Comparison of Physicochemical and Structural Properties of Acid-Soluble and Pepsin-Soluble Collagens from Blacktip Reef Shark Skin

Fish collagen has been widely used in tissue engineering (TE) applications as an implant, which is generally transplanted into target tissue with stem cells for better regeneration ability. In this case, the success rate of this research depends on the fundamental components of fish collagen such as amino acid composition, structural and rheological properties. Therefore, researchers have been trying to find an innovative raw material from marine origins for tissue engineering applications. Based on this concept, collagens such as acid-soluble (ASC) and pepsin-soluble (PSC) were extracted from a new type of cartilaginous fish, the blacktip reef shark, for the first time, and were further investigated for physicochemical, protein pattern, microstructural and peptide mapping. The study results confirmed that the extracted collagens resemble the protein pattern of type-I collagen comprising the α₁, α₂, β and γ chains. The hydrophobic amino acids were dominant in both collagens with glycine and hydroxyproline as major amino acids. From the FTIR spectra, α helix (27.72 and 26.32%), β-sheet (22.24 and 23.35%), β-turn (21.34 and 22.08%), triple helix (14.11 and 14.13%) and random coil (14.59 and 14.12%) structures of ASC and PSC were confirmed, respectively. Collagens retained their triple helical and secondary structure well. Both collagens had maximum solubility at 3% NaCl and pH 4, and had absorbance maxima at 234 nm, respectively. The peptide mapping was almost similar for ASC and PSC at pH 2, generating peptides ranging from 15 to 200 kDa, with 23 kDa as a major peptide fragment. The microstructural analysis confirmed the homogenous fibrillar nature of collagens with more interconnected networks. Overall, the preset study concluded that collagen can be extracted more efficiently without disturbing the secondary structure by pepsin treatment. Therefore, the blacktip reef shark skin could serve as a potential source for collagen extraction for the pharmaceutical and biomedical applications.     

特殊医学用途婴儿配方食品中氯的国标检测方法适用性

依据GB 5009.44—2016《食品安全国家标准 食品中氯化物的测定》,对特殊医学用途婴儿配方食品中氯的检测方法适用性展开研究。结果表明：采用GB 5009.44—2016第三法（银量法）检测特殊医学用途婴儿配方食品中乳蛋白部分水解配方、乳蛋白深度水解配方、氨基酸配方得到的样品加标回收率分别为95%～108%、98%～125%、100%～106%,相对标准偏差为12.7%～32.9%;采用GB 5009.44—2016第一法（电位滴定法）得到的检测结果低于第三法检测结果。     

稀有白甲鱼含肉率及肌肉营养成分分析

试验结果表明,稀有白甲鱼含肉率为69．92％;肌肉中蛋白质、脂肪和灰分含量分别为16．58％、2．05％、0．99％;肌肉中17种常见氨基酸（除色氨酸）总含量（占肌肉干重）为77．89％。其中必需氨基酸含量为36．27％,占氨基酸总含量的46．56％。必需氨基酸与非必需氨基酸含量的比值为0．87。鲜味氨基酸含量为26．26％,占氨基酸总含量的33．70％。必需氨基酸指数为70．44。缬氨酸和异亮氨酸分别为第一、二限制性氨基酸。     

Antioxidative Effect of Chlorella Pyrenoidosa Protein Hydrolysates and Their Application in Krill Oil-in-Water Emulsions

Chlorella pyrenoidosa is an excellent source of protein, and in this research, we assessed the antioxidant and emulsifying effects of Chlorella protein hydrolysate (CPH) using neutral proteases and alkaline proteases, as well as the properties of CPH-derived krill oil-in-water (O/W) emulsions. The CPHs exhibited the ability to scavenge several kinds of free radicals, including 1,1-diphenyl-2-picrylhydrazyl (DPPH), O₂⁻, hydroxyl, and ABTS. Additionally, the CPHs (5 mg/mL) scavenged approximately 100% of the DPPH and ABTS. The CPHs showed similar emulsifying activities to Tween 20 and excellent foaming activities (max FS 74%), which helped to stabilize the krill oil-in-water emulsion. Less than 10 mg/mL CPHs was able to form fresh krill oil-in-water emulsions; moreover, the CPHs (5 mg/mL) in a krill O/W emulsion were homogenous, opaque, and stable for at least 30 days. Based on their inhibitory effects on the peroxide value (POV) and thiobarbituric acid reactive substances (TRABS), the CPHs were found to be able to inhibit lipid oxidation in both emulsifying systems and krill O/W emulsions. Thus, the CPHs could improve superoxide dismutase (SOD) activities by 5- or 10-fold and decrease the high reactive oxygen species (ROS) level caused by the addition of H₂O₂ in vitro. In conclusion, health-promoting CPHs could be applied in krill oil-in-water emulsions as both emulsifiers and antioxidants, which could help to improve the oxidative and physical stability of emulsions.     

