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In order to optimize enzymolysis technology for production of antioxidant peptides from chickpea protein, effect of ratio of enzyme to substrate, enzymolysis temperature, and hydrolysis time on the technology in which reducibility and superoxide anion radical capturing rate were taken as response values was analyzed with response surface methodology.

为优化Alcalase蛋白酶酶解鹰嘴豆蛋白制备抗氧化肽的工艺条件,采用响应面分析法,以还原能力、超氧阴离子捕获率为响应值,研究了酶与底物的比值、酶解温度和酶解时间对制备抗氧化肽工艺的影响。

Ratio of enzyme to substrate, temperature, and hydrolysis time for the production of antioxidant peptides were analyzed with Response surface methodology in which reducibility and superoxide anion radical capturing rate was response value to build a predictive model for the enzymolysis of chickpea protein and the optimization of the enzymolysis technology parameters.

为优化Alcalase蛋白酶酶解鹰嘴豆蛋白制备抗氧化肽的工艺条件,采用响应面分析法(response surface methodology,简称RSM),以还原能力、超氧阴离子捕获率为响应值,研究了酶与底物的比值、酶解温度和酶解时间对制备抗氧化肽工艺的影响。

The results demonstrated that the measured molecular weight of phycocyanin was 37.715Kd, and consisted of two peptides which were similar with their molecular weight and amino acid sequences.4 The comformation of phycocyaninPhycocyanin high structure was investigated, as the result of circular dichroism spectrum and Fourier transformation infared spectrum shown, the secondary structure of NSPC was mainlyα-helix structure and the content ofβ-shee or coil was very small.

根据氨基酸自动分析仪测定的氨基酸组成和精确分子质量计算出NSPC分子总共有331个氨基酸残基,分别是由40个Asp、22个Thr、18个Ser、22个Glu、1个Gly、31个Ala、27个Cys、17个Val、13个Ile、22个Leu、5个Tyr、7个Phe、7个Lys、24个His、51个Arg、14个Pro、1个Met和9个Trp组成。胰蛋白水解酶分别酶解NSPC-1和NSPC-2后,分别采用MALDI-TOF-MS分析,其肽指纹谱图显示胰蛋白酶可将NSPC-1和NSPC-2水解成大小不等的多条肽段,质量数在800以上的NSPC-2有11段,即水解的肽质量分别为899.54、1454.77、1544.9、1683.87、1803.93、1811.91、1987.92、2003.90、2536.26、2915.42和3492.72。

In this paper we combined three chromatographic separation and purification technique such as affinity chromatography, ion exchanger chromatography and hydrophobic interaction chromatography to develope a new technology of stimutaneous extraction of three enzyme from pancreatin. We optimized the technology by studying the methods of purification and assured the technology as: The crude extraction from the dissolution of Pancreatin is directly absorbed on the DEAE gelose fast flow columnEquilibrating buffer is 0.01mol/L NaoAc-HoAc buffer(pH4.5; eluting buffer is 0.2~0.35mol/LNaCl in 0.01mol/LNaoAc-HoAc buffer (pH4.5), and then be eluted by two steps to acquire the peak of kallikrein.The solution which can"t be adsorbed by DEAE gelose fast flow column is adsorbed on affinity chromatographic column Equilibrating buffer is 0.01mol/LTris-HCl buffer(pH7.5, eluting buffer is 0.5mol/LNaCl in 0.01mol/Ltris-HCl buffer(pH7.5)and then be eluted by one step to acquire the peak of trypsin.The solution which can"t be adsorbed by is pretreated with 30%~80%(NH_4)_2SO_4 fractional precipitation, the deposition of the precipitation is dissolved to beabsorbed on phenyl gelose fast flow columnhydrophobic interaction chromatography condition is Equilibrating buffer is lmol/L(NH_4_2SO_4 in 0.01mol/LNaoAc-HoAc buffer(pH4.5), eluting buffer is 0~0.6mol/L(NH_4)_2SO_4 in 0.01mol/LNaoAc-HoAc buffer (pH4.5) and then be eluted by two steps to acquire the peak of chymotrypsin.

