Quinoa (Chenopodium quinoa Willd.) protein hydrolysates with in vitro dipeptidyl peptidase IV (DPP-IV) inhibitory and antioxidant properties

The potential of quinoa to act as a source of dipeptidyl peptidase IV (DPP-IV) inhibitory and antioxidant peptides was studied. A quinoa protein isolate (QPI) with a purity of 40.73 ± 0.90% was prepared. The QPI was hydrolysed at 50 °C for 3 h with two enzyme preparations: papain (P) and a microbial papain-like enzyme (PL) to yield quinoa protein hydrolysates (QPHs). The hydrolysates were evaluated for their DPP-IV inhibitory and oxygen radical absorbance capacity (ORAC) activities. Protein hydrolysis was observed in the QPI control, possibly due to the activity of quinoa endogenous proteinases. The QPI control had significantly higher DPP-IV half maximal inhibitory concentrations (IC₅₀) and lower ORAC values than QPH-P and QPH-PL (P < 0.05). Both QPH-P and QPH-PL had similar DPP-IV IC₅₀ and ORAC values. QPH-P had a DPP-IV IC₅₀ value of 0.88 ± 0.05 mg mL⁻¹ and an ORAC activity of 501.60 ± 77.34 μmol Trolox equivalent (T.E.) g⁻¹. To our understanding, this is the first study demonstrating the in vitro DPP-IV inhibitory properties of quinoa protein hydrolysates. QPHs may have potential as functional ingredients with serum glucose lowering properties.     

克氏原螯虾丝氨酸蛋白酶类似物部分基因的克隆与鉴定

设计简并引物,PCR扩增了克氏原螯虾(Procam barus clarkii)丝氨酸蛋白酶类似物基因的部分序列,通过与GenBank登录的序列进行比对,结果显示该序列编码的氨基酸与其他物种的丝氨酸蛋白酶类似物氨基酸序列有较高的同源性。SMART软件分析发现其催化三联体结构中的丝氨酸被甘氨酸取代,不具备蛋白酶水解活性。     

棉铃虫四个丝氨酸蛋白酶抑制剂基因的克隆与表达分析

为明确棉铃虫Helicoverpa armigera丝氨酸蛋白酶抑制剂（serine protease inhibitor,serpin）的种类及其表达特性,利用PCR技术克隆棉铃虫的serpin基因,使用生物信息学软件预测其结构并进行系统进化分析,采用实时荧光定量PCR（real time quantitative PCR,RT-qPCR）技术比较serpin基因在棉铃虫不同发育阶段和组织中的表达量及取食Cry1Ac后其表达量的变化。结果表明,共获得serpin-a、serpin-b、serpin-c、serpin-e四个棉铃虫serpin基因,全长为1 119~1 254 bp,编码373~418个氨基酸,均包含一段具有反应中心环的保守结构域,且与斜纹夜蛾Spodoptera litura、草地贪夜蛾S.frugiperda等鳞翅目昆虫serpin的同源性较高。serpin-a和serpin-e在棉铃虫4龄幼虫期的表达量最高,serpin-b和serpin-c分别在成虫期和蛹期表达量最高。serpin-a在中肠和围食膜中表达量最高,serpin-b在头、中肠和表皮中表达量最高,serpin-c在头部表达量最高,serpin-e在中肠和血淋巴中的表达量显著高于其他组织。棉铃虫取食低浓度Cry1Ac后,中肠的serpin-b和serpin-e的表达量显著增加。推测不同serpin基因在棉铃虫不同发育时期和组织中可能发挥不同的作用,其中serpin-b和serpin-e可能参与棉铃虫对Cry1Ac的解毒过程。     

黄鳝溶血毒素性质的初步研究

通过磷酸缓冲液提取、Sephadex G-50凝胶层析和DEAE Fast Flow离子交换等分离纯化技术,从黄鳝(Monopterus albus)肌肉中分离到具有溶血活性的MAM-Ⅰ-2毒素,并对其性质进行了初步研究。结果表明:MAM-Ⅰ-2能使鸡血发生溶血现象,具有丝氨酸蛋白酶抑制剂和抗凝血活性,对嗜水气单胞菌也有较强的抑制作用。经SDS-PAGE凝胶电泳测定,该毒素蛋白相对分子质量约为43 000。对MAM-Ⅰ-2的前体物质MAM-Ⅰ的研究表明:MAM-Ⅰ除具有MAM-Ⅰ-2的特性外,还对大肠杆菌、枯草芽孢杆菌等细菌以及白色念珠菌有抑制作用;不同来源的红细胞对MAM-Ⅰ毒素的敏感性不同,其中牛红细胞反应最为敏感,其次是兔、鼠和鸡的红细胞。     

