共查询到19条相似文献,搜索用时 140 毫秒
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酶解法制备方格星虫多肽及其抗氧化作用研究 总被引:1,自引:0,他引:1
采用木瓜蛋白酶酶解制备方格星虫多肽,通过单因素实验研究了加酶量、酶解温度、酶解时间、酶解pH值和料液比等因素对羟自由基清除率的影响。通过正交试验确定了木瓜蛋白酶对方格星虫最佳的酶解工艺条件为:加酶量300 U/g,酶解温度50℃,酶解时间60 min,酶解液pH值7.0,料水比1∶3。在此条件下酶解方格星虫获得多肽的羟自由基清除率为95.42%,表明方格星虫酶解物具有较明显的抗氧化能力。由高效体积排阻色谱(HPSEC)法测得方格星虫多肽的分子量为5868。 相似文献
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以小球藻藻种为原料,通过实验室培育得到藻粉,再通过碱性蛋白酶制备小球藻抗氧化肽,以DPPH自由基清除率为指标,通过单因素和响应面设计探究加酶量、pH、酶解时间和反应温度对抗氧化肽的抗氧化性的影响,通过超滤离心对多肽进行分离,测定了各组分的DPPH自由基清除率,再通过氨基酸分析仪和凝胶渗透色谱仪测定了DPPH自由基清除率最好的多肽组分的氨基酸组成和分子量,又测定了它的热稳定性和贮藏稳定性。结果表明:小球藻蛋白制备抗氧化肽的最佳条件为:pH为7,反应温度为40℃,反应时间为30 min,加酶量为4 000 U·g-1,此时DPPH自由基的清除率是58.03%。超滤分离得到五个组分,其中分子质量为5 KD-10 KD的DPPH自由基清除率最佳,多肽中的抗氧化氨基酸含量为49.9%,它的相对分子量为474,抗氧化肽有着良好的热稳定性以及贮藏稳定性。 相似文献
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采用水酶法制备虹鳟鱼骨油,单因素分析法优化虹鳟鱼骨酶解的工艺条件,考察了料液比、pH值、酶解时间、酶解温度、加酶量5个因素对鱼油提取率的影响,采用气相色谱-质谱联用 (GC-MS) 技术对鱼骨油的脂肪酸组成和含量进行了分析鉴定.结果表明,在55℃、pH 7.5、酶解时间为3 h、料液比为1∶1、加酶量为2000 U/g的条件下,利用碱性蛋白酶提取的虹鳟鱼骨油中的油脂含量最高.GC-MS分析结果表明,虹鳟鱼骨油中主要成分是不饱和脂肪酸,含量为脂肪酸总量的80.4% (w/w),其中单不饱和脂肪酸和多不饱和脂肪酸分别约占不饱和脂肪酸的76.9%和23.1% (w/w), DHA和EPA的总量为3.4% (w/w).本研究优化了虹鳟鱼油的提取技术,对虹鳟鱼油的主要挥发性物质进行了分析鉴定,初步确定了其中对鱼油风味起主要贡献的物质,对鱼油产品的分析与鉴别具有参考价值. 相似文献
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多孔硅球固定化木瓜蛋白酶的制备和性质 总被引:11,自引:0,他引:11
用载体交联法制备了多孔硅球固定化木瓜蛋白酶。考察了固定化时间、温度、pH值、给酶量和成二醛浓度对固定化木瓜蛋白酶活力的影响。研究了固定化木瓜蛋白酶的性质,并同溶液酶进行了比较。着重考察了固定化木瓜蛋白酶的热稳定性。所制得的固定化木瓜蛋白酶最适温育温度达到80℃,对底物酪蛋白的水解活力随温度的升高而增加,在90℃达到最高值;在70℃温育12小时后酶活力仍能保持高水平。 相似文献
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S. A. Lukogorskaya I. V. Shugalei I. V. Tselinskii 《Russian Journal of General Chemistry》2002,72(2):315-318
The intensity of spontaneous peroxide fragmentation of the polypeptide chain of hemoglobin was studied in relation fo the protein concentration and pH. A chain reaction mechanism with cross chain termination was suggested. The main chain-leading radical in the process is hydroxy radical, which is responsible for peroxide damage of other biopolymers also. 相似文献
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化学修饰木瓜蛋白酶的固定化及性质研究 总被引:1,自引:0,他引:1
在底物保护和无底物保护下,用丁二酸酐对木瓜蛋白酶进行化学修饰,以三硝基苯磺酸法测定修饰酶的平均氨基修饰度,以棉布为载体,戊二醛为交联剂,对修饰前后的木瓜蛋白酶分别进行固定化.考察了温度、pH和表面活性剂SDS对化学修饰的固定化木瓜蛋白酶活力的影响,并与固定化天然木瓜蛋白酶进行了比较.研究表明,化学修饰固定化木瓜蛋白酶的最适反应温度为80℃;最适pH为9.0;在SDS浓度为20mg/mL时酶活也仍能保持在40%左右;米氏常数为187g/L.与天然的固定化酶相比,化学修饰的固定化木瓜蛋白酶的热稳定性、耐碱性和耐洗涤性得到了显著提高. 相似文献
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用不同酸酐对木瓜蛋白酶进行化学修饰, 以三硝基苯磺酸法(TNBS)测定修饰酶的平均氨基修饰度, 对修饰前后的木瓜蛋白酶分别纯化并通过UV-vis和IR对其结构进行了表征. 考察了温度、pH值和表面活性剂SDS对化学修饰的木瓜蛋白酶活力的影响, 并与天然木瓜蛋白酶进行了比较, 对天然酶和修饰酶进行了动力学研究. 结果表明, 化学修饰木瓜蛋白酶的最适反应温度为80 ℃; 最适pH值为9.0; 在SDS浓度为5 mg•mL-1时修饰酶酶活仍能保持在50%左右; 在所有酶中, 均苯四甲酸酐修饰木瓜蛋白酶的催化效率最高, 为2.442×102. 与天然木瓜蛋白酶相比, 化学修饰木瓜蛋白酶的热稳定性、耐碱性和耐洗涤性得到了显著提高. 相似文献
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The preparation,characterization,and application of silica-coated magnetic nanoparticles for papain immobilization is reported.Papain was covalently attached onto the(3-chloropropyl) trimethoxysilane-modified silica-coated magnetic nanoparticles. The enzyme-immobilized nanoparticles were characterized by Fourier transform infrared spectroscopy,X-ray powder diffraction,scanning electron microscopy,and vibrating sample magnetometry techniques.Response surface methodology combined with statistical analyses using Minitab were employed to evaluate optimum operating conditions to immobilize papain on the magnetic nanoparticles.The optimum conditions were: temperature = 27.3℃,pH of the enzyme solution = 7.1,concentration of papain = 3.3 mg/mL,and immobilization time = 10 h.Compared with the free papain,the immobilized papain displayed enhanced enzyme activity,better tolerance to variations in the medium pH and temperature,improved storage stability,and good reusability.Both the free and immobilized enzymes were effective for the clarification of pomegranate juice. 相似文献
