Isolation and characterisation of a novel angiotensin I‐converting enzyme‐inhibitory peptide derived from douchi,a traditional Chinese fermented soybean food

BACKGROUND: Douchi, a traditional fermented soybean food, has recently attracted a great deal of attention owing to its superior physiological activity. In the present study the angiotensin I‐converting enzyme (ACE)‐inhibitory activity of typical douchi procured from various regions of China was analysed. An ACE‐inhibitory peptide derived from the most potent douchi was also isolated and characterised. The pattern of ACE inhibition and resistance to hydrolysis by gastrointestinal proteases of this peptide are described. RESULTS: ACE‐inhibitory activities were detected in all douchi samples, with IC₅₀ values ranging from 0.204 to 2.011 mg mL^?1. Among the douchi samples, a Mucor‐type douchi exhibited the most potent ACE‐inhibitory activity (IC₅₀ = 0.204 mg mL^?1). A novel ACE‐inhibitory peptide was then isolated from this Mucor‐type douchi using ultrafiltration followed by Sephadex G‐25 column chromatography and reverse phase high‐performance liquid chromatography. The amino acid sequence of the purified peptide was identified by Edman degradation as His‐Leu‐Pro (IC₅₀ = 2.37 µmol L^?1). The peptide is a competitive inhibitor and maintained its inhibitory activity even after incubation with some gastrointestinal proteases. CONCLUSION: The present study shows that peptides derived from soybean fermentation during douchi processing could be the main contributor to the ACE‐inhibitory activity observed. Copyright © 2009 Society of Chemical Industry     

Antioxidant and melanogenesis‐inhibitory activities of collagen peptide from jellyfish (Rhopilema esculentum)

BACKGROUND: Jellyfish collagen was hydrolysed with trypsin and properase E, and jellyfish collagen peptide (JCP) was purified from the enzymatic hydrolysate using ion exchange chromatography and gel filtration. The antioxidant activity of JCP in a linoleic acid emulsion system, its superoxide anion‐ and hydroxyl radical‐scavenging activities and its copper‐chelating ability were evaluated in vitro. Initial investigations of JCP's ability to inhibit melanogenesis were carried out using cultured B16 melanoma cells. RESULTS: The molecular weight distribution of JCP was from 400 to 1200 Da. Amino acid analysis showed that JCP was rich in Gly, Pro, Ser, Ala, Glu and Asp and had a total hydrophobic amino acid content of 384.2 g kg^?1. JCP showed high antioxidant activity (IC₅₀147.8 µg mL^?1), superoxide anion‐scavenging activity (IC₅₀21.9 µg mL^?1), hydroxyl radical‐scavenging activity (IC₅₀16.7 µg mL^?1) and copper‐chelating ability (IC₅₀88.7 µg mL^?1) in vitro. It also significantly inhibited intracellular tyrosinase activity, decreased melanin content and enhanced glutathione synthesis (P < 0.05). Furthermore, JCP decreased intracellular cAMP levels and suppressed tyrosinase mRNA expression. CONCLUSION: Based on the results of this study, JCP exerts anti‐melanogenic actions via its antioxidant properties and copper‐chelating ability. JCP could be used as a natural skin‐lightening agent in the medicine and food industries. Copyright © 2009 Society of Chemical Industry     

Angiotensin‐I converting enzyme inhibitory and antioxidant activities of peptide fractions extracted by ultrafiltration of cowpea Vigna unguiculata hydrolysates

