普鲁兰酶N467G突变体的酶学性质分析

在淀粉制糖工业中,普鲁兰酶通常与糖化酶配合使用,其在酸性pH和较高温度下的催化活力是影响淀粉脱支效率的主要因素.该研究通过对长野芽胞杆菌(Bacillus naganoensis)普鲁兰酶蛋白质解折叠自由能的差值(ΔΔG)的计算预测和突变位点稳定性分析,选择突变位点并进行定点突变,获得突变体N467G.与野生型普鲁兰酶...     

Antioxidant activities of extracts from selected culinary herbs and spices

Recently, interest in plant-derived food additives has grown, mainly because synthetic antioxidants suffer from several drawbacks. Furthermore, plant extracts have been shown to possess health-promoting properties. In the present study, hydrodistilled extracts from basil, laurel, parsley, juniper, aniseed, fennel, cumin, cardamom, and ginger were assessed for their total phenol content, and antioxidant (iron(III) reduction, inhibition of linoleic acid peroxidation, iron(II) chelation, 1,1-diphenyl-2-picrylhydrazyl radical-scavenging and inhibition of hydroxyl radical-mediated 2-deoxy-d-ribose degradation, site and nonsite-specific) activities. The extracts from basil and laurel possessed the highest antioxidant activities except for iron chelation. Although parsley showed the best performance in the iron chelation assay, it was less effective at retarding the oxidation of linoleic acid. In the linoleic acid peroxidation assay, 1 g of the basil and laurel extracts were as effective as 177 and 212 mg of trolox, respectively. Thus, both extracts are promising alternatives to synthetic substances as food ingredients with antioxidant activity.     

Thermal inactivation kinetics of peroxidase and lipoxygenase from fresh pinto beans (Phaseolus vulgaris)

 Thermal inactivation kinetics of crude peroxidase (POX) and lipoxygenase (LOX) in fresh pinto beans were studied over the temperature range of 55–90°C. The inactivation of both enzymes followed first-order kinetics. The biphasic inactivation curves for POX indicate the existence of several isoenzymes of varying heat stability. In the temperature range of 55–70°C, the activation energies (E _a) of POX were 46.5 kcal·mol^–1 for the heat-labile portion and 37.6 kcal·mol^–1 for the heat-stable portion. On the other hand, the LOX enzyme had an E _a value of 42.26 kcal·mol^–1 at 55–75°C and 49.1 kcal·mol^–1 at 55–90°C. Received: 28 July 1997 / Revised version: 16 October 1997     

Thermal inactivation kinetics of peroxidase and lipoxygenase from fresh pinto beans (Phaseolus vulgaris)

 Thermal inactivation kinetics of crude peroxidase (POX) and lipoxygenase (LOX) in fresh pinto beans were studied over the temperature range of 55–90°C. The inactivation of both enzymes followed first-order kinetics. The biphasic inactivation curves for POX indicate the existence of several isoenzymes of varying heat stability. In the temperature range of 55–70°C, the activation energies (E _a) of POX were 46.5 kcal·mol^–1 for the heat-labile portion and 37.6 kcal·mol^–1 for the heat-stable portion. On the other hand, the LOX enzyme had an E _a value of 42.26 kcal·mol^–1 at 55–75°C and 49.1 kcal·mol^–1 at 55–90°C. Received: 28 July 1997 / Revised version: 16 October 1997     

Fates of seeded Escherichia coli O157:H7 and Salmonella on selected fresh culinary herbs during refrigerated storage

