高效液相色谱法测定17AA氨基酸注射液中N-乙酰-L-半胱氨酸和N-乙酰-L-酪氨酸

建立了一种用高效液相色谱测定 17AA氨基酸注射液中N 乙酰 L 半胱氨酸和N 乙酰 L 酪氨酸含量的方法。样品经稀释后 ,直接上机测定 ,其色谱条件为 :WatersSymmetryC18色谱柱 ,流动相为 8.5mmol/LNaAc - 5 %CH3 OH (pH =4.0 ) ,柱温36℃ ,流速 1.0ml/min ,检测波长 195nm ,N 乙酰 L 半胱氨酸在 5 .4mg/L到 5 40mg/L、N 乙酰 L 酪氨酸在 4.9mg/L到 490mg/L之间 ,标准曲线呈良好的线性关系 (r2 >0 .9999)。该方法具有良好的重现性 ,日内相对标准偏差小于 3% ,日间相对平均误差小于 7% ,样品回收率在 90 %～ 110 %之间。该方法与测定氨基酸注射液的其它高效液相色谱法相结合 ,能够对 17AA氨基酸注射液中所有氨基酸成分进行全面检测。     

N-乙酰-L-半胱氨酸减轻大鼠胃缺血/再灌注损伤的机制

为研究N-乙酰-L-半胱氨酸(N-acetylcysteine,NAC)减轻大鼠胃缺血/再灌注(gastric ischemia/reperfusion,GI/R)损伤的机制,在大鼠股静脉注射NAC(150mg/kg),夹闭大鼠腹腔动脉30min,再灌注1h制备GI/R模型。取胃后计数胃黏膜损伤指数(gastric mucosal damage index,GMDI),用原位检测(TUNEL)法观察胃黏膜细胞的凋亡,用免疫印迹法测定胃黏膜组织中p-ERK,p-JNK和NF-κB的表达,用RT-PCR法检测TNF-α,Caspase-3的mRNA表达。结果显示,NAC可以减轻GI/R损伤大鼠胃黏膜细胞的凋亡;促进大鼠I/R损伤的胃黏膜组织中p-ERK蛋白的表达,抑制p-JNK和NF-κB的蛋白表达,同时也抑制TNF-α mRNA和Caspase-3 mRNA的表达。股静脉给予辣椒素受体(vanilloid receptor subtype1,VR1)拮抗剂(capsaizepin,CPZ)400mg/kg,能逆转NAC对大鼠GI/R损伤的保护作用。以上结果提示:NAC对大鼠GI/R损伤具有减轻作用,其保护机制可能是通过上调胃黏膜组织中p-ERK,下调p-JNK和NF-κB实现的,且这种保护作用可能与VR1有关。     

S-苯基-L-半胱氨酸测定方法的研究

建立了用旋光度法和紫外分光光度法分析测定S 苯基 L 半胱氨酸的方法。研究表明 ,旋光度法适用于提纯后的产品中S 苯基 L 半胱氨酸含量的测定 ,而紫外分光光度法适用于初始产品中S 苯基 L 半胱氨酸含量的测定     

反相HPLC法快速测定S-苯基-L-半胱氨酸

建立了用反相HPLC法分析测定S-苯基-L-半胱氨酸的方法,操作简便、结果准确。方法的RSD为1.9%,加标回收率为97%~102%。     

N-乙酰-L-半胱氨酸的合成及其在医药上的应用

概述了近几十年来国内外关于Ｎ－乙酰－Ｌ－半胱氨酸的合成方法及其在医药上的应用。     

酶法转化DL-ATC合成L-半胱氨酸的酶促反应条件研究

目的:考察酶源保存方式、酶促反应时间、底物pH值、底物浓度、酶浓度、金属离子等因素对酶活力的影响。方法:以假单胞菌(Pseudomonassp.)TS1138为供试菌株,采用酸式茚三酮法测定L-半胱氨酸含量,研究了酶法转化DL-ATC合成L-半胱氨酸的酶促反应条件。结果:TS1138菌株中L-半胱氨酸脱巯基酶具有较高的活性,而且Mg2 、Mn2 、Fe2 、Zn2 、Cu2 等5种金属离子对DL-ATC水解酶酶系有不同程度的抑制,其中Cu2 对该酶系的抑制作用很大。结论:确定了TS1138菌株酶法转化DL-ATC合成L-半胱氨酸的最适酶促反应条件,为酶促反应动力学的研究奠定了基础。     

