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1.
菜心和水稻绿叶中不同等电点的乙醇酸氧化酶 总被引:3,自引:0,他引:3
The proteins with glycolate oxidase activity from B.parachinensis Bailey and rice( Oryza sativa )green leaves were prepared respectively.From the second protein peak on DEAE\|Cellulose column,two glycolate oxidases,expressed as B.parachinensis Bailey GO Ⅲ(specific activity natove 13 2 U·mg -1 ·min -1 )and rice GOⅢ(specific activity 8 8 U·mg -1 ·min -1 ),could not migrate anywhere in 4%~20% native\|PAGE under a pH8.3 buffer system.GOⅢ's p I was about pH8.3. The protein containing B.parachinensis Bailey GOⅢ showed 67±2,43±2,and 38±2 kD in SDS PAGE,band 43±2 kD was the subunit of B.parachinensis Bailey GOⅢ.From the two proteins above,another group of glycolate oxidases,expressed as B.parachinensis Beiley GOⅠ(specific activity 5 U·mg -1 ·min -1 )and rice GOⅠ(specific activity 1 2 U·mg -1 ·min -1 ),showed only one 43±2 kD band in SDS\|PAGE,and was purified on the Sepharose\|6B column which migrated towards anode in the same native\|PAGE showing the M r about 420 kD,or 460 kD and 260 kD respectively.GOⅠ's p I was smaller than pH8.3.Antibody against B.parachinensis Bailey GOⅠ was prepared and its efficacy was about 1/1600 in ELISA.By native\|PAGE,Western blot and rocket immunoelectrophoresis,the third group of glycolate oxidases,expressed as B.parachinensis Bailey GOⅡ and rice G0Ⅱ,were confirmed in crude protein of green leaves and migrated towards cathode under the same native\|PAGE,so GOⅡ's p I was higher than pH8.3.The M r of B.parachinensis Bailey GOⅡ was about 669 kD determined by native\|PAGE Western blot.Rice GOⅠ,rice GOⅢ and rice GOⅡ showed different quantitation under different physiological conditions.Rice GOⅡ could be induced by glycolate. 相似文献
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菠菜乙醇酸氧化酶同工酶GO Ⅰ的纯化和特性 总被引:1,自引:0,他引:1
从菠菜绿叶中获得SDS-PAGE为40000±2000M 相似文献
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菠菜中不同等电点的乙醇酸氧化酶全酶分子量的测定 总被引:5,自引:0,他引:5
乙醇酸氧化酶(EC1.1.3.15,简称GO)被认为是一个由相同亚基和黄素腺瞟吟单核着酸(FMN)组成的寡聚酶“-‘’,它在不同的生理状态下具有不同的聚合态’‘,’]。所以,GO全酶Mr差异很大,甚至有时不同或同一植物有多种并存:如豌豆为100000”’;菠菜为270000、140000和70000[”‘j;黄瓜为180000和700000[‘j;南瓜为280000~320000[sj;/J’麦、大麦、菠菜、豌豆、烟草等C3植物为160000~180000;而C4植物玉米、甘蔗为290000~310000‘’‘。但GO由相同亚基所组成的观点却难于解释其等电点(PI)也存在较大的差异:如… 相似文献
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采用聚丙烯酰胺凝胶电泳分析了吉富罗非鱼肌肉、肠、肝、胰、胃不同组织中酯酶(EST)、乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)、蛋白酶同工酶组成。结果表明:吉富罗非鱼的这4种同工酶均有明显的组织特异性。EST在肌肉组织中表达量较低,EST-1在肠中表达较高,胰、肝组织中均由6种EST同工酶谱带组成;LDH-5在肌肉组织中含量较高;SOD在肝、肌肉组织中表达较高;肠、肝蛋白酶活性较高,由5种同工酶谱带组成,胰蛋白酶含有一种同工酶。该研究为杂交鱼类的酶工程、生化遗传研究提供理论基础,进一步促进吉富罗非鱼这一优良养殖品种的开发和利用。 相似文献
