核桃多肽的制备及核桃多肽酒的研制

采用中性蛋白酶对核桃蛋白进行水解,得到最适反应条件是：酶解4.0h,底物浓度3%,加酶量6500U/g底物,温度50℃、pH7.5,水解度达到11.6%。核桃多肽酒最佳发酵工艺条件为：温度24℃,接种量0.05%,pH4.0,时间6d。发酵结束后为了掩蔽轻微的苦味,加入0.6% β-环糊精和4%蔗糖,最终产品口感良好。     

β-环糊精鹿胎盘肽微胶囊的制备

本实验以β- 环糊精为壁材,利用超声波技术制备鹿胎盘肽微胶囊。通过正交试验研究微胶囊化过程中的最佳工艺条件。结果表明：用超声波法制备鹿胎盘肽的最佳工艺条件是：鹿胎盘肽:β- 环糊精为1:20、超声功率为160W、超声时间为100min、超声温度为30℃,此工艺条件下包埋率为60.63%。     

酶解花生蛋白制备功能性多肽条件优化研究

为优化Alcalase 蛋白酶酶解花生蛋白制备功能性多肽的工艺条件,采用响应面分析法,以水解度、短肽得率为响应值,研究温度、pH 值、底物质量分数、酶底比对制备功能性多肽工艺的影响。综合考虑成本和工艺要求等问题,最终确定酶解花生蛋白制备功能性多肽的工艺条件为温度55℃、pH8.4、底物质量分数4.31%、酶底比3.39%、时间4h。该条件下水解度(DH)及三氯乙酸可溶性氮溶解指数(TCA-NSI)分别为23.40% 和74.88%,与理论值的相对误差在0.5% 以内,优化工艺稳定,DH 及TSA-NSI 较高,实验结果与模型预测值相符。     

多肽胰岛素正常转运经淋巴而非门脉途径IGMEI胰组织注射SEM观察

探讨胰岛素等多肽激素在细胞外的正常转运途径或规律,对免疫金标记外源性胰岛素（IGMEI）注射的大鼠胰组织冷冻切片,进行了二次电子和背散射电子图像（SEI／BEI）的扫描电镜（SEM）示踪性观察,SEM观察SEI显示胰腺小叶和小叶间结缔组织,血管和淋巴管,胰腺导管和胰岛的结构特点清晰可见,根据管腔内是否存在红细胞和内皮细胞的结构特点,可以区别血管或淋巴管。SEM观察BEI表明,较强的背散射电子出现在胰结缔组织间隙,淋巴管或毛细淋巴管内;然而,胰的血管或毛细血管内,无标记金颗粒的背散射电子或BEI微弱,结果提示,注射或释放入胰组织液中的胰岛素等多肽激素或分泌颗粒,其正常转运途径或规律,可能通过淋巴而非肝门静脉途径转运进入血液。     

联用抗口蹄疫病毒重组多肽及DNA疫苗诱发豚鼠产生特异免疫应答

选择一株O型口蹄病毒（FMDV）外壳蛋白VPI中21－40位及141－160位氨基酸残基两个抗原表位,组成串联结构141－160（20AA）－21－40（20AA）－141－160（20AA）,并与大肠杆菌β－半乳糖苷酶相连,在大肠杆菌中表达了此融合蛋白。同时将编码此融合蛋白的基因克隆进真核表达载体中,构建成重组表达质粒pCDM8FZ1。将pCDM8FZ1DNA与融合蛋白混合,免疫豚鼠一次,观察豚鼠产生免疫应答情况。结果显示,豚鼠可产生抗FMDV中和抗体及特异性T细胞增殖反应,部分豚鼠能抵抗FMDV的攻击,保护率为50％,证明将DNA疫苗与重组多肽疫苗结合起来作一次免疫,是可望用于预防FMDV感染的新途径,值得进一步探索。     

抗内毒素多肽对内毒素休克模型家兔的治疗作用

目的: 研究杀菌性通透性增加蛋白模拟肽(BNEP)对内毒素休克家兔重要器官功能的影响。方法: 家兔在静脉注射内毒素(LPS)500μg·kg^-1后,立即注射BNEP5mg·kg^-1,测定、分析不同时相点平均动脉压(MABP)、脉压差、血浆LPS、谷丙转氨酶(ALT)、肌酐(Cr)、肌酸磷酸激酶同工酶(CK-MB)的变化,观察8h死亡率。结果: BNEP组MABP下降速率减慢,并均在攻毒6h后恢复到攻毒前的80%以上。而LPS对照组平均动脉压、脉压差在攻毒后维持在低水平。BNEP组攻毒后各时相点血浆LPS浓度均显著低于单纯攻毒组,脉压差在2、4、6h时相点、CK-MB在4、6h时相点、Cr在6h时相点显著低于LPS组,且8h死亡率显著低于单纯攻毒组。但两组间ALT差别不显著。结论: BNEP具有较强的中和LPS能力,对内毒素休克家兔心血管系统、肾功能改善和恢复具有一定的效果。作为严重感染的辅助治疗手段可能具备较好的应用前景。     