电渗析法分离提纯N-乙酰-L-半胱氨酸研究

使用我校由辐射法制备的高性能离子交换膜ＨＦ—１及ＨＦ—２,采用电渗析技术对合成所得的Ｎ－乙酰－Ｌ－半胱氨酸进行了分离提纯,脱盐率＞１５％,损失率＜１５％,为工业化应用提供了依据。     

假单胞菌酶法转化DL-ATC合成L-半胱氨酸

采用微生物酶转化法制备L-半胱氨酸具有周期短、成本低、区域和立体选择性强、反应条件容易控制、环境友好等特点,与传统的毛发水解以及化学合成工艺相比显示出明显的优越性。本文从假单胞菌产酶条件和酶学性质、DL-ATC生物转化途径、固定化细胞转化工艺、基因工程菌的研究、以及L-半胱氨酸脱巯基酶的研究等5个方面介绍了国内外关于生物转化DL-2-氨基-Δ2-噻唑啉-4-羧酸(DL-ATC)合成L-半胱氨酸的研究进展。     

比色法测定复合氨基酸注射液中的N-乙酰-L-半胱氨酸

根据Ｎ－乙酰－Ｌ－半胱氨酸与氯化钯（ＰｄＣｌ２）反应,生成一种稳定的黄色化合物,我们建立了比色测定复合结晶氨基酸注射液中的ＮＡＣ的方法。测定波长为３７５ｎｍ,检测结果重现性好,变异系数是３．９％,回收率范围在９３．１％～１００．２％之间,检测下限为０．１６３μｇ／ｍｌ。该方法比经典的电量滴定法灵敏度高、精确性好,较高压液相色谱法操作简便、费用低。     

微生物发酵法生产L-丝氨酸及L-半胱氨酸研究进展

L-丝氨酸及L-半胱氨酸在食品、医药和化妆品等行业有着广泛的应用,在植物和微生物中两者从头合成的前体物均为3-磷酸甘油酸。微生物发酵生产这两种氨基酸以其社会、经济及环境效益展现出良好的前景。针对近年来微生物发酵法生产L-丝氨酸及L-半胱氨酸的研究成果,本文综述了当前国内外学者在该领域研究的热点,即代谢途径及其调节、相关氨基酸的转运及运输、菌种及菌株改造、新菌种的开发等。最后结合当前生物技术的新发展,对今后的研究方向进行了展望。     

D-半胱氨酸盐酸盐制备研究

D-半胱氨酸盐是手性药物,主要是第三代头孢抗生素药物—头孢米诺钠的重要中间体。本文介绍了D-半胱氨酸盐酸盐的研制。采用L-半胱氨酸盐酸盐为出发原料,经消旋、沉淀异构体、结晶分离、提纯的方法,制成达到日本味之素公司1997年公布的产品质量指标的D-半胱氨酸盐酸盐。     

几种金属单质及其合金修饰电极电解合成L-半胱氨酸的研究

用几种氢过电位较高的金属Ｓｎ、Ｂｉ、ＮｉＺｎ以及Ｃｕ修饰铅电极,用于电解合成Ｌ－半胱氨酸的反应,首先筛选出性能较好的Ｓｎ、Ｎｉ、Ｚｎ电极,然后对该三种金属的合金进行研究,筛选出性能优越的（Ｎｉ－Ｓｎ）／Ｐｂ电极,电极活性大为提高,反应同期转化率明显提高。     

Cysteine proteinases of microorganisms and viruses

This review considers properties of secreted cysteine proteinases of protozoa, bacteria, and viruses and presents information on the contemporary taxonomy of cysteine proteinases. Literature data on the structure and physicochemical and enzymatic properties of these enzymes are reviewed. High interest in cysteine proteinases is explained by the discovery of these enzymes mostly in pathogenic organisms. The role of the proteinases in pathogenesis of several severe diseases of human and animals is discussed.     