The fates of seeded Escherichia coli O157:H7 and Salmonella on selected fresh culinary herbs were evaluated at a refrigerated temperature (4 degrees C). Fresh herbs, including cilantro, oregano, basil, chive, parsley, and rosemary, were inoculated with six-strain mixtures of E. coli O157:H7 and Salmonella, and the microbial populations were monitored at 1, 5, 11, 16, 19, and 24 days. For both pathogens, a significant decrease in the population (P < 0.0001) occurred within the first 5 days of storage (< 0.8 log). Both pathogens remained the highest on cilantro and the lowest on rosemary (P < 0.0001). Storage time had a significant effect on the survival of E. coli O157:H7; populations declined as storage time progressed. Although storage of cilantro, basil, and chive was terminated after 19 days because of deteriorated quality, significant numbers of both pathogens were recovered from the remaining fresh herbs after 24 days of storage. The results showed that both bacteria were extremely persistent on all test herbs under the test conditions. The results also reinforce the concept that, once contaminated, bacterial pathogens can persist on fresh herbs throughout a normal distribution time.     

High pressure inactivation of lipoxygenase in soy milk and crude soybean extract

The high pressure inactivation of lipoxygenase (LOX) in soy milk and crude soybean extract was studied in the pressure range 0.1–650 MPa with temperature varying from 5 to 60 °C. For both systems, the isobaric–isothermal inactivation of LOX was irreversible and followed a first-order reaction at all pressure–temperature combinations tested. In the entire pressure–temperature area studied, the LOX inactivation rate constants increased with increasing pressure at constant temperature for both systems; the rate constants were somewhat smaller in soy milk system than in crude soybean extract. At constant elevated pressure, LOX exhibited the greatest stability around 20 °C in both systems, indicating that the Arrhenius equation was not valid over the entire temperature range. For both systems, the temperature dependence of the LOX inactivation rate constants at high temperature decreased with increasing pressure, while the highest sensitivity of the LOX inactivation rate constants to pressure was observed at about 30 °C. The pressure–temperature dependence of the LOX inactivation rate constants was successfully described either using an empirical mathematical model or using a thermodynamic kinetic model for both systems. On a kinetic basis, neither the reaction order of inactivation nor the pressure and temperature sensitivities of the inactivation rate constants were influenced by the different levels of food complexity between the two systems.     

烟叶在烘烤过程中脂氧合酶、脱落酸与色素降解的关系

随着烘烤变黄过程的进展,烟叶脂氧合酶活性、超氧自由基含量和脱落酸含量在0~24h缓慢增加,24~48h急剧增加达到高峰,然后开始下降。不同环境湿度条件下,变化规律相同,但变化速度有差异。低湿变黄变化迅速,高峰出现的时间早,峰值比较高,高湿变化速度慢,峰值也较低。脂氧合酶活性与脱落酸含量及叶绿素、类胡萝卜素的降解速度均呈正相关。      

Combined thermal and high pressure inactivation kinetics of tomato lipoxygenase

The combined isothermal (10–60 °C) and isobaric (0.1–650 MPa) inactivation kinetics of lipoxygenase (LOX) extracted from tomatoes and reconstituted in a tomato purée were studied. Thermal inactivation of LOX at atmospheric pressure proceeded in the temperature range of 45–65 °C. LOX inactivation did not follow first order kinetics; the data could be fitted assuming that the two isoforms of LOX with different thermostability were present. Combined thermal and high pressure inactivation occurs at pressures in the range of 100–650 MPa combined with temperatures from 10–60 °C, and followed first-order kinetics. In the high-temperature/low-pressure range, (T≥50 °C and P≤300 MPa) an antagonistic effect is observed, therefore, the Arrhenius and Eyring equation cannot be used over the entire temperature and pressure range. Small temperature dependence is found in the low-temperature/high pressure range. A third degree polynomial model was successfully applied to describe the temperature–pressure dependence of the inactivation rate constants, which can be useful to predict inactivation rate constants of tomato LOX reconstituted in tomato purée in the temperature–pressure range studied.     

