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ABSTRACT: Effects of microbial transglutaminase (MTGase), recombinant cystatin, or their combination on the gel properties of mackerel surimi were investigated. The addition of MTGase caused the cross-linking of myosin heavy chain (MHC) and substantially increased the gel strength (from 536.6 to 2012.4 g × cm), while the recombinant cystatin could effectively prevent the MHC degradation and gel softening during the production of mackerel surimi-based products. Combined use of MTGase and recombinant cystatin revealed synergistic effectiveness on improving the quality of mackerel surimi (increased from 435 to 2438 g × cm, set at 45 °C for 20 min).  相似文献
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ABSTRACT: Effects of nicotinamide dinucleotide phosphate (NADPH)-sulfite reductase, recombinant cystatin, and microbial transglutaminase (MTGase) or their combination on hairtail surimi quality were investigated. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that adding NADPH-sulfite reductase could effectively recover native myosin. Apparent cross-linking of hairtail myosin during the setting process was observed with added MTGase, while myosin degradation on the heating process was substantially retarded with recombinant chicken cystatin. The combined use of NADPH-sulfite reductase, recombinant chicken cystatin, and MTGase exhibited a greater effect on improving hairtail surimi quality.  相似文献
3.
Li DK  Lin H  Kim SM 《Journal of food science》2007,72(5):C294-C299
ABSTRACT:  Recombinant chum salmon cystatin (RC) expressed in Saccharomyces cerevisiae was purified by His-select nickel affinity chromatography. The specific inhibitory activities of RC against papain and cathepsin L were 7.45 and 10.24 U/mg, respectively. RC was stable over pH 5.0 to 7.0 and at temperature below 65 °C. RC was used to prevent the gel weakening of Alaska pollock surimi. RC at 100 μg/g showed the highest inhibitory activity against the autolysis of surimi based on the analysis of TCA-soluble peptides. As the concentration of RC increased, both the breaking force and deformation of modori gel greatly increased ( P < 0.05). The addition of RC resulted in less expressible drip, which coincided with the increase of whiteness. More myosin heavy chain (MHC) was retained as the addition of RC increased. Therefore, RC could prevent the degradation of proteins in Alaska pollock surimi and was better than egg white (EW). Thus, RC could be applied to Alaska pollock surimi to prevent gel weakening and RC at 100 μg/g was the optimal concentration.  相似文献
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ABSTRACT: A DNA-encoding thioredoxin-carp ovarian cystatin (trx-cystatin) was ligated into pET-23a(+) and transformed into Escherichia coli AD494(DE3). High level of soluble recombinant trx-cystatin, expressed in E. coli was purified by 5 min of heating at 70 °C, Q-Sepahrose HP, and Sephacryl S-100 HR chromatographs. Its molecular mass was 28 kDa. It could be cleaved into a recombinant thioredoxin (16 kDa) and a mature carp ovarian cystatin (12 kDa) by enterokinase. The 12-kDa mature carp ovarian cystatin was further purified by FPLC Superdex 75 chromatography. Both recombinant trx-fused and carp ovarian cystatins were thermostable proteins and exhibited papain-like protease inhibition activity comparable to the wild-type cystatin.  相似文献
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本实验主要对鲢鱼半胱氨酸蛋白酶抑制因子Cystatin进行原核表达和纯化鉴定,并研究重组Cystatin对铜绿假单胞菌的抑菌效果。首先用异丙基-β-D-硫代半乳糖苷(isopropyl-β-D-thiogalactopyranoside,IPTG)诱导转入p ET-30-Cystatin的E.coli BL21(DE3)表达重组Cystatin蛋白,经Ni2+-NTA镍离子亲和层析对其进行纯化,该重组蛋白在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳上显示单一条带,分子质量约20.6 k D,在TSK-GEL G2000SW凝胶过滤高效液相色谱上呈单一活性峰,纯度为94.27%,经标准曲线计算其分子质量为20.9 k D;其次,利用抑制剂滴定曲线法,以偶氮酪蛋白为底物,确定重组Cystatin对木瓜蛋白酶(45.375μmol)的一个抑制活性单位为0.718μg;最后通过滤纸片扩散法鉴定鲢鱼重组Cystatin对铜绿假单胞菌的抑菌效果,结果表明Cystatin的添加量为20、60、120、200 U/片时,抑菌圈直径分别为8、9、13、26 mm。鲢鱼重组Cystatin对铜绿假单胞菌的抑菌效果呈剂量依赖关系。  相似文献
6.
本文综述了鱼类半胱氨酸蛋白酶抑制因子Cystatins超家族的分类、结构及抑制机制,进一步根据其特性,提出Cystatin有望成为鱼肉品质相关候选因子,并且阐述了其在抑制鱼糜凝胶软化及水产品抑菌、保鲜等方面的应用前景.  相似文献
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BACKGROUND: Higher plants possess several mechanisms of defense against plant pathogens. Proteins actively synthesized in response to those stresses are called defense‐related proteins which, among others, include certain protease inhibitors. It is of particular relevance to investigate plant natural defense mechanisms for pathogen control which include cystatins—specific inhibitors of cysteine proteases. RESULTS: In this study, a cysteine proteinase inhibitor (CPI), 11 kDa in size, was purified from green kiwifruit to homogeneity. Immuno‐tissue print results indicated that CPI is most abundant in the outer layer of pericarp, near the peel, and the inner most part of the pulp—sites where it could act as a natural barrier against pathogens entering the fruit. The purified protein (15 µmol L−1) showed antifungal activity against two phytopathogenic fungi (Alternaria radicina and Botrytis cinerea) by inhibiting fungal spore germination. In vivo, CPI (10 µmol L−1) was able to prevent artificial infection of apple and carrot with spore suspension of B. cinerea and A. radicina, respectively. It also exerted activity on both intracellular and fermentation fluid proteinases. CONCLUSION: Identification and characterization of plant defense molecules is the first step towards creation of improved methods for pathogen control based on naturally occurring molecules. Copyright © 2012 Society of Chemical Industry  相似文献
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