本研究考察了各种纯化方法,将离子交换层析、亲和层析和疏水层析三种分离纯化法相结合,建立了激肽释放酶、胰蛋白酶和糜蛋白酶三酶的联产工艺:胰酶用pH4.5醋酸缓冲溶液提取后,粗提液直接上DEAE-琼脂糖快胶柱吸附平衡缓冲液:0.01mol/LNaoAc-HoAc缓冲液(pH4.5,洗脱缓冲液:0.01mol/LNaoAc-HoAc缓冲液(pH4.5)含0.2~0.35mol/LNaCl分两步洗脱,收集激肽释放酶的洗脱峰;DEAE-琼脂糖快胶的未吸附液上亲和层析柱分批吸附平衡缓冲液:0.01mol/LTris-HCl缓冲液(pH7.5,洗脱液:0.5mol/LNaCl溶液,一次洗脱,收集胰蛋白酶洗脱峰;最后,亲和层析未吸附液用30%~80%硫酸铵分级盐析处理,沉淀溶解后用上苯基—琼脂糖快胶吸附平衡缓冲液:0.01mol/LNaAc-HAc缓冲液(pH4.5含1mol/L(NH_4)_2SO_4,洗脱缓冲液:0.01mol/LNaAc-HAc缓冲液(pH4.5)含0~0.6mol/L(NH_4)_2SO_4,分两步洗脱,收集糜蛋白酶的洗脱峰。

Assulta and the induction of GST activity by nicotine and capsicine in H. assulta provided partial supports for the hypothesis that diet breadth was governed by detoxification abilities. The results also indicated that two sibling species had different strategies of adaptation to toxic substances: H. assulta saves expenditure of nutrients and energy by restricting their countermeasures to a few potentially toxic compounds. H. armigera, by contrast, must be prepared to counter a wider range of potential toxins. 5. Compared with parental species, hybrids of reciprocal hybridization had significant difference in feeding preference for hot pepper.

结果显示,广食性的棉铃虫在中肠羧酸酯酶和谷胱甘肽-S-转移酶活性方面高于寡食性的烟青虫,但烟青虫谷胱甘肽-S-转移酶活性可以被寄主植物内的烟碱和辣椒素所诱导,表明解毒酶活性在一定程度上影响着植食性昆虫的食性广度;同时也暗示着二近缘种对有毒的次生物质采取不同的适应策略:烟青虫有选择地对付种类较少的植物次生物质以减轻营养和能量的消耗;棉铃虫则必须对付存在于其寄主植物内的种类繁多的有毒次生物质。

Angiotensin-converting enzyme also breaks down BRADYKININ, a powerful vasodilator and component of the KALLIKREIN-KININ SYSTEM.

血管紧张素转换酶也分解缓激肽,一种有力的血管扩张剂和激肽释放酶-激肽系统的成分。

The blood samples were collected at day 1 before and after the caprine placenta peptide were given for 10 days, and the number of the white blood cell and the ANAE+ in periperal blood ,the content of the globulin, the activity of lysozyme, the T-Aoc ,the activity of the GSH-Px and SOD, and the content of the MDA in serum were determined .

在口服羊胎盘肽的前1d和连续口服羊胎盘肽10d后的第1d各组采血样一次,测定外周血白细胞数、ANAE+淋巴细胞百分率、血清球蛋白含量、血清溶菌酶活性、血清总抗氧化能力、血清谷胱甘肽过氧化物酶活性、血清超氧化物歧化酶活性及丙二醛含量。

The former is necessary for equilibrium-controlled synthesis of peptide bonds while the latter is necessary for kinetically controlled synthesis of peptide bonds.

酰胺酶活性起水解或合成肽键的作用,因而对平衡控制的肽键合成是必需的;酯酶活性起转移酰基的作用,因而对动力学控制的肽键合成是必需的。

To understand the toxicity mechanisms of mixed heavy metals on aquatic plant,indicators of algea growth rate,content of reduced glutathione,activities of glutathione S-transferase and glutathione peroxidase of green algae, Scenedesmus quadricauda were measured to analyze the individual and joint toxic effects of lead and mercury.

为进一步了解重金属复合污染对水生植物产生毒害作用的机制,以单细胞藻类生长及细胞内还原型谷胱甘肽含量、谷胱甘肽-S-转移酶和谷胱甘肽过氧化物酶活性为指标,对铅和汞单一及联合胁迫对四尾栅藻的毒性作用进行了研究。