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溶解性可调节的酶载体制备和固定化酶的研究 总被引:5,自引:0,他引:5
本文利用自由基沉淀聚合反应,合成了甲基丙烯酸-丙烯酰胺-顺丁烯二酸酐三元共聚物,测定了这些共聚物形成水不溶性的大分子氢键复合物的临界pH值.利用共聚物上的酸酐基团,直接进行了木瓜蛋白酶的固定化,得到了具有液相酶与固相酶两者优点的新型修饰酶。 相似文献
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《先进技术聚合物》2018,29(1):575-586
In this work, a new surface‐initiating system was constituted on the surfaces of cross‐linked polyvinyl alcohol (CPVA) microspheres, and on this basis, papain surface‐imprinted material was successfully prepared in aqueous solution. CPVA microspheres were modified with chlorethamin as reagent, and so a mass of primary amino group was introduced onto CPVA microspheres. Whereupon, a surface initiating system (−NH2/S2O82−) was formed at the interface between the microspheres and aqueous solution, in which papain as template protein, 4‐styrene sulfonate (SSS) as functional monomer, N,N′‐methylenebisacrylamide (MBA) as cross‐linker and (NH4)2S2O8 as initiator were all dissolved. In neutral solution, the polypeptide chains of papain as a basic protein were positively charged, and the molecules of anionic monomer SSS would spontaneously gather around papain polypeptide chain, forming complex by right of strong electrostatic interaction. The free radicals produced on CPVA microspheres initiated the monomer SSS around papain polypeptide chain and the cross‐linker MBA to produce graft/cross‐linking polymerization, and at the same time, papain macromolecules were embed in the cross‐linked networks. As a result, the graft/cross‐linking polymerizing of SSS and the molecule imprinting of papain were synchronously carried out, and papain surface‐imprinted material, MIP‐PSSS/CPVA microspheres, was obtained. The experimental results show that the papain surface‐imprinted material has excellent binding affinity and high recognition selectivity for papain. The binding capacity of MIP‐PSSS/CPVA microspheres for papain reaches 44 mg/g, and relative to another basic protein, trypsin (TRY) as contrast protein, the selectivity coefficient of MIP‐PSSS/CPVA microspheres for papain is 14.35, displaying very high recognition specificity. 相似文献
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A kind of macroporous bead carrier of copolymer containing monomer units of N-aminoethyl acrylamide and vinylalcohol was synthesized, i.e. the MR-AA carrier. Papain was immobilized on the carrier using glutaraldehyde as the couplingagent. The enzymatic activity of the immobilized papain was compared with free papain using casein as a substrate, and theeffects of glutaraldehyde concentration, pH, temperature, time and papain amount added on the activity recovery were alsoinvestigated. The results show that the MR-AA carrier contains reactive primary amine groups,hydrophilic amido links and hydroxyl groups,as well as macroporous structures based on its matrix (MR-AV matrix),furthermore,the activity recovery of papain in the immobilization could reach 48%~58%.In comparison with free papain,the resulting immobilized papain exhibits a remarkable thermostability and better reusability. 相似文献
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Cucumisin (EC 3.4.21.25) isolated from prince melon fruit is a plant serine protease. Its milk-clotting activity was compared
with plant cysteine proteases such as papain (EC 3.4.22.2) and ficain (EC 3.4.22.3). Cucumisin was more stable than papain
under the condition of pH 7.1, 37‡C for 24 h. The milk-clotting activity of cucumisin was the same to that of papain and was
half value of that of ficain. 相似文献