Inhibition of hIAPP Amyloid Aggregation and Pancreatic β‐Cell Toxicity by OH‐Terminated PAMAM Dendrimer

Human islet amyloid polypeptide (hIAPP, or amylin) forms amyloid deposits in the islets of Langerhans, a phenomenon that is associated with type‐2 diabetes impacting millions of people worldwide. Accordingly, strategies against hIAPP aggregation are essential for the prevention and eventual treatment of the disease. Here, it is shown that generation‐3 OH‐terminated poly(amidoamine) dendrimer, a polymeric nanoparticle, can effectively halt the aggregation of hIAPP and shut down hIAPP toxicity in pancreatic MIN6 and NIT‐1 cells as well as in mouse islets. This finding is supported by high‐throughput dynamic light scattering experiment and thioflavin T assay, where the rapid evolution of hIAPP nucleation and elongation processes is halted by the addition of the dendrimer up to 8 h. Discrete molecular dynamics simulations further reveal that hIAPP residues bound strongly with the dendrimer near the c‐terminal portion of the peptide, where the amyloidogenic sequence (residues 22–29) locates. Furthermore, simulations of hIAPP dimerization reveal that binding with the dendrimer significantly reduces formation of interpeptide contacts and hydrogen bonds, thereby prohibiting peptide self‐association and amyloidosis. This study points to a promising nanomedicinal strategy for combating type‐2 diabetes and may have broader implications for targeting neurological disorders whose distinct hallmark is also amyloid fibrillation.     

Thixotropic Hydrogels Composed of Self-Assembled Nanofibers of Double-Hydrophobic Elastin-Like Block Polypeptides

Physically crosslinked hydrogels with thixotropic properties attract considerable attention in the biomedical research field because their self-healing nature is useful in cell encapsulation, as injectable gels, and as bioinks for three-dimensional (3D) bioprinting. Here, we report the formation of thixotropic hydrogels containing nanofibers of double-hydrophobic elastin-like polypeptides (ELPs). The hydrogels are obtained with the double-hydrophobic ELPs at 0.5 wt%, the concentration of which is an order of magnitude lower than those for previously reported ELP hydrogels. Although the kinetics of hydrogel formation is slower for the double-hydrophobic ELP with a cell-binding sequence, the storage moduli G′ of mature hydrogels are similar regardless of the presence of a cell-binding sequence. Reversible gel–sol transitions are demonstrated in step-strain rheological measurements. The degree of recovery of the storage modulus G′ after the removal of high shear stress is improved by chemical crosslinking of nanofibers when intermolecular crosslinking is successful. This work would provide deeper insight into the structure–property relationships of the self-assembling polypeptides and a better design strategy for hydrogels with desired viscoelastic properties.     

胰蛋白酶水解鲜猪皮的条件选择

为克服酸碱法水解猪皮胶原对氨基酸的破坏,拟利用胰蛋白酶水解猪皮胶原来获得更多的氨基酸.研究发现,随着胰蛋白酶含量增加,水解度增加,当pH 7～8时,胰蛋白酶对猪皮水解的水解度达到最大值;水解反应的开始反应速度较大,30分钟后反应速度降低.胰蛋白酶水解猪皮的最佳条件为:pH 7.5,水解温度48℃,酶与猪皮比40mg/g,底物浓度25%,水解时间2h.在此条件下,猪皮的水解度可达16%.     

Injectable Polypeptide‐Protein Hydrogels for Promoting Infected Wound Healing

Protein is the key composition of all tissues, which has also been widely used in tissue engineering due to its superior biocompatibility and low immunogenicity. However, natural protein usually lacks active functions such as vascularization, osteo‐induction, and neural differentiation, which limits its further applications as a functional biomaterial. Here, based on the mimetic extracellular matrix feature of bovine serum albumin, injectable polypeptide‐protein hydrogels with vascularization and antibacterial abilities are constructed successfully via coordinative cross‐linking of sulfydryl groups with silver ions (Ag+) for the regeneration of infected wound. In this protein hydrogel system, (Ag+), acting as crosslinkers, can not only provide a sterile microenvironment and a strong and robust antibacterial ability but also introduce K₂(SL)₆K₂ (KK) polypeptide, which endows the hydrogel with vascularization behavior. Furthermore, the in vivo data show that the polypeptide‐protein hydrogel has a considerable collagen deposition and vascularization abilities in the early stage of wound healing, resulting in rapid new tissue regeneration featured with newly appeared hair follicles. Altogether, this newly developed multifunctional 3D polypeptide‐protein hydrogel with vascularization, antibacterial properties, and hair follicle promotion can be a promising approach in biomedical fields such as infected wound healing.