Kinetic parameters of surface color degradation of canned fresh green peas sterilized in a rotary retort

The kinetic parameters of surface color degradation of canned fresh green peas sterilized in a rotary retort, using the a* value from a spectrophotometer as the physical parameter and the concept of fractional conversion in conditions of unsteady state were determined in the range of retort temperatures of 120 and 131 °C. Experimental initial a* values fluctuated between −14.4 and −15.7, while final a* values ranged between −0.165 and −4.75. After five iterations performed, solving simultaneously the energy balance of the system, the differential equation of unsteady conduction for spheres and the first-order degradation reaction of surface color by numerical methods, the best values of k_100 °C and E_a determined were 5.48×10⁻⁴ s⁻¹ and 89.37 kJ/mol respectively, being 5.68% the deviation between experimental and predicted fractional retentions of a* with the determined parameters.     

Crude lipoxygenase from pig muscle: partial characterization and interactions of temperature, NaCl and pH on its activity

Crude lipoxygenase (LOX) was extracted from fresh pig bacon belly and studied of partial characteristics. The interactions of temperature, sodium chloride (NaCl) and pH on LOX activity were investigated by response surface methodology (RSM). Kinetic studies indicated that the Michaelis constant (K(m)) and maximum velocity (V(max)) for LOX activity using linoleic acid as substrate were 68 μM and 0.26 U/min at 20°C, respectively. The optimal conditions for this reaction were: substrate concentration 3.47 mM, reaction temperature 39°C and pH ≧9.0. The NaCl critical value for LOX activity was 3.0% (w/w) at 20°C, above which the LOX activity began to decrease. Temperature had significant interactions (p<0.05) with NaCl and pH. The temperature critical value decreased with NaCl content increasing, while increased with pH increasing. These indicated that LOX activity and enzyme-catalyzed lipid oxidation in dry-cured meat products could be regulated by controlling process factors during the processing.     

Combined effect of temperature and pulsed electric fields on soya milk lipoxygenase inactivation

Pulsed electric fields (PEF) were applied to freshly prepared soya milk using a laboratory scale continuous PEF system to study the feasibility of inactivating lipoxygenase (LOX). Square wave PEF using different combinations of pre-treatment temperature, electric field strength and treatment time were evaluated in this study. Inactivation curves for the enzyme were plotted for each parameter and inactivation kinetics were calculated and modelled. Results showed the highest level of inactivation (84.5%) was obtained using a combination of preheating to 50 °C, and a PEF treatment time of 100 μs at 40 kV/cm. Inactivation of LOX activity as a function of treatment time could be described using a first order kinetic model. Calculated D values following pre-heating to 50 °C were 172.9, 141.6 and 126.1 μs at 20, 30 and 40 kV/cm, respectively.     

Impact of high-intensity pulsed electric field variables affecting peroxidase and lipoxygenase activities of watermelon juice

The effects of pulse frequency (50-250 Hz), pulse width (1.0-7.0 μs) and polarity (monopolar or bipolar) of high-intensity pulsed electric field (HIPEF) treatments (35 kV/cm and 1000 μs total treatment time) on peroxidase (POD) and lipoxygenase (LOX) activities were evaluated using a response surface methodology. Second-order expressions were accurate enough to fit experimental results. HIPEF bipolar treatments resulted to be more effective than monopolar treatments in reducing POD and LOX activities. Watermelon juice LOX was more resistant to HIPEF than POD within the range of assayed conditions. HIPEF treatments set at 50 Hz for 1.0 μs pulse width could attain minimum residual POD activity values up to 10%, whereas the highest POD activity reduction was reached by combining high frequencies and pulse widths. Thus, POD could be totally inactivated by applying 7.0-μs bipolar pulses at frequencies higher than 114 Hz. In addition, the effect of frequency on the LOX activity was highly affected by the pulse width of the treatment. Thus, treatments conducted at 220 Hz with bipolar pulses of 1.0 μs led to the lowest residual LOX activity (50%).     