BACKGROUND: Enzymatic proteolysis of food proteins is used to produce peptide fractions with the potential to act as physiological modulators. Fractionation of these proteins by ultrafiltration results in fractions rich in small peptides with the potential to act as functional food ingredients. The present study investigated the angiotensin‐I converting enzyme (ACE‐I) inhibitory and antioxidant activities for hydrolysates produced by hydrolyzing Vigna unguiculata protein extract as well as ultrafiltered peptide fractions from these hydrolysates. RESULTS: Alcalase^®, Flavourzyme^® and pepsin–pancreatin were used to produce extensively hydrolyzed V. unguiculata protein extract. Degree of hydrolysis (DH) differed between the enzymatic systems and ranged from 35.7% to 58.8%. Fractionation increased in vitro biological activities in the peptide fractions, with IC₅₀ (hydrolysate concentration in µg protein mL^?1 required to produce 50% ACE inhibition) value ranges of 24.3–123 (Alcalase hydrolysate, AH), 0.04–170.6 (Flavourzyme hydrolysate; FH) and 44.7–112 (pepsin–pancreatin hydrolysate, PPH) µg mL^?1, and TEAC (Trolox equivalent antioxidant coefficient) value ranges of 303.2–1457 (AH), 357.4–10 211 (FH) and 267.1–2830.4 (PPH) mmol L^?1 mg^?1 protein. CONCLUSION: The results indicate the possibility of obtaining bioactive peptides from V. unguiculata proteins by means of a controlled protein hydrolysis using Alcalase^®, Flavourzyme^® and pepsin–pancreatin. The V. unguiculata protein hydrolysates and their corresponding ultrafiltered peptide fractions might be utilized for physiologically functional foods with antihypertensive and antioxidant activities. Copyright © 2010 Society of Chemical Industry     

A novel chitinase isolated from Vicia faba and its antifungal activity

A novel chitinase with antifungal activity was isolated from fava bean (Vicia faba) seeds. The protein exhibited a molecular mass of 21.5 kDa in reduced condition while 25.5 kDa in oxidized condition on SDS-PAGE, indicating that there are disulfide bonds inside the molecule. Its N-terminal amino acid sequence was determined to be D-D-V-G-S-V-I-S-A-S-L-F-E-Q-L-L-K-H, showing homologous to those of chitinase and chitinase precursors from leguminous plants. The optimum pH and the optimum temperature for activity toward N-acetyl-d-glucosamine were 5.4 and 50 °C, respectively. The pI was determined to be 8.7 by isoelectric focusing electrophoresis. The chitinase was thermostable up to 58 °C in both enzymatic reaction and antifungal activity. It showed chitin-binding activity, suggesting that the catalytic domain is involved in the binding of chitinase to a certain extent. In addition, it exerted potent antifungal action toward a variety of fungal species including Pythium aphanidermatum, Fusarium solani, Physalospora piricola, Alternaria alternate, Botrytis cinerea, and Fusarium oxysporum f. sp. melonis. The present findings demonstrated a novel chitinase with disulfide bonds inside the molecule and show antifungal significance in agriculture.     

Relationships between amino acid composition and nitrogen of foxtail (Italian) millet (Setaria italica) grain of different varieties

The amino acid composition of 13 samples of foxtail millet (Setaria italica L) from six Chinese and one French varieties was determined as a function of their N content (N), which ranged from 1·82 to 3·65 g per 100 g of grain DM. The levels of amino acids in grain DM increased linearly with N with correlation coefficients close to 1 for most of them regardless of foxtail millet genotype or phenotype. Thus simply knowing N enables one to predict the amino acid composition of any foxtail millet grain sample. Amino acids in crude protein of grain (g 16 g^?1N)changed as quadratic functions of N, which decreased for glycine, cysteine, tysine, histidine and arginine, remained nearly constant for valine, threonine, tyrosine, methionine and aspartate plus asparagine, and increased for other amino acids. Foxtail millet appeared as the only cereal in which lysine is the only limiting essential amino acid. However, the lysine score was low and intermediate between that of maize and sorghum, falling from 48 to 31 % when N increased from 1·82 to 3·65 g per WO g DM. The N-to-protein conversion factor strongly increased with N and was the highest of all cereals within the N range studied. The results also showed that the composition of storage proteins accumulated in grains remained constant, with a prolamin to glutelin ratio close to three and independent of grain protein content.     