高半胱氨酸膳食对小鼠生长和免疫力的影响

探讨了高半胱氨酸膳食对小鼠生长及对外来抗原刺激产生抗体的免疫力的影响。实验取小白鼠32只,随机分为高半胱氮酸组(H)和对照组(C)各16只。H组每天给予含半胱氨酸质量分数为3％的饲料,分别在1、4、8周对其腹腔注射牛血清白蛋白,在第9周处死动物,取血测量血清抗体效价。结果显示实验组生长缓慢,血清抗体效价显著高于对照组,表明高半胱氨酸膳食可提高小鼠血清抗体效价,但高含量的半胱氨酸对小鼠生长有抑制作用。     

Cysteine inhibits amyloid fibrillation of lysozyme and directs the formation of small worm‐like aggregates through non‐covalent interactions

In this article, we discuss the effects of amino acids on amyloid aggregation of lysozyme. l ‐cysteine (Cys) dramatically inhibited fibrillation of lysozyme, whereas other amino acids (including l ‐arginine) did not. In the presence of Cys, the aggregation pathway of lysozyme shifted from fibrillation to the formation of the small worm‐like aggregates with unfolding. The interaction between Cys and lysozyme was observed to be non‐covalent, suggesting that the thiophilic interaction between the thiol group on the side chain of Cys and the core sequence of lysozyme significantly contributes to the inhibition of amyloid aggregation. These findings provide a new basis for the design of a biocompatible additive to prevent amyloid fibrillation. © 2014 American Institute of Chemical Engineers Biotechnol. Prog., 30:470–478, 2014     

Role of the Liver in Regulation of Body Cysteine and Taurine Levels: A Brief Review

The first-pass metabolism of dietary sulfur amino acids by the liver and the robust upregulation of hepatic cysteine dioxygenase activity in response to an increase in dietary protein or sulfur amino acid level gives the liver a primary role in the removal of excess cysteine and in the synthesis of taurine. Hepatic taurine synthesis is largely restricted by the low availability of cysteinesulfinate as substrate for cysteinesulfinate decarboxylase, and taurine production is increased when cysteinesulfinate increases in response to an increase in the hepatic cysteine concentration and the associated increase in cysteine dioxygenase activity. The upregulation of cysteine dioxygenase in the presence of cysteine is a consequence of diminished ubiquitination of cysteine dioxygenase and a slower rate of degradation by the 26S proteasome.     

百合巯基蛋白酶抑制剂的纯化及部分性质研究

百合的鳞茎中含有一种对木瓜蛋白酶有强抑制作用的巯基蛋白酶抑制剂．百合的鳞茎经浸取加热处理,木瓜蛋白酶偶联的Ｓｅｐｈａｒｏｓｅ４Ｂ柱亲和层析和ＳｅｐｈａｄｅｘＧ－１００分子筛层析,可获得在ＰＡＧＥ和ＳＤＳ－ＰＡＧＥ均为单一蛋白带的百合巯基蛋白酶抑制剂（ＣＰＩ）．此ＣＰＩ为单链蛋白,含有０．３０７％的中性糖;Ｎ端氨基酸为Ｉｌｅ;ＳＤＳ－ＰＡＧＥ测得亚基分子量为１２０００;ＳｅｐｈａｄｅｘＧ－１００测得分子量为１２５００．百合ＣＰＩ在１００℃内和ｐＨ２～１２范围内非常稳定;对木瓜蛋白酶的抑制属竞争性抑制类型,其Ｋｉ值为１．１５×１０~（－９）ｍｏｌ／Ｌ,对木瓜蛋白酶的抑制摩尔比为８．５：１．     

Cysteine Suppresses Oxidative Stress-Induced Myofibrillar Proteolysis in Chick Myotubes

The effects of cysteine as an antioxidant nutrient on change in protein modification and myofibrillar proteolysis in chick myotubes by induction of oxidative stress by H₂O₂ treatment were investigated. Myotubes were treated for 1 h with H₂O₂ (1 mM). After this treatment, the H₂O₂ was removed and the cells were cultured in cysteine (0.1 and 1 mM) containing serum-free medium for 24 h. Protein carbonyl content as an index of protein modification and N^τ-methylhistidine release as an index of myofibrillar proteolysis were increased at 24 h after H₂O₂ treatment, and the increment was reduced by cysteine. Calpain, proteasome and cathepsin (B+L and D) activities were increased at 24 h after H₂O₂ treatment, and the increment was also reduced by cysteine. These results indicate that cysteine suppresses protein modification by oxidative stress, resulting in a decrease of protease acitivities, finally resulting in a decrease in myofibrillar proteolysis in chick myotubes.