Amylolytic activity in stored potato tubers. 1. Estimation usingp-nitrophenyloligosaccharides

Summary Methods for the estimation of amylolytic activity are reviewed. A procedure for the routine extraction of amylolytic activity from freeze-dried powder prepared from potato tubers is described. The extraction medium is buffered at pH 7.0 and contains glycerol, dithiothreitol, calcium chloride and the non-ionic detergent, nonidet P-40. α-Amylase activity and exoamylolytic activity were estimated in crude extracts of potato tubers using the substrates, blockedp-nitrophenyl-maltoheptaoside andp-nitrophenyl-maltopentaoside respectively. These substrates are included in kits supplied by Biocon, (UK) Ltd to measure the α- and ?-amylase activity in cereals. The validity of using these kits for the determination of α- and ?-amylase activity in potato tuber tissue is discussed.     

Amylolytic activity in stored potato tubers. 2. The effect of low-temperature storage on the activities of α-and β-amylase and α-glucosidase in potato tubers

Summary Tubers of the potato cultivars Record, Wilja, Pentland Dell and Brodick (formerly clone 137371) were sampled before and after storage at either 4°C or 10°C. Reducing sugar content stayed constant during storage at 10°C in all four cultivars but rose greatly during the first 6–12 weeks of storage at 4°C in Record, Wilja and Pentland Dell but not in Brodick. Amylolytic activity was determined after 5 weeks storage using blockedp-nitrophenyl maltoheptaoside as substrate for α-amylase,p-nitrophenyl maltopentaoside as substrate for β-amylase, andp-nitrophenylglucopyranoside as substrate for α-glucosidase. The values obtained from tubers stored at 4°C were higher than those from tubers stored at 10°C, the differences being much less in Brodick than in the other three cultivars.     

Studies on Indian Green Leafy Vegetables for Their Antioxidant Activity

To identify the potential of green leafy vegetables (GLV) as antioxidants, methanolic extracts of Amaranthus sp., Centella asiatica, Murraya koenigii and Trigonella foenum graecum were studied for their antioxidant activity in different systems at multiple concentrations. Total antioxidant activity assessed by phosphomolybdenum method, free radical scavenging activity by 1,1-diphenly-2-picryl hydrazyl (DPPH), reducing power and ferrous ion chelating activity were determined. The GLV were analyzed for ascorbic acid, total and β-carotene and total polyphenol contents. The ascorbic acid, total carotene, β-carotene and total phenolic content (tannic acid equivalents) of the GLV ranged between 15.18–101.36, 34.78–64.51, 4.23–8.84 and 150.0–387.50 mg/100 g GLV, respectively. The extracts were found to have significantly different levels of antioxidant activities in the systems tested. The total antioxidant activity was highest in Murraya koenigii (2,691.78 μmol of ascorbic acid/g sample) and least in Centella asiatica (623.78 μmol of ascorbic acid/g sample). The extract concentration causing 50% inhibition of DPPH (IC₅₀) was determined (M. koenigii < C.asiatica < Amaranthus sp. < T. graecum). The maximum DPPH scavenging activity and reducing power was exhibited by Murraya koenigii. Multiple regression analysis showed that the relationship of total antioxidant activity, free radical scavenging activity, and reducing power with polyphenol and total and β-carotene was highly significant. Paper awarded the Young Scientist Award in Experimental Nutrition at the Young Scientist Award Session of the 39th Annual Meet of the Nutrition Society of India, Hyderabad, India on 15–16th Dec 2007.     

Effect of High β-Glucan Barley on Serum Cholesterol Concentrations and Visceral Fat Area in Japanese Men—A Randomized,Double-blinded,Placebo-controlled Trial

This study investigated whether the consumption of a diet in which high-β-glucan barley replaced rice would reduce the visceral fat area as well as the serum low-density lipoprotein cholesterol (LDL-C) and total cholesterol (TC) in hypercholesterolemic Japanese men. A randomized, double-blinded, placebo-controlled intervention study was conducted in 44 hypercholesterolemic Japanese men with a body mass index (BMI) >22 kg/m². The subjects were randomly assigned to groups consuming either rice (placebo group) or a mixture of rice and pearl barley with a high β-glucan content (test group, 7.0 g β-glucan per day) for 12 weeks. Blood samples were taken, and CT scan obtained before the trial and every four weeks during the trial. The pearl barley intake significantly reduced serum concentrations of LDL-C (P = 0.041) and TC (P = 0.037) during the trial. Significant differences between the test and placebo groups were found for the visceral fat (P = 0.039), BMI (P = 0.015), and waist circumference (P = 0.011) at the end point. The consumption of pearl barley with a high β-glucan content reduces not only LDL-C but also visceral fat area.     