The effects and mechanism of GABAergic neurons, NOergic neurons, opioid peptide and cyclic adenosine monophosphate in the nucleus reticularis thalami on sleep-wakefulness cycle of rats and the effects and mechanism of the 5-HTergic nerve fibers project from the nucleus raphes dorsalis to RT on sleep-wakefulness cycle of rats were investigated with the methods of brain stereotaxic, nucleus spile, microinjection and polysomngraphy.1. The effects of GABAergic neurons in RT on sleep-wakefulness cycle of rats1.1 Microinjection of 3-mercaptopropionic acid (3-MP, a kind of glutamate decarboxylase inhibitor) into RT. On the day of microinjection, sleep only decreased a litter. On the second day, sleep marked decreased and wakefulness marked increased. On the third and fourth day, sleep and wakefulness stages resumed to normal.1.2 Microinjection of gamma-amino butyric acid (GABA 1.0μg) into RT enhanced sleep and reduced wakefulness compared with control; while microinjection of L-glutamate (L-Glu, 0.2μg) decreased sleep and increased wakefulness; microinjection of bicuculline (BIC, 1.0μg), a GABAA receptor antagonist, enhanced wakefulness and reduced sleep; microinjection of baclofen (BAC, 1.0μg), GABAB receptor agonist, had the same effects as GABA.2. The effects of NOergic neurons in RT on sleep-wakefulness cycle of rats2.1 Microinjection of L-arginine (L-Arg, 0.5μg) into RT decreased sleep compared with control, but there were on statistaical difference between L-Arg group and control; while microinjection of sodium nitroprusside (SNP, 0.2μg), a NO donor into RT, sleep marked decreased and wakefulness marked increased. Microinjection of nitric oxide synthase inhibitor, N-nitro-L-arginine (L-NNA, 2.0μg) into RT enhanced sleep and reduced wakefulness.2.2 After simultaneous microinjection of L-NNA (2.0μg) and SNP (0.2μg) into RT, SNP abolished the sleep-promoting effect of L-NNA compared with L-NNA group; after simultaneous microinjection of L-NNA (2.0μg) and L-Arg(0.5μg) into RT, we found that L-NNA could not blocked the wakefulness-promoting effect of L-Arg.3. The effects of opioid peptide in RT on sleep-wakefulness cycle of rats3.1 Microinjection of morphine sulfate (MOR, 1.0μg) into RT increased wakefulness and decreased sleep compared with control; while microinjection of naloxone hydrochloride (NAL, 1.0μg), the antagonist of opiate receptors, into RT, enhanced sleep and reduced wakefulness.3.2 After simultaneous microinjection of MOR (1.0μg) and NAL (1.0μg) into RT, the wakefulness-promoting effect of MOR and the sleep-promoting effect of NAL were not observed compared with control.4. The effects of cAMP in RT on sleep-wakefulness cycle of rats Microinjection of cAMP (1.0μg) into RT increased sleep and decreased wakefulness compared with control; microinjection of methylene blue (MB,1.0μg) into RT enhanced sleep and reduced wakefulness compared with control.5. The effects of the 5-HTergic nerve fibers project from DRN to RT on sleep-wakefulness cycle of rats5.1 When L-Glu (0.2μg) was microinjected into DRN and normal sodium (NS,1.0μg) was microinjected into bilateral RT. We found that sleep was decreased and wakefulness was increased compared with control; when L-Glu (0.2μg) was microinjected into DRN and methysergide (MS,1.0μg), a non-selective 5-HT antagonist, was microinjected into bilateral RT, We found that sleep was enhanced and wakefulness was reduced compared with L-Glu group.5.2 When p-chlorophenylalanine (PCPA, 10μg) was microinjected into DRN and NS (1.0μg) was microinjected into bilateral RT, We found that sleep was increased and wakefulness was decreased compared with control; microinjection of 5-hydroxytryptaphan (5-HTP, 1.0μg), which can convert to 5-HT by the enzyme tryptophane hydroxylase and enhance 5-HT into bilateral RT, could block the effect of microinjection of PCPA into DRN on sleep-wakefulness cycle.