Purification of citrus limonoids and their differential inhibitory effects on human cytochrome P450 enzymes

Recent studies demonstrated that citrus limonoids and flavonoids possess numerous health promoting properties. In the present study, glucosides of limonoids and flavonoids were purified from citrus molasses and limonoid aglycones from citrus seeds. Glucosides were separated on styrene (divinylbenzene), Q‐sepharose resins with increasing concentration of sodium chloride. A pH‐dependent cold precipitation was carried out for the isolation of naringin in large quantity. Major aglycones such as limonin and nomilin were isolated from seeds by direct crystallization and minor limonoids were purified by vacuum liquid chromatography. The structures of the isolated compounds were confirmed by NMR spectra. Individual limonoids were tested for O‐dealkylase and hydroxylase activities of human cytochrome P450 (CYP) isoenzymes such as CYP1A2, CYP1B1, CYP3A4 and CYP19, using ethoxyresorufin, methoxyresorufin and dibenzylfluorescein as substrates. Partial to high inhibition of CYPs was observed in dose‐dependent assays. Significant (P < 0.001) reductions in enzyme activities were observed with purified compounds above 2 µmol. Kinetic analyses indicated that limonin glucoside inhibited CYP19 competitively (IC₅₀, 7.1 µ mol L^?1), whereas Nomilinic acid glucoside inhibited it noncompetitively (IC₅₀, 9.4 µ mol^?1). Nomilinic acid glucoside was the most potent limonoid, with an overall IC₅₀ of < 10 µ mol, for all the enzymes tested. The differential inhibition of CYPs can be ascribed to structural variations of the limonoid nucleus. Limonoid inhibition of key CYPs involved in carcinogenesis supports growing evidence that citrus limonoids act as anticancer agents. Copyright © 2007 Society of Chemical Industry     

Angiotensin I‐converting enzyme inhibitory peptides in skim milk fermented with Lactobacillus helveticus 130B4 from camel milk in Inner Mongolia,China

BACKGROUND: Angiotensin I‐converting enzyme (ACE) is a dipeptidyl carboxypeptidase associated with the regulation of blood pressure. ACE inhibition results in a lowering of blood pressure. Lactic acid bacteria are known to produce ACE inhibitors during fermentation. Fermented camel milk is the main traditionally fermented dairy food for desert nomads. The beneficial effects of fermented camel milk, which include the prevention of such diseases and conditions as gastroenteritis, tuberculosis and hypertension, have been demonstrated experimentally. RESULTS: ACE inhibitory activity was observed in fermented milk containing Lactobacillus helveticus 130B4, a strain isolated from traditionally fermented camel milk. The peptide that inhibited ACE was purified from the fermented milk by reverse‐phase high‐performance liquid chromatography. The amino acid sequence of the peptide was identified as Ala‐Ile‐Pro‐Pro‐Lys‐Lys‐Asn‐Gln‐Asp (IC₅₀ = 19.9 µmol L^?1). The same Ala‐Ile‐Pro‐Pro‐Lys‐Lys‐Asn‐Gln‐Asp sequence was found in κ‐casein (κ‐CN) f107–115 from milk. The inhibitory activity of this nonapeptide (κ‐CN f107–115) was almost preserved even after successive digestion with pepsin, trypsin and chymotrypsin. Furthermore, the inhibitory activity of the purified peptide was completely preserved after heat treatment at 100 °C for 20 min. CONCLUSION: The fermented milk prepared with Lactobacillus helveticus 130B4 contained an ACE inhibitory peptide, κ‐CN 107–115. This fermented milk was expected to have anti‐hypertensive effect after ingestion because the peptide was stable to digestive protease and heat treatment in vitro. Copyright © 2008 Society of Chemical Industry     

Use of essential oil from Mentha arvensis L. to control storage moulds and insects in stored chickpea