茯砖茶不同萃取物对消化酶活性的影响

将茯砖茶依次用三氯甲烷、乙酸乙酯及正丁醇萃取后制备样品,测定各部分样品对α-淀粉酶及胰脂肪酶活性的影响。研究发现茯砖茶各部分样品对胰脂肪酶活性的影响程度不同,三氯甲烷层稍有抑制作用,活性只有对照的0.82倍,水提物、乙酸乙酯萃取层及正丁醇萃取层有显著的激活作用,正丁醇萃取层部分激活作用最明显,达2.54倍。茯砖茶各部分样品对α-淀粉酶活性均有激活作用,其中以乙酸乙酯萃取层部位的激活作用最强,激活倍数为4.09,其次是水提物,其激活倍数为2.99,水层的激活作用不显著。三氯甲烷萃取层抑制作用不显著,活性只有对照的0.88倍。乙酸乙酯萃取层及正丁醇萃取层部分经过HP-20树脂再分离后,各部分仍有较高的活性。     

Amylolysis of wheat starches. I. Digestion kinetics of starches with varying functional properties

The susceptibility of wheat (Triticum aestivum L.) starches to hydrolysis by pancreatic α-amylase in vitro was investigated using a series of 35 starches with slightly enriched amylose content within a narrow range (36–43%), but widely differing functional properties. After 2 h of incubation with α-amylase, native starch granules were digested to different extents, but there were no differences between any of the starches once they were gelatinized. Cooling the starch for 72 h at 4 °C after cooking reduced the susceptibility of all of the starches to enzymic digestion by a similar extent, whereas addition of monopalmitin decreased the digestibility of the starches that contained amylose, but did not affect the digestibility of waxy starches that were also included in the study. Amylopectin chain length distribution of partly digested starch granules displayed increased proportion of short and medium chains and decreased proportion of long chains in comparison to native granules. Separated large (A) and small (B) starch granules from three of the starches differed significantly in their susceptibility to in-vitro digestion. A predictive model of the susceptibility of starch in the different forms was developed from the physico-chemical and functional properties of the starches.     

Characteristics of two major lectins from mungbean (Vigna radiata) seeds

Two major lectins, MBL-I and MBL-II, were purified fromVigna radiata L. seeds using ion-exchange and gel filtration chromatography techniques. MBL-I was found to be a tetramer with native M.W. of 132 kDa and subunit M.W. of 33 kDa having α-galactosidase activity. MBL-II consisted of two monomeric lectins with M.W. of 94 kDa and 89 kDa which were associated mainly with β-galactosidase activity. Both MBL-I and MBL-II are D-galactose-specific lectins.     

Distribution of Enzymes in Wheat Flour Mill Streams

The distribution of alpha -amylase, protease, lipoxygenase, polyphenol oxidase and peroxidase in wheat roller flour mill streams was studied. Break flours had relatively less alpha -amylase and protease activity than reduction flours both on flour weight and a protein basis. Among the different flour streams, the 5^thand 6^threduction passage had the highest alpha -amylase activity, while the 4^threduction passage had the highest protease activity. The lipoxygenase activity was concentrated mostly in the last break and the reduction streams, whereas polyphenol oxidase activity was highest in break flour streams. Peroxidase activity was distributed unevenly among the different mill streams. The lipoxygenase, polyphenol oxidase and peroxidase were highly concentrated in different bran fractions. Except for protease, the other enzymes were concentrated in the «atta», a milling by-product comprising refined flour, bran and shorts; and are least active in semolina (farina).     