Anti-tumorigenic activity of five culinary and medicinal herbs grown under greenhouse conditions and their combination effects

BACKGROUND: Herbs and spices have been used as food preservatives, flavorings, and in traditional medicines for thousands of years. More and more scientific evidence supports the medicinal properties of culinary herbs. Colon cancer is the third leading cause of cancer death in the USA, and the fourth most common form of cancer worldwide. The objectives of this study were to evaluate the antitumor activity of five selected herbs grown under greenhouse conditions, and to study the potential synergistic effects among different herbal extract combinations. RESULTS: Thyme, rosemary, sage, spearmint, and peppermint extracts significantly inhibited SW‐480 colon cancer cell growth, with sage extracts exhibiting the highest bioactivity, with 50% inhibition at 35.9 µg mL^?1, which was equivalent to 93.9 µg dried leaves mL^?1 of culture medium. Some mixtures of different herbal extracts had combination effects on cancer cell growth. The inhibitory effects of peppermint + sage combinations at a 1:1 ratio were significantly higher than rosemary + sage combinations at 1:1 ratio, although peppermint extracts showed lower inhibition than rosemary extracts. CONCLUSION: Extracts from herb species (thyme, rosemary, sage, spearmint and peppermint) can significantly inhibit the growth of human colon cancer cells. Mixtures of herb extracts can have combination effects on cancer cell growth. The study suggests that these five herbs may have potential health benefits to suppress colon cancer. Copyright © 2011 Society of Chemical Industry     

Effect of lipoxygenase activity in defatted soybean flour on the gelling properties of soybean protein isolate

Soybean lipoxygenase was inactivated to different degrees by dry heating of defatted soybean flour for 0, 5, 10, 15, 20 and 25 min and soy protein isolates were prepared thereof by isoelectric precipitation of the water extract of the defatted soybean flour. The fluorescence emission intensity at 420 nm of the chloroform–methanol extract of soy protein isolates, which was indicator of the existence of peroxidized lipid, varied in parallel with the lipoxygenase residual activity in defatted soybean flours. The dispersion of soy protein isolate showed an increasing turbidity with the increase of lipoxygenase residual activity in the starting defatted soybean flour, suggesting an elevated tendency to form insoluble aggregates during the preparation of soy protein isolate. Small deformation rheological test revealed that the gelling times were shorter for those soy protein isolates derived from low lipoxygenase activity defatted soybean flours than that of high lipoxygenase activity. Frequency sweep showed that G′ of soy protein isolate derived from low lipoxygenase defatted soybean flour was independent of oscillation frequency in contrast to that of derived from non dry-heated defatted soybean flour, the latter showed a marked frequency dependence. Large deformation test revealed that the gel hardness increased about 10 times after dry heating of defatted soybean flour for 20 min. As the increase of the lipoxygenase residual activity, the gel permeability increased markedly, suggesting that soy protein isolate from high lipoxygenase defatted soybean flour produced coarser textured gel, which corresponded well with the results of scanning electron microscopy.     

An assessment of dietary supplementation with tea catechins and rosemary extract on the quality of fresh beef

The effect of supplementation of beef cattle diets with tea catechins (TC) (1000mg/animal/day) and rosemary extract (RE) (1000mg/animal/day), for 103 days preceding slaughter, on the oxidative stability of M. longissimus dorsi (LD) steaks was evaluated. Dietary supplementation with TC and RE did not increase plasma total antioxidant status (TAS), LD α-tocopherol concentrations or pH. In LD steaks stored aerobically or in modified atmosphere packs (80% O(2):20% CO(2)) (MAP) for up to 8 days at 4°C, surface redness (CIE 'a' redness value) and lipid stability (TBARS, mg MDA (malondialdehyde)/kg muscle) were not significantly improved as a result of supplementation with TC and RE. Similarly no improvement in the sensory properties and lipid stability of cooked LD slices, stored aerobically or in 30% CO(2):70% N(2) for up to 11 days at 4°C, was observed. An in vitro fermentation study demonstrated that TC and RE were not fermented to any great extent under simulated rumen conditions. Direct addition of TC (1000ppm) and RE (1000ppm) significantly (P<0.05) improved the colour and lipid stability in LD patties stored in 80% O(2):20% CO(2) for up to 8 days at 4°C, thus, demonstrating the antioxidant potential of TC and RE supplements employed in the present study.     

