基因功能注释——后基因组时代面临的挑战

在已经解序的、数以百计的生物基因组中,存在大量编码未知功能蛋白的基因序列。同时,众多已知功能的酶蛋白在解序的基因组中找不到对应的基因。确定未知功能基因的功能和寻找孤儿酶对应的基因是后基因组时代面临的极具挑战性的科学任务。本文综合讨论了目前基因组中基因功能注释存在的问题及解决这些问题的策略与方法。     

淀粉水解酶在食品工业中的应用

淀粉水解酶是指一类能催化水解淀粉酶的总称,不同使用用途及生产目的使用的淀粉水解酶的类别也不相同,本文分别介绍了淀粉水解酶中的α-淀粉酶,β-淀粉酶,异淀粉酶,葡萄糖淀粉酶的作用机理、酶性质以及在食品工业中的应用情况,为生产中淀粉水解酶的应用提供一些参考。     

蒙药材悬钩子木中黄酮类化合物含量测定及体外抗氧化性研究

采用分光光度法测定了悬钩子木中黄酮类化合物的含量,通过Fenton反应测定了悬钩子木黄酮类化合物提取液清除羟基自由基的能力.结果表明,悬钩子木中黄酮类化合物平均含量为23.66mg/g,样品回收率为90%～94%.悬钩子木黄酮类化合物提取液对羟基自由基有较强的清除作用,其清除作用随黄酮类化合物含量的增高而增加.研究结果可为悬钩子木药效功能提供科学依据.     

Synergistic defensive mechanism of phytochelatins and antioxidative enzymes in Brassica chinensis L. against Cd stress

Brassica chinensis L. was chosen and exposed to different concentrations of Cd exposure to evaluate its Cd-accumulating capacity and its potential cellular defensive mechanisms. Cd accumulation in the shoots and roots of B. chinensis was up to 1348.3±461.8 and 3761.0±795.0 mg per killogram of dry weight, respectively, under 200 μmol/L of Cd exposure. Increasing Cd accumulation in the plant was accompanied by rapid accumulation of phytochelatins (PCs), and the sequestration of Cd by PCs provided a primary cellular mechanism for Cd detoxification and tolerance of B. chinensis. Furthermore, malondialdehyde formation, hydrogen peroxide content and antioxidative enzyme activities such as superoxide dismutase, catalase, guaiacol peroxidase and ascorbate peroxidase were observed in the shoots of Cd-stressed B. chinensis. Increasing enzyme activities in response to concentrations of 5 to 50 μmol/L Cd showed an efficient defense against oxidative stress, suggesting that the antioxidative system was a secondary defensive mechanism. These resulted in reduced free Cd damage and enhanced Cd accumulation and tolerance. Glutathione plays a pivotal role in these two detoxification pathways. In general, these results suggested that PCs and the antioxidative system are synergistic in combatting Cd-induced oxidative stress and that they play important roles in Cd detoxification of B. chinensis, and also give a deep understanding of the natural defensive mechanisms in plants under heavy metal stress.     

Tetracyclines inhibit parathyroid hormone-induced bone resorption in organ culture

Summary Several tetracyclines (minocycline, doxycycline, tetracycline), in levels approximating physiologic concentrations, were found to inhibit parathyroid hormone-induced bone resorption in organ culture; the specificity of this effect was demonstrated by comparison with other (non-tetracycline) types of antibiotics. The ability of tetracyclines to inhibit bone resorption is consistent with the recent proposal by Golub et al². that these antibiotics can inhibit mammalian collagenolytic enzymes by a mechanism unrelated to the drug's antibacterial efficacy, a property which could be therapeutically useful in diseases characterized by excessive collagen breakdown.     

温度和pH对驼背鲈消化酶活力的影响

采用酶学分析方法研究了温度和pH对驼背鲈3种主要消化酶(蛋白酶、脂肪酶、淀粉酶)活力的影响。结果表明:在设定的温度和pH范围内,驼背鲈消化酶活力均随着温度和pH的升高呈先升后降的趋势。其中,胃蛋白酶的最适温度为40~45℃,肠、幽门盲囊、肝蛋白酶的最适温度均为40℃;胃脂肪酶的最适温度为35℃,幽门盲囊的最适温度为40~45℃,其他部位均为40℃;胃淀粉酶的最适温度为40℃,其他部位均为35℃。胃蛋白酶的最适pH为3.2,肠、幽门盲囊、肝的最适pH均为8.2;胃、肝脂肪酶的最适pH分别为7.2、8.2,其他部位均为6.2;胃淀粉酶的最适pH为7.2~8.2,其他器官的最适pH均为7.2。各部位在最适温度和pH下,脂肪酶酶活力顺序为肠胃肝幽门盲囊,淀粉酶酶活力顺序为肠幽门盲囊胃肝;蛋白酶在最适温度和pH下的酶活力顺序分别为肠胃肝幽门盲囊、肠胃幽门盲囊肝。     

锌对糖尿病大鼠抗氧化指标的影响

观察Zn^＋对四氧嘧啶诱发的糖尿病大鼠血液流变学的影响。按200mg／kg剂量腹腔内一次性注射四氧嘧啶（Alloxan）,建立大鼠糖尿病模型。应用Zn和二甲双胍进行为期4周的灌胃治疗,每周眼眶采血1次,对大鼠的红细胞超氧化物歧化酶（SOD）和血浆过氧化脂质产物（MDA）进行检测,4周后处死动物取肝脏检测肝脏SOD活性。结果表明10mg／kg·dZnSO4组、100mg／kg·d二甲双胍加5mg／kg·dZnSO4组、100mg／kg·d二甲双胍加10me,／kg·dZnSO4组可使糖尿病大鼠显著提高红细胞、肝脏SOD酶活性和降低血浆MDA含量（P〈0．05或P〈0．01）。Zn对实验性糖尿病大鼠提高SOD、减少MDA含量有明显作用,结合二甲双胍治疗效果更佳。     

人USP14蛋白在大肠杆菌中表达纯化的初步研究

采用PCR技术从人胎脑cDNA文库中克隆了usp14(ubiquitin-specific peptidase 14)基因编码区全长序列。序列分析表明人usp14基因含有peptidase蛋白酶C19A保守区,将usp14基因与pET-28b(+)载体相连,构建表达载体pET28b/usp14;把该表达载体转入大肠杆菌Rosetta(DE3)pLyS,以Isopropyl-D-thiogalactopyranoside(IPTG)诱导25°C 6小时,USP14蛋白获得高效表达。对表达产物进行Western blotting检测,并用Ni-NTA亲和层析技术获得了纯化的人USP14蛋白,表达的目的蛋白大小约为56kDa。     

酶解鲭鱼罐头蒸煮液制备生物活性肽

以鲭鱼罐头蒸煮液为研究对象,从风味蛋白酶、中性蛋白酶和木瓜蛋白酶中筛得最佳酶为风味蛋白酶,通过单因素实验和正交试验确定风味蛋白酶的最佳酶解工艺为:5 g底物(即5 m L蒸煮液)、加酶量28.6 mg/m L、时间5 h、p H值6.0、温度50℃.按最佳工艺条件酶解,并经小分子电泳分析,证明蒸煮液主要蛋白被风味蛋白酶高效酶解;所得生物活性肽酶解液具有较好的抗氧化活性,其DPPH自由基清除活性的IC50值为31.35 mg/m L;羟自由基清除活性的IC50值为9.807 mg/m L;还原力的IC50值为7.019 mg/m L.