BACKGROUND: Fungal contamination and Callosobruchus infestation results in qualitative and quantitative losses of chickpea seeds during storage. Most of the synthetic chemicals used as preservatives have adverse effects. Therefore, the antifungal and insecticidal potential of Mentha arvensis essential oil was evaluated to determine whether this could be an eco‐friendly substitute of synthetic preservatives. RESULTS: The stored chickpea seeds were dominated by Aspergillus flavus (46.1%) and 30% isolates among them were found toxigenic. The MIC of Mentha oil against A. flavus was recorded at 400 µL L^?1 and it exhibited broad fungitoxic activity against 14 storage fungi. The oil was found superior to some prevalent synthetic fungicides. Mentha oil showed potent insecticidal activity against Callosobruchus chinensis at different concentrations and exposure times. The oviposition by C. chinensis was completely checked at 10 µL L^?1 while F₁ emergence was completely inhibited at 200 µL L^?1. During in situ experiments, 94.05% protection of the chickpea from C. chinensis by Mentha oil showed superiority over the organophosphate insecticide malathion, where 90.75% protection was recorded. CONCLUSION: The Mentha EO showing potent fungitoxic and insecticidal efficacy and may be recommended as a plant‐based preservative in the management of fungal and insect infestation of chickpea and other pulses during storage. Copyright © 2009 Society of Chemical Industry     

Characterization of the recombinant succinic semi‐aldehyde dehydrogenase from Saccharomyces cerevisiae

The yeast succinic semi‐aldehyde dehydrogenase gene (SSADH; EC 1.2.1.16) was cloned and overexpressed in Escherichia coli. Based on SDS–PAGE, the molecular mass of the subunit was around 54 kDa, and the purified recombinant enzyme had a tetrameric molecular mass of ca. 200 kDa. The specific activity of the recombinant enzyme was 1.90 µm NADH formed/min/mg, and showed maximal activity at pH 8.4. The recombinant protein was highly specific for succinate semi‐aldehyde (K_m = 15.48 ± 0.14 µm ) and could use both NAD⁺ and NADP⁺ as co‐factors, with K_m values of 579.06 ± 30.1 µm and 1.017 ± 0.46 mm, respectively. Initial velocity studies showed that NADH was a competitive inhibitor with respect to NAD⁺ (K_i = 129.5 µm ) but a non‐competitive inhibitor with respect to succinate semi‐aldehyde. Adenine nucleotides of AMP, ADP and ATP inhibited yeast SSADH activity with K_i = 1.13–10.2 mm, and showed competitive inhibition with respect to NAD⁺ and mixed‐competitive, non‐competitive and non‐competitive inhibition, respectively, with respect to succinate semi‐aldehyde. The kinetic data suggest a 'ping‐pong' mechanism. Copyright © 2014 John Wiley & Sons, Ltd.     

Isolation of eriocitrin (eriodictyol 7‐O‐rutinoside) as an arachidonate lipoxygenase inhibitor from Lumie fruit (Citrus lumia) and its distribution in Citrus species

An inhibitory compound acting against rat platelet 12‐lipoxygenase was isolated from the peel of Lumie fruit (Citrus lumia) by activity‐guided separation. It was identified as eriocitrin (eriodictyol 7‐O‐rutinoside) by spectroscopic analyses. Eriocitrin inhibited 5‐lipoxygenase (IC₅₀29.1 µmol L^?1) from rat peritoneal polymorphonuclear leukocytes in addition to 12‐lipoxygenase (IC₅₀22.3 µmol L^?1). Its aglycone, eriodictyol (5,7,3′, 4′‐tetrahydroxyflavanone), was a much more potent inhibitor of both 12‐lipoxygenase (IC₅₀0.07 µmol L^?1) and 5‐lipoxygenase (IC₅₀0.20 µmol L^?1). It also inhibited the production of leukotriene B₄ in intact peritoneal polymorphonuclear leukocytes stimulated with calcium ionophore A23187 (IC₅₀12.7 µmol L^?1). The distribution of eriocitrin in 39 citrus fruits was investigated by high‐performance liquid chromatography analysis. Lumie, eureka lemon (Citrus limon), Sambokan (Citrus sulcata), Sudachi (Citrus sudachi) and Koji (Citrus leiocarpa) fruits were found to contain high levels of eriocitrin in both peel and juice vesicles. Copyright © 2006 Society of Chemical Industry     