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菠菜乙醇酸氧化酶同工酶的亚基组成 总被引:3,自引:1,他引:2
我们首次报告 ,菠菜有GOⅠ (pI≈ 7.4 )、GOⅡ(pI≈ 9.4 )和GOⅢ (pI≈ 8.3 ) 3种乙醇酸氧化酶(GO)同工酶 ;GOⅠ只含 4 0± 2kD一种亚基 ,而GOⅡ和GOⅢ的亚基组成未被研究 ;3种同工酶之间均有免疫同源性[1-4 ] .水稻也存在 3种GO同工酶 ,其中GOⅡ (pI >8.3 )能被乙醇酸所诱导 用柱层析法纯化可获得经SDS PAGE后为 4 3± 2kD单带的水稻GOⅠ[5~ 7] .以上初步解释了前人报告GO电荷不均一的原因[5] .最近从菠菜分离得到含GOⅡ的蛋白和含GOⅢ的蛋白 ,其SDS PAGE分别为 67± 2kD和 4 0±… 相似文献
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菠菜叶片中乙醇酸氧化酶3种同工酶的生化特性 总被引:4,自引:1,他引:3
By DEAE cellulose and Sepharose 6B chromatography, the proteins containing glycolate oxidase isozymes GOⅡ and GOⅢ were extracted from spinach green leaves. The protein containing GOⅡ showed two bands of 67±2 kD and 40±2 kD in SDS PAGE whose specific activity of glycolate oxidase was 33.4 U·mg -1 ·min -1 .It migrated towards cathode in Native PAGE in pH 8.3 buffer system. pI of GOⅡwas about 9.4 detected by IEF. The protein containing GOⅢ showed three bands of 67±2 kD, 40±2 kD and 38±2 kD in SDS PAGE whose specific activity of glycolate oxidase 14.4 U·mg -1 ·min -1 and could not migrate anywhere in the same Native PAGE. pI of GOⅢ was about 8.3 detected by IEF. The 40±2 kD might be the subunits of GOⅡ and GOⅢ. Antibodies of the protein containing GOⅡ and GOⅢ were prepared respectively. GOⅡ was very unstable and could change into GOⅢ artifact; GOⅢ was also unstable and could change into GOⅠartifact whose Mr ≈470 kD and pI ≈7.4 . This GOⅠ(specific activity: 9.8 U·mg -1 ·min -1 ), showing one 40±2 kD band in SDS PAGE, could be purified on another Sepharose 6B chromatography. The specific activity of GOⅡ decreased rapidly to about half of its original value and then was relatively stable when stored in 50% glycerol at -20℃. The results above explained why GOⅡ was extracted difficultly, and GOⅢ were easily confused with GOⅠ and GOⅡ. 相似文献
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酶法制备海洋活性肽及其功能活性研究进展 总被引:1,自引:0,他引:1
海洋生物活性肽(Marine biological active peptide)是从海洋生物中提取的具有优化机体代谢环境、有益于机体健康的一类多肽。酶法制备海洋生物活性肽是目前最常用的制备方法,是通过适当的蛋白酶水解海洋生物蛋白来制备生物活性肽的一种方法。海洋生物活性肽在降血压、抗氧化、抗凝血及抗菌等方面效果显著,对治疗和预防疾病具有巨大潜力。介绍海洋生物活性肽在酶解制备及其生物学功能方面国内外研究进展,为进一步开展海洋活性多肽研究提供参考。 相似文献
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海洋无脊椎动物抗菌肽研究进展及其在食品保鲜中的应用 总被引:1,自引:0,他引:1
海洋无脊椎动物抗菌肽抑菌广谱,稳定性高,且对生物体本身无害,其应用日益引起大量研究者的关注。综述了抗菌肽的几种类型、抑菌机理,介绍了海洋无脊椎动物抗菌肽研究进展、存在的问题并分析其在食品保鲜中的应用前景。 相似文献