Composition of flavonoids in fresh herbs and calculation of flavonoid intake by use of herbs in traditional Danish dishes

Many herbs are known as excellent sources of natural antioxidants, and consumption of fresh herbs in the diet may therefore contribute to the daily antioxidant intake. The present study was performed in order to quantify flavonoids in commonly eaten fresh herbs. Fifteen fresh herbs (basil, chives, coriander, cress, dill, lemon balm, lovage, oregano, parsley, rosemary, sage, spearmint, tarragon, thyme, and watercress) were analysed by HPLC and mass spectrometry. Five major flavonoid aglycones were detected and quantified by HPLC after acid hydrolysis: apigenin, isorhamnetin, kaempferol, luteolin, and quercetin. The highest levels of flavonoids were found in parsley (510–630 mg apigenin /100 g), lovage (170 mg quercetin/100g), mint (18–100 mg apigenin/100 g), and dill (48–110 mg quercetin/100 g). Mass spectrometric detection, using atmospheric pressure chemical ionisation (APCI), was used to verify the presence of flavonoids in the hydrolysed extracts of herbs. Some traditional Danish dishes contain herbs, particularly parsley, dill, cress and chives, and the contribution to the flavonoid intake by consumption of these dishes was calculated.     

Salmonella Typhimurium internalization is variable in leafy vegetables and fresh herbs

Despite washing and decontamination, outbreaks linked to consumption of fresh or minimally-processed leafy greens have been increasingly reported in recent years. In order to assure the safety of produce it is necessary to gain knowledge regarding the exact routes of contamination. Leaf internalization through stomata was previously reported as a potential route of contamination, which renders food-borne pathogens protected from washing and disinfection by sanitizers. In the present study we have examined the incidence (percentage of microscopic fields harboring ≥ 1 GFP-tagged bacteria) of Salmonella Typhimurium on the surface and underneath the epidermis in detached leaves of seven vegetables and fresh herbs. The incidence of internalized Salmonella varied considerably among the different plants. The highest incidence was observed in iceberg lettuce (81 ± 16%) and arugula leaves (88 ± 16%), while romaine (16 ± 16%) and red-lettuce (20 ± 15%), showed significantly lower incidence (P < 0.05). Internalization incidence in fresh basil was 46 ± 12%, while parsley and tomato leaves demonstrated only marginal internalization (1.9 ± 3.3% and 0.56 ± 1.36%, respectively). Internalization of Salmonella in iceberg lettuce largely varied (0-100%) through a 2 year survey, with a higher incidence occurring mainly in the summer. These results imply that Salmonella internalization occurs in several leafy vegetables and fresh herbs, other than iceberg lettuce, yet the level of internalization largely varies among plants and within the same crop. Since internalized bacteria may evade disinfection, it is of great interest to identify plants which are more susceptible to bacterial internalization, as well as plant and environmental factors that affect internalization.     

