lactic Acid Bacteria and Fermented Milks on Eicosanoid Production by Intestinal Epithelial Cells

ABSTRACT: Fermented milk products produced with probiotic lactic acid bacteria (LAB) have attracted interest due to their potential health benefits. Probiotic bacteria have a range of immunomodulatory activity, interacting with a variety of cell types in the immune system. Interactions with intestinal epithelial cells (IEC) are an avenue through which probiotics and their fermented milks can influence production of key immunoregulatory molecules, including cytokines and eicosanoids. The eicosanoids, which include the prostaglandins (PGs), are lipid mediators implicated in both acute and chronic inflammatory processes. The primary objective of this study was to determine the ability of probiotic LAB and their ferments to interact with IEC and influence their eicosanoid production. Effects of LAB and their milk ferments on prostaglandin E₂ (PGE₂) and prostaglandin F_2α (PGF_2α) production by human IEC lines were determined using a competitive enzyme immunoassay. LAB alone did not alter interleukin (IL)-1β-induced prostaglandin production by IEC. However, milk fermented with Lac-tobacillus (L.) rhamnosus strain R0011 significantly suppressed IL-1β-induced levels of PGE₂ and PGF_2α, an effect which was counteracted by the addition of strain R0011. Milk ferments prepared withL. acidophilus strain R0052 were less effective in down-regulation of PG production by IEC. Naltrexone, an opioid receptor antagonist, blocked the suppressive effects of L. rhamnosus R0011 milk ferments on PGF_2α production by IEC, suggesting that the bioactivity the ferments is opioid receptor-mediated. These findings support immunomodulatory potential of fermented food components through interactions with intestinal epithelial cells.     

Production of hard cheese from caprine milk by the use of two types of probiotic cultures as adjuncts

Hard cheeses (Kefalotyri-like) were manufactured from caprine milk with yoghurt as a starter (A), and with its partial replacement with the probiotic adjuncts Lactobacillus rhamnosus LC 705 (B) and/or Lactobacillus paracasei ssp. paracasei DC 412 (C). Both adjuncts retarded the growth of enterococci, and the environment in cheese B did not favour the recovery of lactic acid bacteria (LAB) on Rogosa agar. However, better recovery of the LAB population on M17 agar from cheeses B and C made with adjuncts was recorded early in ripening, and this was accompanied by a greater decrease in pH. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of whole-cell protein demonstrated that cheese C, made with Lb. paracasei ssp. paracasei as adjunct, is a better vehicle for delivery of live probiotic cells (10 ⁷   cfu/g) to the gastrointestinal tract than cheese B, made with Lb. rhamnosus ; the latter did not belong to the predominant microflora of one out of the two B cheeses. Urea-PAGE electrophoresis results indicated that adjunct lactobacilli enhanced the degradation of both α _S -casein (α _S -CN) and β-casein (β-CN). In the fresh cheese, hydrolysis of α _S -CN was more rapid than β-CN, and the free amino acid content of B and C was higher than in A. Lipolysis products were also higher in B and C than in A as ripening progressed, and the organoleptic characteristics of these cheeses resulted in higher scores, in the order C > B > A. Thus, making Kefalotyri-like cheese from caprine milk with probiotic lactobacilli, particularly Lb. paracasei ssp. paracasei, as adjunct can be considered an effective way of producing a cheese with a large number of probiotic cells.     

Riboflavin Photosensitized Singlet Oxygen Oxidation of Vitamin D

Samples containing various levels of vitamin D and riboflavin, with and without ascorbic acid or a-tocopherol were prepared in a model system. Samples were stored in the light or in the dark at 45®C for up to 8 h. Headspace oxygen was determined by gas chromatography with thermal conductivity detection. Oxidation of vitamin D was not observed in samples without riboflavin stored in the light nor in samples with riboflavin stored in the dark. Vitamin D with riboflavin was oxidized under light. α-Tocopherol quenched singlet oxygen at a rate of 2.52 × 10⁸ M^-1 sec^-1, whereas ascorbic acid quenched singlet oxygen at a rate of 2.23 × 10⁷ M^-1 sec^-1.