Variation of Beta -Amylase Activity in Barley as Affected by Cultivar and Environment and its Relation to Protein Content and Grain Weight

The cultivar and environmental variation of beta -amylase activity was studied using two barley cultivars with contrasting growth properties. There was a significant difference in beta -amylase activity between the two cultivars used, 92-11 being significantly higher than Xiumai 3. A significant variation in beta -amylase activity was detected between grains at different positions within a spike. The two cultivars showed the same pattern, with top grains showing the highest and bottom ones the lowest activities. The relative difference within a spike varied between the cultivars, with 92-11 being larger than Xiumai 3. Both seeding rate and timing of N application dramatically affected the beta -amylase activity. With N application at the booting stage, beta -amylase activity increased, mainly due to the significantly increased beta-amylase activity in the topmost grains. The bottom grains showed a lower response to timing of N application. The variation in protein content and grain weight between cultivars and among the various treatments was also examined. The possible influence of these factors on beta -amylase activity are discussed.     

A rapid method for the quantification of steroidal glycoalkaloids by reversed phase HPLC

Summary Methods were developed for clean-up of potato tuber extracts on solid phase extraction CN-cartridges and for analysis of steroidal glycoalkaloids by reversed phase HPLC. The alkaloids α-solanine, α-chaconine, α-solasonine and α-solamargine could be separated on C₈ or C₁₈ reversed phase columns using a mobile phase of acetonitrile/Tris-buffer (3∶2, v/v). The analysis time of tuber extracts containing these alkaloids was less then 4 min, if the pH of the mobile phase was adjusted to 7.4 and run at a flow rate of 0.5 ml/min.     

Inhibitory Activity of Cinnamon Bark Species and their Combination Effect with Acarbose against Intestinal α-glucosidase and Pancreatic α-amylase

Inhibition of α-glucosidase and pancreatic α-amylase is one of the therapeutic approaches for delaying carbohydrate digestion, resulting in reduced postprandial glucose. The aim of this study was to evaluate the phytochemical analysis and the inhibitory effect of various cinnamon bark species against intestinal α-glucosidase and pancreatic α-amylase. The results showed that the content of total phenolic, flavonoid, and condensed tannin ranged from 0.17 to 0.21 g gallic acid equivalent/g extract, from 48.85 to 65.52 mg quercetin equivalent/g extract, and from 0.12 to 0.15 g catechin equivalent/g extract, respectively. The HPLC fingerprints of each cinnamon species were established. Among cinnamon species, Thai cinnamon extract was the most potent inhibitor against the intestinal maltase with the IC₅₀ values of 0.58 ± 0.01 mg/ml. The findings also showed that Ceylon cinnamon was the most effective intestinal sucrase and pancreatic α-amylase inhibitor with the IC₅₀ values of 0.42 ± 0.02 and 1.23 ± 0.02 mg/ml, respectively. In addition, cinnamon extracts produced additive inhibition against intestinal α-glucosidase and pancreatic α-amylase when combined with acarbose. These results suggest that cinnamon bark extracts may be potentially useful for the control of postprandial glucose in diabetic patients through inhibition of intestinal α-glucosidase and pancreatic α-amylase.     

Mechanisms Leading to Excess Alpha -Amylase Activity in Wheat (Triticum aestivum, L) Grain in the U.K.

The frequency and mechanisms of four modes of alpha -amylase enzyme accumulation in U.K. wheat, retained pericarp alpha -amylase activity (RPAA), pre-maturity alpha -amylase activity (PMAA), pre-maturity sprouting (PrMS) and post-maturity sprouting (PoMS), were investigated in field and laboratory experiments. Of 56 cultivar site year combinations (four model cultivars grown at up to four sites from 1994–1997), enzyme activity was detected in 32 cases, in 23 cases sufficient to reduce Hagberg falling number (the usual industry measure of alpha -amylase) below the commercial criterion (250 s). The frequency of occurrence of different modes of enzyme accumulation was in the order PoMS>PMAA>PrMS>RPAA. Both PMAA and PrMS were more common than expected and the most usual pattern was for alpha -amylase to accumulate by several modes. Although green grains are rejected as impurities, study of grain colour in relation to pericarp alpha -amylase activity showed that the enzyme could persist in non-green grains in levels sufficient to affect the Hagberg value. Two factors thought to promote PMAA, grain drying rate and transient changes in temperature in early development, were studied in the field and controlled environment cabinets. No significant difference was found in grain drying rate between samples where PMAA was or was not identified. However, out of 19 transfers from a cool (16/10 °C) to a warm (26/20 °C) temperature regime, six led to significant increases in PMAA. No transfers after 45% grain moisture increased PMAA. PrMS occurred as early as 67% grain moisture and susceptibility usually increased with stage of development, being greatest in the grain dough stage. PrMS susceptibility varied with cultivar (in the same order as PoMS sensitivity) and was affected by environmental factors.     

