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[目的]对直接和间接法制备酶标记三聚氰胺抗原进行比较。[方法]2种方法均用辣根过氧化物酶和过碘酸钠法,但直接法是酶与三聚氰胺偶联,而间接法是用酶标记三聚氰胺与载体蛋白OVA的结合物。用上述方法分别制备了2种酶标抗原后,从酶活性、抗原性和效价方面对这2种酶标抗原做质量对比试验。[结果]间接法制备的HRP-OVA-MEL有抗原性,且效价达到1∶2 560以上,而直接法制备的HRP-MEL却没有抗原性,且效价较低。[结论]间接法制备的三聚氰胺酶标抗原的活性比直接法好。 相似文献
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采用辣根过氧化物酶对酪蛋白进行酶促交联,并经SDS-聚丙烯酰胺凝胶电泳、光谱分析和毛细管电泳验证.利用毛细管电泳、面积归一化方法对交联酪蛋白进行分析并计算交联度;以交联度为指标、应用响应面分析法对交联条件优化,得出适宜的条件为:温度37℃、反应时间2.9 h、酶添加量为每克蛋白质4.73μkat,此条件下酪蛋白交联度达到6.9%.与酪蛋白相比,交联酪蛋白的乳化活性在蛋白质质量分数为0.01%时提高了8.7%,而其乳化稳定性在蛋白质质量分数为0.03%时提高了21.1%. 相似文献
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辣根过氧化物酶的分离制备和纯化 总被引：4，自引：0，他引：4
从辣根块茎分离和纯化了辣根过氧化物酶（ＨＲＰ）,纯化程度包括匀浆、加热、硫酸铵分级沉淀、丙酮分级沉淀和ＣＭ－２３阴离子交换层析。纯化的ＨＲＰ的Ｋ４值为２．５×１０＾５,ＲＺ值值３．００,酶的比活４０６Ｕ／ｍｇ蛋白。它的ＳＤＳ－ＰＡＧＥ显示一条带,酶亚基分子量为４０ＫＤ,等电聚焦电泳呈现三条带,酶不舔生染色呈现多条带,表明有同工酶存在。对酶的分离纯化作了一些改进,如加氨水保持ＰＨ值,加入５ｍｍｏｌ／ 相似文献
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To produce 5-reductase inhibitory compounds, resveratrol was enzymatically oxidized in a horseradish peroxidase (HRP)/H2O2 system. Ethyl acetate extract of the oxidation products showed strong 5-reductase inhibitory activity with 10%–15% organic solvents in the system, whereas without organic solvent little inhibitory activity was exhibited. The optimum pH of enzymatic oxidation for acquisition of the inhibitory activity was 4.5. The inhibitory compounds were isolated and identified as resveratroltrans-dehydrodimer and resveratrolcis-dehydrodimer by comparing with published nuclear magnetic resonance data. The two resveratrol dehydrodimers have stronger inhibitory activity than natural resveratrol dimers and trimers found inShorea species. 相似文献
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Shingo Kawai Yosuke Kobayashi Makoto Nakagawa Hideo Ohashi 《Journal of Wood Science》2006,52(4):363-366
Nonphenolic β-O-4 lignin substructure model dimer, 1,3-dihydroxy-2-(2,6-dimethoxyphenoxy)-1-(4-ethoxy-3-methoxyphenyl)propane (I) was degraded by horseradish peroxidase (HRP) in the presence of hydrogen peroxide and 1-hydroxybenztriazole (HBT). 4-Ethoxy-3-methoxybenzoic
acid (II), 1-(4-ethoxy-3-methoxyphenyl)-3-hydroxypropanone (III), 2,3-dihydroxy-1-(4-ethoxy-3-methoxyphenyl)-1-formyloxypropane (IV), 2,3-dihydroxy-1-(4-ethoxy-3-methoxyphenyl)propanone (V), 1-(4-ethoxy-3-methoxyphenyl)-1,2,3-trihydroxypropane (VI), 1-(4-ethoxy-3-methoxyphenyl)-1,2,3-trihydroxypropane-2,3-cyclic carbonate (VII), and 1-(4-ethoxy-3-methoxyphenyl)-1,2,3-trihydroxypropane-1,2-cyclic carbonate (VIII) were identified as degradation products by gas chromatography-mass spectrometry. These degradation products were qualitatively
the same as those of substrate I in the laccase/HBT system, but the yield of the products was apparently different. The products catalyzed by the HRP/H2O2/HBT system contained large amounts of the aromatic ring cleavage products IV, VII, and VIII compared with those catalyzed by the laccase/HBT system, while the amount of Cα-Cβ cleavage product II is relatively low. These results suggest that the role of HBT is not in a simple one-electron transfer between the enzymes
and substrates. 相似文献
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研究了稀土铽(Tb)对辣根生理毒性的影响。结果表明,Tb在低浓度范围内,可以诱导叶绿素的合成,高浓度时对其产生破坏作用;对于辣根过氧化物酶,酶活性先升后降;质膜透性、丙二醛含量则呈现先减后增的变化趋势。TbCl3浓度为3mg/kg时各指标达到最值;辣根中毒阈限为10mg/kg左右。通过分析,辣根过氧化物酶对Tb反应敏感,这表明过氧化物酶可能成为研究稀土毒理的生物监测手段。 相似文献
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Long term in vitro cultures of plant tissue spontaneously change at morphological, cellular and biochemical level. Horseradish tissues were cultivated in vitro for more than 16 years. Primary crown-gall tumours were induced with Agrobacterium tumefaciens on leaf explants excised from plantlets propagated in vitro. Subcultured on hormone-free MS medium, transformed tissue continued to grow in an unorganized way (TN tumour) or as teratogenous tumour (TM teratoma) forming malformed shoots which never rooted. The aim of this study was to establish the morphological and biochemical status of this long term cultures. The study was performed by verifying biochemical marker of transformation and morphogenesis, which had been investigated when the culture was established (peroxidase, PPX) as well as by introducing new markers which have not been analyzed so far in horseradish tissues (ascorbate peroxidase, APX; phenylalanine-ammonia lyase, PAL; and phenolics content). Morphological changes were observed only in teratoma tissue. Peroxidase activity and isoenzyme pattern indicated stability of horseradish tissue culture. The new biochemical markers were in a complete (PAL) and partial (APX) correlation with the PPX results and are therefore useful addition in biochemical characterisation of these tissues and can also be applied as markers of morphogenesis and tumorization. 相似文献
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辣根过氧化物酶催化氧化17β-雌二醇的研究 总被引：1，自引：0，他引：1
[目的]优化辣根过氧化物酶（HRP）催化氧化17β-雌二醇（E2）的体系。[方法]研究反应体系pH值、反应温度、H2O2浓度、反应时间、HRP浓度和E2初始浓度对E2去除效果的影响。[结果]HRP能有效催化H2O2氧化水体中的E2。HRP酶催化氧化E2的适宜条件是：pH值为6.5,反应温度25℃,H2O2与E2摩尔反应计量比（MH2O2∶ME2）为1∶2。增大HRP和E2起始反应浓度,以及适度增加H2O2用量,反应速度会明显加快,但过量的H2O2会抑制HRP的催化活性,减小E2去除率。在[E2]初始=0.07342mmol/L,[HRP]=0.07342U/ml,pH值为6.5,反应温度为25℃和MH2O2∶ME2=1∶2条件下,反应60min,E2去除率达到94.8%。[结论]该研究可为提高H2O2氧化水体中E2的去除率提供科学依据。 相似文献