Relative severity of fumonisin contamination of cereal crops in West Africa

Traditional and improved varieties of maize, pearl millet and sorghum were planted by small-scale farmers under the direction of the International Institute for Tropical Agriculture in two Nigerian agro-ecological zones: the Sudan Savanna and the Northern Guinea Savanna. Samples were collected for the determination of Fusarium infection and fumonisin (B₁, B₂ and B₃) contamination. A previous paper reported Aspergillus infection and aflatoxin contamination of these samples. Fusarium infection levels, measured by per cent kernels infected, were modest with mean levels for the above cereals of 16% ± 11% (SD), 12% ± 7% and 13% ± 16%, respectively. However, the Fusarium species recovered from maize were predominantly the fumonisin producers F. verticillioides and F. proliferatum, together making an infection rate of 15% ± 10%, whereas these species were present to a limited extent only in the other two cereals, 1% ± 1% for pearl millet and 2% ± 6% for sorghum. Fumonisin contamination was variable but reflected the diversity of Fusarium producers in these three cereals. Mean levels were 228 ± 579 µg kg^–1 (range < 5–2860 µg kg^–1) for maize, 18 ± 7 µg kg^–1 (range = 6–29 µg kg^–1) for pearl millet and 131 ± 270 µg kg^–1 (range < 5–1340 µg kg^–1) for sorghum. Together with previous results on aflatoxin, this study confirmed the co-occurrence of aflatoxins and fumonisins in maize as well as in the traditional African cereals, millet and sorghum (89% co-occurrence across all three cereals). The low fumonisin levels may be ascribed to the use of good agricultural practices. Of the Fusarium species present, those in maize consisted mainly of fumonisin producers, the opposite of what was observed in pearl millet and sorghum. It is concluded that replacement of maize by pearl millet and sorghum could improve food safety with regards to aflatoxin B and fumonisin B exposure.     

In vitro and in vivo antifungal properties of cysteine proteinase inhibitor from green kiwifruit

BACKGROUND: Higher plants possess several mechanisms of defense against plant pathogens. Proteins actively synthesized in response to those stresses are called defense‐related proteins which, among others, include certain protease inhibitors. It is of particular relevance to investigate plant natural defense mechanisms for pathogen control which include cystatins—specific inhibitors of cysteine proteases. RESULTS: In this study, a cysteine proteinase inhibitor (CPI), 11 kDa in size, was purified from green kiwifruit to homogeneity. Immuno‐tissue print results indicated that CPI is most abundant in the outer layer of pericarp, near the peel, and the inner most part of the pulp—sites where it could act as a natural barrier against pathogens entering the fruit. The purified protein (15 µmol L^?1) showed antifungal activity against two phytopathogenic fungi (Alternaria radicina and Botrytis cinerea) by inhibiting fungal spore germination. In vivo, CPI (10 µmol L^?1) was able to prevent artificial infection of apple and carrot with spore suspension of B. cinerea and A. radicina, respectively. It also exerted activity on both intracellular and fermentation fluid proteinases. CONCLUSION: Identification and characterization of plant defense molecules is the first step towards creation of improved methods for pathogen control based on naturally occurring molecules. Copyright © 2012 Society of Chemical Industry     

Evaluation of Antimicrobial,Antioxidant Activities,and Nutritional Values of Fermented Foxtail Millet Extracts by Lactobacillus paracasei Fn032