Hypocholesterolemic Activity from the Leaf Extracts of Cnidoscolus chayamansa

The aim of this study was to determine the hypocholesterolemic activity of Cnidoscolus chayamansa. In an in vivo model, high-cholesterol diet administered to mice Balb/c induced hypercholesterolemia. Three extracts from Cnidoscolus chayamansa (ethanol, methanol and an aqueous extract) were tested on hypercholesterolemic mice. Active extracts were assessed against the in vitro inhibitory activity of the same three extracts on the HMG-CoA reductase enzyme by using Vero cells. The specific chemical groups present in the phytochemical extracts were also determined. Only the aqueous extract (at either doses employed) showed a significant cholesterol reduction (27.9 and 31.1%, for 50 and 100 mg kg⁻¹, respectively P < 0.01). The extract did not inhibit the HMG-CoA reductase enzyme, suggesting that its compounds act at another level in cholesterol metabolism. Reactions to secondary metabolites indicate the presence of alkaloids in the aqueous and ethanol extracts and phenol hydroxyls in the ethanol and methanol extracts.     

Residues of free and bound maleic hydrazide in potato tubers

Summary The method of Vadukul (1991) for determining maleic hydrazide (MH) was modified and gave recoveries of free MH of 89%±4%. The values recorded on individual tubers ranged from 2–14 mg kg⁻¹. Maleic hydrazide was evenly distributed throughout the tuber (peel, outer and inner flesh) but concentration increased slightly as tuber size increased. The concentration of free MH decreased from 7 to 3 mg kg⁻¹ over the storage period of 5 1/2 months. Acid hydrolysis released substantial amounts of MH particularly from older potatoes (13 mg kg⁻¹) compared with 6 mg kg⁻¹ from new potatoes, implying that free MH is gradually converted to a bound form with time after treatment. No evidence was found for the presence of a β-glucoside of MH.     

Effects of Pressurized Low Polarity Water Extraction Parameters on Antioxidant Properties and Composition of Cow Cockle Seed Extracts

Antioxidant activity of pressurized low polarity water (PLPW) extracts of cow cockle seed and extraction residues were determined using DPPH, ABTS, and FRAP assays. The effect of extraction conditions (temperature (125, 150 and 175 °C) and time) on the antioxidant activity and the relationship amongst the antioxidant activity and extract composition (total phenolics and saponin content) were determined. The antioxidant activity of PLPW extracts increased with extraction temperature. Increasing activity with time was also observed at 175 °C. PLPW extraction residues had the highest activity suggesting antioxidant compounds were not completely extracted by PLPW. Antioxidant activity correlated well with total phenolics content of samples (R ² ≥ 0.94), however no correlation was observed with the saponin content. A strong correlation was observed between the antioxidant activity values obtained using different methods (R ² ≥ 0.94). These results point to the potential of PLPW extraction as a method to modify the activity of biological materials for the production of customized extracts.     

Production of D-lactic acid by bacterial fermentation of rice

This study describes the synthesis of D-lactic acid from rice by fermentation using microorganisms. Some micro-organisms were found to be active for producing D-lactic acid of high optical purity after an intensive screening test for D-lactic acid bacteria using glucose as substrate. Rice powder was hydrolyzed with a combination of enzymes:α-amylase,β-amylase, and pullulanase, and the resultant rice saccharificate was subjected to the fermentation with the selected D-lactic acid bacteria. After the optimization of this fermentation it has been confirmed that D-lactic acid can be manufactured in a pilot scale.     

Optimization of High-Protein Glutinous Rice Flour Production Using Response Surface Method

A response surface method was employed to study the effect of α-amylase concentration, hydrolysis temperature and time on the production of high protein glutinous rice flour (HPGRF). The suspension of glutinous rice flour (15%) that contained 6.52% protein was gelatinized and subsequently hydrolyzed by thermostable α-amylase. The hydrolysis yielded 0.144–0.222 g/g HPGRF with 29.4%–45.4% protein content. Hydrolysis time exerted a significant effect, while enzyme concentration and hydrolysis temperature showed insignificant effect on the protein content and production yield of HPGRF. The result of response surface method showed that the optimum condition for the production of HPGRF that contained at least 36% protein was treating gelatinized 15% glutinous rice flour suspension with 0.90 Kilo Novo α-amylase Unit (KNU)/g α-amylase at 80 °C for 99 min. By carrying out the predicted hydrolysis condition, HPGRF with 35.9% protein and 61.8% carbohydrates was resulted. The process yielded 0.172 g/g HPGRF. HPGRF contained higher amount of essential amino acids compared to glutinous rice flour. HPGRF had higher solubility and lower swelling power, and also showed no pasting peak compared with glutinous rice flour.     