冰鲜鸡中主要腐败菌的分离鉴定及其产生物胺特性分析

本文用不同选择性培养基对冰鲜鸡的主要腐败菌进行分离,利用16S r RNA基因序列测定法对分离菌进行鉴定,最后利用薄层层析和HPLC法对分离菌是否代谢产生物胺进行分析。研究发现:从冰鲜鸡中共分离筛选得到11株腐败菌,分别命名为C1、C4、C7、C8、PCA1、PCA2、PCA3、PCA4、PCA5、PCA7、PCA8。PCA1与Stentrophomona(狭长平胞属)、PCA4与Kouria(考克氏菌属)、PCA3与Enterobacter asburiae(阿氏肠杆菌)、PCA7与Enterobacter cancerogenus(生癌肠杆菌)、C1与Enterobacter ludwigii(肠杆菌属)、C4与Enterobacter cloacae(阴沟肠杆菌)、C7与Enterobacter hormaechei(肠杆菌属)、C8与Enterobacter hormaechei(肠杆菌属)、PCA2与Kocuria rhizophila(克氏库克菌属)、PCA5和PCA8与Staphylococcus(葡萄球菌属)亲缘关系近。通过薄层层析初步鉴定菌是否有产生物胺能力,发现PCA2、PCA5、PCA8不产生物胺,其余菌均产生物胺。利用HPLC进一步确定产生物胺菌发酵液中生物胺组成和含量,结果 PCA7只产生腐胺,含量为844.937μg/m L;其余菌株都产含量不同的腐胺和尸胺。本文研究表明,冰鲜鸡易受肠道菌污染,且产腐胺与尸胺,为冰鲜鸡的防腐与安全食用提供一定的参考依据。      

Biochemical,biological and histological evaluation of some culinary and medicinal herbs grown under greenhouse and field conditions

BACKGROUND: Increasing evidence supports the potential health benefits of herbal extracts displaying antioxidant, anti‐inflammatory and antitumour activities. Environment can have a pronounced effect on phenolic content and antioxidant capacity. The objectives of this study were to evaluate the total phenolic contents and antioxidant capacities of five different herbs grown under greenhouse and field conditions and to assess their potential anti‐inflammatory effects. RESULTS: High total polyphenolic (TPP) content (measured by the Folin‐Ciocalteu reagent method) and high Trolox equivalent antioxidant capacity (TEAC) were observed in all herbs evaluated. Leaves from thyme, sage, spearmint and peppermint grown in the greenhouse showed significantly higher TPP content and TEAC than those grown under field conditions, with a threefold difference being observed in peppermint. Rosemary, spearmint and peppermint extracts showed stronger inhibition of cyclooxygenase COX‐2 than of COX‐1. CONCLUSION: The results show that producing herbs under greenhouse conditions can improve their biological activities by increasing TPP contents and antioxidant capacities. The selective inhibition of COX‐2 activity by rosemary, spearmint and peppermint suggests that they may be useful as anti‐inflammatory agents with fewer side effects than regular non‐steroidal drugs. Copyright © 2010 Society of Chemical Industry     

Nitrate in leafy vegetables,culinary herbs,and cucumber grown under cover in Estonia: content and intake

The content of nitrate in leafy vegetables, culinary herbs, and cucumber was determined during the years 2006–2008. All samples of Estonian origin, except white cabbage, were grown under cover. Seasonal differences in nitrate concentrations were observed in lettuce and spinach. Nitrate concentrations in lettuce were 22% and those in spinach were 24% higher in winter crops compared with samples collected in summer. The mean nitrate level was 3023 mg kg^–1 for fresh lettuce and 2337 mg kg^–1 for spinach. On average, 11.6% of fresh lettuce and spinach samples nitrate concentration exceeded the maximum level specified in European Commission Regulation No. 1881/2006. The mean levels were 999 mg kg^–1 for imported iceberg lettuce and 1287 mg kg^–1 for frozen spinach, which are below the maximum European Commission limits. Parsley, dill, basil, thyme, and rucola contained high concentrations of nitrate from mean levels of 2134 mg kg^–1 for parsley up to 8150 mg kg^–1 for rucola. Mean nitrate concentrations ranged from 382 to 1115 mg kg^–1 for white cabbage and Chinese cabbage, respectively. The per capita mean daily intake of nitrates related to the consumption of leafy vegetables, culinary herbs, and cucumber for the whole Estonian population was 31.3 mg day^–1, which comprised 14.2% of the acceptable daily intake (ADI).