Solid-state bioprocessing of foxtail millet by Lactobacillus paracasei Fn032 is a biotechnological strategy to produce fermented foxtail millet meal with more beneficial components. The objective of this study was to evaluate the antioxidant, antimicrobial properties and nutritional values of water extracts from fermented foxtail millet flour and its bran with and without protease. Fermented foxtail millet flour with added protease extract showed higher scavenging ability on 1,1-diphenyl-2-picrylhydrazyl radicals as well as reducing power than fermented foxtail millet flour and fermented foxtail millet bran extracts. Both extracts, fermented foxtail millet flour and fermented foxtail millet flour with protease, showed significant (P < 0.05) effectiveness inhibition abilities on microbial growth when compared with fermented foxtail millet bran extracts. Amino acid profile revealed that fermented foxtail millet flour with protease, with relatively strongest antioxidant, antimicrobial activity, also had the highest total hydrophobic amino acids content (51.39%) and hydrophobic index (8.47 Kj/mol amino acid residue). Moreover, fermented foxtail millet flour with protease revealed the highest protein content, predicted protein efficiency ratio, and protein digestibility. Molecular weight of the whole extracts varied from 180–5000 Da. Based on the results obtained, fermented foxtail millet flour extracts were relatively effective in the antioxidant, antimicrobial properties assayed and might be potential biological values for application in food products.     

Purification and properties of cysteine protease from rhizomes of Curcuma longa (Linn.)

BACKGROUND: Turmeric rhizome (Curcuma domestica Linn.) contains proteases and has proteolytic activity. Curcumin from turmeric rhizomes has been used for healing manu ailments, including cancer have been used for healing many ailments, including cancer. The purpose of this study was to purify turmeric protease and to research their biochemical characteristics. RESULTS: Cysteine protease from C. domestica has been purified to homogeneity using acetone precipitation followed by preparatory native polyacrylamide gel electrophoresis (PAGE). This protocol resulted in six fold purification with 28% final recovery. The purified turmeric protease showed a prominent single peak and band on high‐performance liquid chromatography and sodium dodecyl sulfate–PAGE, respectively, and an estimated molecular weight of 43 KDa, and exhibited optimal activity between 37 and 60 °C. The protease activity of the turmeric protease was significantly inhibited by iodoacetic acid. The turmeric protease had higher alanine and glutamate content and cleaved synthetic peptides N‐Cbz‐Ile‐Pro and N‐Cbz‐Phe‐Leu in a time‐dependent manner. Peptide mass fingerprint using matrix‐assisted laser desorption/ionization–time of flight mass spectroscopy revealed peptide matches to proteasome subunit alpha type 3 of Oryza sativa ssp. japonica (Rice). The turmeric protease showed antifungal activity at 10 µg mL^?1 towards pathogens Pythium aphanidermatum, Trichoderma viride and Fusarium sp. CONCLUSION: Cysteine addition significantly activated turmeric protease. The protease inhibition test suggested that turmeric protease belonged to the cysteine type. The biochemical characteristics of turmeric protease described in this paper can provide useful information for potential end uses of turmeric protease for pharmaceutical industry applications such as therapeutics. Copyright © 2009 Society of Chemical Industry     

Potential hypoglycaemic effects of insoluble fibres isolated from foxtail millets [Setaria italica (L.) P. Beauvois]

Insoluble fibres were isolated from the two varieties of foxtail millet (white and yellow) grains and evaluated for their hypoglycaemic effects by in vitro studies. The hypoglycaemic effects of these fibres were compared with those of commercial soy insoluble fibre. The results revealed that minimum and maximum amounts of glucose were adsorbed on each sample at 10 and 200 μmol g^?1 glucose concentrations respectively, indicating that the glucose adsorption capacity (GAC) of the fibre materials was proportional to glucose concentration for all samples. There was significant (P < 0.05) difference among all the fibre materials in relation to their GAC values. In the case of the effects of the fibres on glucose diffusion, the millets' insoluble fibres performed better than that of the commercial soy insoluble fibres. The glucose dialysis retardation indexes at the end of the maximum dialysis time were 1.1%, 27.4% and 22.6% for soy bean insoluble fibre, white foxtail millet insoluble fibre and yellow foxtail millet insoluble fibre in that order. The study showed that hypoglycaemic effects of yellow and white foxtail millet fibres were comparable to the commercial soy insoluble fibre.     