C-端嵌合PRRSV T细胞抗原表位对PCV2 VLPs稳定性的影响

【目的】利用大肠杆菌原核表达系统表达猪圆环病毒2型Cap蛋白及在其C-端嵌合PRRSV T细胞抗原表位的Cap嵌合蛋白,对Cap蛋白形成的病毒样颗粒(VLPs)的稳定性、免疫原性以及C-端嵌合PRRSV T细胞抗原表位VLPs的热稳定性进行研究,为猪圆环病毒2型(PCV2)VLPs疫苗的高效制备及基于Cap VLPs纳米骨架展示技术嵌合疫苗的开发奠定基础。【方法】采用分子生物学方法合成PCV2b型强毒株ZJ的cap基因,在其C-端插入PRRSV T细胞抗原表位T1、T2、T3、T4、T5,构建cap-T1～cap-T5基因;将cap及cap-T1～cap-T5基因与pET28a表达载体连接,构建重组表达载体,转染大肠杆菌ClearColi^TMBL21(DE3)进行诱导表达,并对表达产物的热稳定性及免疫效果进行检测。【结果】实现了Cap蛋白及其C-端嵌合PRRSV T细胞抗原表位的Cap-T1、Cap-T2、Cap-T3、Cap-T4和Cap-T5嵌合蛋白在大肠杆菌中的可溶性表达,并成功观察到了VLPs;表达产物热稳定性分析显示,在60℃处理30 min条件下,Ca...     

硬脂酸-胱氨酸-大豆分离蛋白复合膜的机械特性和吸湿性研究

以硬脂酸作为增塑剂,胱氨酸作为交联剂制备具有一定力学性能和良好抗湿性能的大豆分离蛋白复合膜。将膜放在25℃,相对湿度为50%的干燥器中平衡两天,用质构仪测定膜的抗拉强度(TS),延伸率(E(%))。在水分活度a_w为0.10～0.90的范围内研究了复合膜在25℃的吸湿特性。吸湿速率和吸湿等温线数据分别拟合到Peleg's 方程和GAB(Guggenheim-Anderson-de Boer)模型。结果表明：大豆蛋白复合膜的TS、延伸率E(%)以及吸湿速率随着硬脂酸和胱氨酸的添加比率显著地变化。硬脂酸和胱氨酸的最佳添加比率为40∶60(w/w)(每升蛋白质溶液中加入10 g混合添加剂),此时,大豆蛋白膜的强度比原来提高2倍,并且有最佳的吸湿速率。吸湿数据和GAB 模型有很高的拟合度,拟合系数最高达0.99。     

斗鸡新城疫病毒DQ株融合蛋白基因序列分析

从越南斗鸡暴发的病例中分离出1株新城疫病毒（NDV）,命名为贵州分离株（DQ）。采用RT—PCR方法扩增分离株（DQ）的整个F基因片段,并将其克隆到pMD-18T载体中。序列分析结果表明,DQ株F基因分子长为1662bp,编码553个氨基酸。该毒株与国内2株分离株（CH2000、TW2000）之间的亲缘关系较近,其核苷酸和氨基酸序列同源性分别为95．7％-96．6％和96．6％-96．8％;但与LaSota、B1及1748E9等常见毒株的核苷酸和氨基酸同源性仅为84．0％-86．5％和87．2％-89．9％。DQ分离株在F蛋白裂解住点的氨基酸顺序为^112R—R—Q—K—R—F^117,与NDV强毒株特征相符,且具有101处的K（赖氨酸）和121处V（缬氨酸）2个特征性氨基酸,与NDV基因Ⅶ型的特性完全相符。利用MegAlign软件绘制NDV的系统发育进化树表明该分离株为基因Ⅶd亚型。     

低温锻炼对胡杨愈伤组织抗寒性、可溶性蛋白、脯氨酸含量及抗氧化酶活性的影响

胡杨(Populus euphratica)是唯一能在中国西北部半干旱地区存活和形成森林的高大乔木树种,对干旱、盐碱、低温等逆境有较强的耐受力。本研究以胡杨愈伤组织作为材料,检测了胡杨愈伤组织在低温锻炼过程中抗寒性、可溶性蛋白含量、脯氨酸含量以及SOD、CAT、POD活性的变化。结果显示低温锻炼(2℃)不仅提高了胡杨愈伤组织的抗寒性及脯氨酸、可溶性蛋白的含量,且增强了SOD、CAT等抗氧化酶的活性。     

改良背主动脉注射法对鸡胚发育与转基因表达效率的影响

背主动脉注射是生产转基因鸡的经典方法,该方法不需换壳培养,但壳内注射技术难度大,并且无法实时观察后期胚胎发育情况.本实验对背主动脉注射法进行了改进,将增强型绿色荧光蛋白(EGFP)腺相关病毒(Adeno-associated Virus,AAV)壳外注射至150枚HH 14～16期(Hamburger-Hamilton...     

可食性绿豆淀粉基复合膜的制备及其在青脆李保鲜中的应用

以绿豆淀粉和大豆分离蛋白为制膜原料,通过考察二者复合比例对可食性绿豆淀粉基复合膜性能的影响,研究可食性绿豆淀粉基复合膜的最佳复合比例,同时利用傅里叶红外光谱仪和扫描电镜对该复合膜进行结构表征,并考察其对青脆李贮藏品质的影响。结果表明：在绿豆淀粉与大豆分离蛋白复合比例为2∶1时,绿豆淀粉基复合膜表现出良好的膜性能,此时溶解度、水蒸气透过系数、抗拉强度和断裂伸长率分别为18%、1.2×10^-5g·mm·cm^-2·h^-1·Pa^-1、35 MPa和21.4%。与绿豆淀粉膜相比,该复合膜组分间的相容性较好,且表面更平滑、均匀。在常温贮藏期间,用该复合膜涂膜的青脆李硬度、可溶性固形物和可滴定酸含量等品质指标均优于未涂膜组,可作为青脆李的保鲜膜材料。