Increased Level of Tetrahydro-β-Carboline Derivatives in Short-Term Fermented Garlic

In our previous study [1], we found that relatively short-term spontaneous fermentation (40 days at 60–70°C, 85–95% relative humidity) potentiates anti-oxidative properties of garlic, in which scavenging activity against hydrogen peroxide was included. Since tetrahydro-β-carboline derivatives (THβCs) that possess hydrogen peroxide scavenging activity have recently been identified in aged garlic extract, THβCs were quantitatively analyzed with liquid chromatography-mass spectrometry (LC-MS). (1R, 3S)-1-Methyl-1,2,3,4-tetrahydro-β-carboline-3-carboxylic acid (MTCC) and (1S, 3S)-MTCC were found in the fermented garlic extract whereas only trace levels of MTCCs were detected in the row garlic extract. Therefore, it is suggested that relatively short-term fermentation potentiates scavenging activity of garlic against hydrogen peroxide by forming THβCs, especially MTCCs.     

Determination of the Antimicrobial and Antioxidative Properties and Total Phenolics of Two “Endemic” Lamiaceae Species from Turkey: <Emphasis Type="Italic">Ballota rotundifolia</Emphasis> L. and <Emphasis Type="Italic">Teucrium chamaedrys</Emphasis> C. Koch

This study was designed to examine the in vitro antimicrobial and antioxidant activities and the amount of total phenolics of the methanol extracts of Ballota rotundifolia L. and Teucrium chamaedrys C. Koch. In the case of antimicrobial activity tests, polar sub-fractions of the methanol extracts of plant species exhibited weakest antimicrobial activity when compared with the non-polar ones. While, non-polar sub-fraction of B. rotundifolia showed moderate activity against A. lwoffii, C. perfringens and the yeasts, T. chamaedrys performed excellent activity pattern against all of the tested microorganisms. The sub-fractions were also screened for their possible antioxidant activities by two complementary tests, namely DPPH free radical-scavenging and β-carotene/linoleic acid assays. Non-polar extracts of the plant species remained inactive in both test systems. On the other hand polar extracts showed remarkable antioxidant activities. In DPPH system, free radical scavenging effect of T. chamaedrys was measured as 18.00 ± 1.42 μg.mg⁻¹. It is extremely important to point out that, polar sub-fraction of T. chamaedrys is found as effective as the positive control BHT. Non-polar sub-fraction of T. chamaedrys found to have the highest total phenolic amount (97.12 ± 1.28 μg/mg). Results obtained from this experiment confirm the relationship between the amount of phenolics and biological activities.     

Effect of home processing and storage on ascorbic acid and β-carotene content of bathua (Chenopodium album) and fenugreek (Trigonella foenum graecum) leaves

The present investigation was conducted to study the effect of selected processing and storage methods on the concentration of ascorbic acid and β-carotene in Bathua and fenugreek, leaves. Methods included storage of leaves with or without polythene bags for 24 and 48 h in a refrigerator at 5°C; at 30°C in polythene bags; drying (sun and oven); blanching (5, 10, 15 min); open pan and pressure cooking. Ascorbic acid content of fresh leaves was 220.97 to 377.65 mg and β-carotene content was 19.00 to 24.64 mg/100 g, DW. The percent loss of ascorbic acid ranged from 2.03 to 8.77 and 45.15 to 66.9 while lower losses (0.0 to 1.75 and 1.63 to 2.84) of β-carotene were observed in leaves, stored in, the refrigerator and at 30°C, respectively. A markedly greater reduction in ascorbic acid and β-carotene was observed in dried, blanched and cooked leaves. The study data suggest that storage of leaves in refrigeration, drying in oven, blanching for a short time and cooking in a pressure cooker results in better retention of these two vitamins.