ANTIFUNGAL,AFLATOXIN INHIBITORY AND FREE RADICAL‐SCAVENGING ACTIVITIES OF SOME MEDICINAL PLANTS EXTRACTS

ABSTRACT

The antifungal, aflatoxin inhibitory and antioxidant activity of methanol–aqueous extract (2:1) of 62 medicinal plants was explored. Based on the antifungal results, the extracts of 25 plants showed more than 50% antifungal activity and were further investigated for their aflatoxin inhibition and antioxidant properties. Methanol–aqueous extracts of Phyllanthus emblica and Terminalia chebula fruits caused 100% inhibition of aflatoxin production by the toxigenic strain of Aspergillus flavus in semisynthetic medium at 1 mg/mL. In addition, P. emblica (IC₅₀ = 4.1 µg/mL) and T. chebula (IC₅₀ = 6.9 µg/mL) fruits extracts exhibited strong antioxidant activity during the 2,2‐diphenyl‐1‐picrylhydrazyl radical‐scavenging assay in comparison with butylated hydroxytoluene (IC₅₀ = 8.1 µg/mL) and butylated hydroxyanisole (IC₅₀ = 6 µg/mL).

PRACTICAL APPLICATIONS

Based on the results of the present study, methanol–aqueous extracts of Phyllanthus emblica and Terminalia chebula, being endowed with strong antifungal, aflatoxin inhibitory and antioxidant activity, may be recommended as plant‐based preservatives for the enhancement of shelf life of food items and their protection from the undesirable harmful effects of molds, aflatoxin and free radical‐mediated damages.     

Quantitative analysis of the isoflavone content and biological growth of soybean (Glycine max L.) at elevated temperature,CO2 level and N application

Effects of ambient and elevated CO₂ levels (360 and 650 µmol mol^?1 respectively), ambient and high (5 °C above ambient) temperatures and their interactions with N application on soybean (Glycine max L.) were studied in 2001. Overall, total isoflavones in whole soybean seeds were highest (1383.0 µg g^?1) in the elevated CO₂ (AE) treatment without N application and lowest (414.1 µg g^?1) in the elevated temperature (EA) treatment with N application. Malonylgenistin (449.2 µg g^?1) and malonyldaidzin (435.9 µg g^?1) concentrations in the AE treatment without N application were highest among the 12 individual isoflavones, while aglycon and acetyl conjugates showed lower concentrations (below 10 µg g^?1) than glucoside and malonyl conjugates in all treatments. Overall, N application had no effect on total isoflavone concentration, while both temperature and CO₂ level had a higher effect on increasing isoflavones, including aglycon and acetyl conjugates (P = 0.001). In the biological growth analysis, total dry weight was highest (100.9 g) in the elevated temperature and CO₂ (EE) treatment with N application, while leaf area was more affected by CO₂ than by temperature and increased with N application. There were larger numbers of pods (99) and seeds (176) per plant in the EE treatment with N application, and generally the AE treatment showed a greater increase in 100‐seed weight (g per 100 seeds) and in pods and seeds per plant than other treatments. Overall, total dry weight was highly affected (P = 0.001) by three main factors, temperature, CO₂ and N application, but the interactions temperature × N and temperature × CO₂× N did not affect total dry weight. Also, total dry weight tended to increase with increasing numbers of pods (r² = 0.93***) and seeds (r² = 0.93***) and larger leaf area (r² = 0.85***). In addition, numbers of pods and seeds were significantly affected (P = 0.01–0.001) by temperature, CO₂ and temperature × CO₂. Generally, elevated CO₂ and temperature did not affect N, P and K concentrations in the seeds but did decrease the concentrations of Ca and Mg, which were increased in the AE treatment. Among the nutrients, Ca and Mg were highly correlated with temperature and CO₂ level. N concentration in the seeds increased with applied N and in particular showed a high increase with elevated temperature and ambient CO₂ (EA treatment). The variation in isoflavones was correlated with temperature (r² = ?0.70**) and CO₂ (r² = 0.67**), while N application was not correlated with isoflavone concentration. Also, Ca (r² = ?0.85***) and Mg (r² = ?0.57*) in the seeds were correlated with variation in isoflavones. This indicated that isoflavones were in higher concentrations under conditions of low temperature and increasing CO₂, which also resulted in low Ca and Mg concentrations in the seeds. The results of this study suggest that the long‐term adaptation of the soybean to growth at elevated CO₂ level and high temperature might potentially increase its isoflavone content, growth and yield. Copyright © 2005 Society of Chemical Industry     