Activity of gut proteinases from Cry1Ab-selected colonies of the European corn borer, Ostrinia nubilalis (Lepidoptera: Crambidae)

Susceptibility to the Cry1Ab protoxin and toxin from Bacillus thuringiensis (Berliner) and activity of gut proteinases were assessed in both susceptible and Cry1Ab-selected colonies of European corn borer, Ostrinia nubilalis (Hubner). Resistance in two different selected colonies was at least 6- and 15-fold for the Cry1Ab protoxin and 108- and 484-fold for the Cry1Ab toxin. Activities of trypsin-like, chymotrypsin-like and elastase-like proteinases were variable among the colonies tested and not indicative of a major contribution to Cry1Ab resistance. Activation of the 130-kDa Cry1Ab protoxin occurred rapidly in all colonies, with no apparent differences among colonies. In addition, there were no apparent changes in activated Cry1Ab processing, indicating that proteolytic degradation was not associated with resistance. These results suggest that mechanisms other than proteolytic activation of protoxin and toxin degradation, such as target site modification may be involved in the resistance to B thuringiensis Cry1Ab in these O nubilalis colonies.     

姜黄素对糖尿病大鼠血糖的影响

以1次性腹腔注射链脲佐菌素(STZ,65 mg·kg~(-1))诱导制备糖尿病大鼠模型,采用随机分组(正常组、糖尿病组、姜黄素组),每组10只,给予相应的药物治疗8周。结果表明:与糖尿病组比较,姜黄素具有显著降低血糖的作用,增加肝脏肝糖原含量及Akt的磷酸化水平的表达。同时,HE染色观察结果也显示,姜黄素组的肝脏损伤较糖尿病组有所减轻。结果表明,姜黄素具有明显的纠正糖代谢紊乱的作用,其作用机理与其提高机体抗氧化活性,激活Akt信号通路,改善肝脏病理损伤相关联。     

Production of gluten-free wheat starch by peptidase treatment

The potential of peptidase-containing bran extracts from germinated cereals (wheat, emmer, barley) and a peptidase preparation from Aspergillus niger (AN-PEP) to degrade gluten in wheat starch below the threshold for gluten-free foods of 20 mg/kg was compared. The gluten-specific peptidase activity of the peptidases was determined by using gliadin as a protein-based substrate as well as the two celiac-active peptides PQPQLPYPQPQLPY (α-gliadin) and SQQQFPQPQQPFPQQP (γ-hordein). The peptidase activity of AN-PEP exceeded the activities of bran from germinated cereals by a factor up to 690,000. Three wheat starches with initial gluten contents of 110, 1679, and 2070 mg/kg, respectively, were incubated with bran extracts and AN-PEP, lyophilized, and residual gluten was quantitated by a competitive ELISA. Unlike peptidases from bran extracts, AN-PEP was capable of degrading gluten below 20 mg/kg in all starches. The absence of gluten in AN-PEP-treated starches was confirmed by liquid-chromatography-mass spectrometry. The properties of gluten-free starches were comparable to the native starches with the exception of a reduced viscosity after AN-PEP treatment. This problem could be overcome by using higher enzyme concentrations and shorter incubation times or by optimizing AN-PEP production for lower residual α-amylase activity.     

乳清多肽营养饮料的研究

本文以肽酶在pH8．0、温度45℃，[E/S]3％的最适条件下水解乳清蛋白，经脱苦处理后，生产乳清多肽营养饮料；当白砂糖8％、酸0．15％、木糖醇4％时，乳清多肽饮料品质最好，含有大量人体易吸收的小分子肽，且稳定性良好。     

顶复器门原虫丝氨酸蛋白酶抑制剂研究进展

寄生虫可以利用蛋白酶抑制剂在宿主中生存,蛋白酶在宿主-寄生虫关系中处于关键地位.而寄生虫源的蛋白酶抑制剂的研究并未引起应有的重视.论文介绍了顶复器门原虫主要的内源的蛋白酶抑制剂丝氨酸蛋白酶抑制剂的研究概况,以期为顶复器门原虫的入侵机制及防治等研究提供新的思路.