本研究采用脑立体定位、核团插管、微量注射、多导睡眠描记等方法,研究丘脑网状核(nucleus reticularis thalami,RT)中γ-氨基丁酸(gamma-amino butyric acid ,GABA)能神经元、一氧化氮(nitrogen monoxidum,NO)能神经元、阿片肽类神经递质、环一磷酸腺苷(cyclic adenosine monophosphate,cAMP)及中缝背核(nucleus raphes dorsalis,DRN)至RT的5-羟色胺(5-hydroxytryptamine,5-HT)能神经纤维投射对大鼠睡眠-觉醒周期的影响及其作用机制。1 RT内GABA能神经元对大鼠睡眠-觉醒周期的影响1.1大鼠RT内微量注射GABA合成关键酶抑制剂3-巯基丙酸(3-MP,5μg),注射当天睡眠时间略有减少,第二日睡眠时间显著减少,觉醒时间明显增多,第三、四日睡眠和觉醒时间逐渐恢复至正常。1.2大鼠RT内微量注射GABA受体激动剂GABA( 1.0μg)后,与生理盐水组比较,睡眠时间增加,觉醒时间减少;而RT内微量注射L-谷氨酸(glutamic acid, L-Glu, 0.2μg)后,睡眠时间减少,觉醒时间增加;RT内微量注射GABAA受体阻断剂荷包牡丹碱(bicuculline,BIC,1.0μg)后,睡眠时间减少,觉醒时间增加;RT内微量注射GABAB受体激动剂氯苯氨丁酸(baclofen,BAC,1.0μg)后,产生了与GABA相似的促睡眠效果。2 RT内NO能神经元对大鼠睡眠-觉醒周期的影响2.1大鼠RT内微量注射NO的前体L-精氨酸(L-Arg,0.5μg)后,与生理盐水组对比,睡眠时间略有减少,但无显著性意义;而RT内微量注射NO的供体硝普钠(Sodium Nitroprusside,SNP,0.2μg)后可明显增加觉醒时间,缩短睡眠时间;微量注射一氧化氮合酶抑制剂L-硝基精氨酸(L-arginine,L-NNA,2.0μg)后,引起睡眠时间增多,觉醒时间减少。2.2大鼠RT内同时微量注射L-NNA(2.0μg)和SNP(0.2μg)后与L-NNA组比较发现SNP逆转了L-NNA的促睡眠作用;RT内同时微量注射L-NNA(2.0μg)和L-Arg(0.5μg)后,与L-NNA(2.0μg)组比较发现L-Arg可以增加觉醒而缩短睡眠,其促觉醒作用未能被NOS的抑制剂L-NNA所逆转。3 RT内阿片肽对大鼠睡眠-觉醒周期的影响3.1大鼠RT内微量注射硫酸吗啡(morphine sulfate,MOR,1.0μg)后与生理盐水组对比,睡眠时间减少而觉醒时间增加; RT内微量注射阿片肽受体拮抗剂盐酸纳洛酮(naloxone hydrochloride,NAL,1.0μg)后与生理盐水组比较,睡眠时间增加而觉醒时间减少。3.2大鼠RT内同时微量注射MOR(1.0μg)和NAL(1.0μg)后,与生理盐水组对比,原有的MOR促觉醒效果和NAL的促睡眠效果都没有表现。4 RT内环一磷酸腺苷信使对大鼠睡眠-觉醒周期的影响大鼠RT内微量注射cAMP(1.0μg)后与NS(1.0μg)组比较,睡眠时间增多而觉醒时间减少;RT内微量注射亚甲蓝(methylene blue,MB,1.0μg)后,与NS组比较,睡眠时间增多而觉醒时间减少。5中缝背核投射到丘脑网状核的5-羟色胺能神经纤维对大鼠睡眠-觉醒周期的影响5.1大鼠DRN内微量注射L-Glu(0.2μg),同时在双侧RT内微量注射NS (1.0μg)后,与对照组(DRN和双侧RT注射NS, 0.2μg)比较,睡眠时间减少,觉醒时间增多;大鼠DRN内微量注射L-Glu(0.2μg),同时在双侧RT内微量注射二甲基麦角新碱(methysergide, MS, 1.0μg )后,与对照组(DRN注射L-Glu 0.2μg,双侧RT注射NS 1.0μg)比较,睡眠时间增多,觉醒时间减少。5.2大鼠DRN内微量注射对氯苯丙氨酸(p-chlorophenylalanine,PCPA,10μg),同时在双侧RT内微量注射NS (1.0μg)后,与对照组(DRN和双侧RT注射NS, 1.0μg)比较,睡眠时间增多,觉醒时间减少;大鼠DRN内微量注射PCPA(10μg),产生睡眠增多效应后,在双侧RT内微量注射5-羟色胺酸(5-hydroxytryptaphan , 5-HTP, 1.0μg )后,与对照组(DRN注射PCPA 10μg,双侧RT注射NS 1.0μg)比较,睡眠时间减少,觉醒时间增多。

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呼气,收缩臀部肌肉;拱起身体,尽量抬起头来,右腿伸直朝向天花板(膝微屈,以避免肌肉紧张)。

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