挤压和发酵对小米多肽ACE抑制活性及抗氧化作用的影响

以小米为原料,制备具有血管紧张素转换酶(ACE)抑制作用的食源性活性多肽。采用挤压和发酵的方法对小米粉进行处理,研究其对小米蛋白消化率的影响,同时对在胃蛋白酶-胰酶水解条件下得到的小米多肽进行ACE抑制活性和抗氧化能力分析。结果表明,相比于原粉,挤压和发酵对小米多肽的ACE抑制活性和抗氧化能力都有显著的影响(P<0.01);挤压对小米蛋白消化率的影响较大(53.11%);挤压小米蛋白水解肽具有较强的ACE抑制活性(IC50=0.057 mg肽/mL)、DPPH自由基清除能力(10.99μmol/g肽)和铁离子还原能力(82.92μmol/g肽)。     

Determination of protein, fat, starch, and amino acids in foxtail millet [Setaria italica (L.) Beauv.] by Fourier transform near-infrared reflectance spectroscopy

Quantitative detection of protein, fat, starch, and amino acids in foxtail millet using Fourier transform near-infrared spectroscopy (NIRS) was investigated. Foxtail millet samples (n=259) were analyzed using NIRS. Spectral data were linearized with data from chemical analyses. Calibration models were established using a partial least-squares (PLS) algorithm with cross-validation. Optimized models were tested using external validation set samples with coefficients of determination in the external validation (R ² _val) of >0.90. Residual predictive deviation (RPD) values were nearly equal to or >2.5 for crude protein, alanine, aspartic acid, glutamic acid, isoleucine, leucine, and serine. However, for glycine, histidine, phenylalanine, proline, threonine, tyrosine, and valine, the R ² _val values were >0.83 and RPD values were nearly equal to or >2.0. For crude fat, total starch, arginine, and lysine, the R ² _val values were >0.70 and RPD values were >1.5. NIRS is a rapid determination tool for foxtail millet breeding, and for quality control.     

The influence of heat treatment of chickpea seeds on antioxidant and fibroblast growth‐stimulating activity of peptide fractions obtained from proteins digested under simulated gastrointestinal conditions

This study presents the effect of heat treatment of chickpea seeds on biological activity of peptides obtained by in vitro gastrointestinal digestion. The most significant antiradical activity against ABTS^+? expressed as IC₅₀ value was observed for 3.5‐ to 7‐kDa peptide fraction from TC hydrolysate (41.01 μg mL^?1). In turn, peptide fraction of 3.5–7.0 kDa obtained from raw chickpea seeds hydrolysate showed the highest antiradical activity against DPPH^? and Fe²⁺ chelating activity with IC₅₀ value of 20.94 and 52.53 μg mL^?1, respectively. The highest Cu²⁺ chelating activity was observed for peptides obtained from TC hydrolysate (IC₅₀ = 56.60 μg mL^?1). Peptide fraction <3.5 kDa from TC hydrolysate demonstrated the most significant reducing power (0.362 A₇₀₀/μg mL^?1). The peptide fraction of 3.5–7 kDa from TC hydrolysate also showed the highest fibroblast growth‐stimulating activity. These results indicated that the heat treatment process has no significant effect on antiradical activity against DPPH^? and Fe²⁺ chelating ability of peptides.