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.     

牛磺酸对鲫鱼脂肪消化吸收的影响

将0.20%、0.40%、0.60%、0.80%的牛磺酸添加到基础饲料中,以基础饲料为对照,饲喂鲫鱼(45~52 g)15 d后,测定脂肪消化吸收率。试验结果表明:牛磺酸可极显著地提高鲫鱼对脂肪的消化吸收率(P<0.01);其中,0.40%组效果最佳,比对照组提高10.19%。脂肪消化率的提高,是牛磺酸促进鱼体生长速度加快的主要原因之一。     

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。     

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。     

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。     

1株黄曲霉颉颃菌的鉴定及其抑菌活性成分分析

【目的】筛选黄曲霉颉颃菌,探究其最佳培养条件及有效抗菌成分,为实际生产中应用生物防治方法抑制黄曲霉污染提供新的理论依据。【方法】通过平板对峙法及分子生物学鉴定方法对待测菌株进行筛选鉴定。以抑菌率作为衡量标准,判断所得黄曲霉颉颃菌的最佳培养温度、培养时间、接种量、转速、pH及装液量。通过硫酸铵沉淀法从颉颃菌无菌发酵上清液中提取蛋白,观测粗提物对黄曲霉菌丝生长的影响。通过离子交换色谱和凝胶过滤色谱进一步纯化粗提蛋白,并进行质谱测序及序列对比分析,筛选鉴定颉颃菌产生的有效抑菌成分。【结果】筛选到1株解淀粉芽孢杆菌B10(菌种保藏号CCTCCNO:M2018353)。最佳培养条件为温度40℃,培养时间36 h,接种量3%,转速120 r/min, pH 6.0,装液量30 mL,最大抑菌率可达46.56%。其粗提蛋白能有效抑制黄曲霉生长,破坏菌丝正常形态,并从中筛选出几丁质结合蛋白、壳聚糖酶、葡聚糖酶等8种可能的有效抑菌成分。【结论】本试验分离鉴定出1株抑制黄曲霉正常生长发育的解淀粉芽孢杆菌B10,其产生的多种蛋白成分对黄曲霉有显著抑菌活性。     

烟粉虱模式识别受体βGRPs的先天免疫应答

为探索烟粉虱的β-1,3-葡聚糖识别蛋白(BtβGRP)在烟粉虱先天免疫过程中的重要作用,对烟粉虱中鉴定的3个BtβGRPs基因编码的蛋白质进行结构分析,并与其它物种的23个同源蛋白进行系统进化分析,同时通过实时荧光定量PCR(quantitative real-time PCR,q RT-PCR)检测分析球孢白僵菌侵染烟粉虱不同虫态不同时间点后BtβGRPs的时间表达模式。结果显示:经鉴定3个BtβGRPs蛋白序列均含有βGRP家族特有的保守性结构域,即糖苷水解酶活性结构域;且3个BtβGRPs独自聚在进化树的一支上,可能参与烟粉虱特有的级联路径;3个BtβGRPs基因在不同虫态和不同诱导时间点与对照相比,除BtβGRP3在卵期未表达外,均有上调表达趋势,但诱导程度不同;成虫诱导24~36 h后达到峰值,比卵和若虫稍晚,可能是由于烟粉虱成虫体表有白色蜡粉等物质,延迟了球孢白僵菌对虫体的侵染。研究表明3个BtβGRPs蛋白可能参与了烟粉虱对真菌侵染的免疫反应,有可能成为烟粉虱生防的新靶标。     

高温胁迫下绿豆象内参基因筛选及其Hsp70基因的表达特性

为明确Hsp70蛋白在绿豆象Callosobruchus chinensis高温耐受性中的作用,该研究筛选并克隆β-actin、α-tubulin、β-tubulin、EF1-α、GAPDH、Hsc70、AK和RPL40八个候选内参基因,采用实时荧光定量PCR(quantitative real-time PCR,qPCR)技术测定其在绿豆象雌雄成虫体内的表达量,通过geNorm、BestKeeperr和NormFinde软件对这8个候选内参基因的表达稳定性进行评价,选择最适宜的内参基因对不同高温胁迫下绿豆象体内Hsp70基因的表达特性进行测定。结果表明,8个候选基因引物均有良好的特异性和引物扩增效率,引物扩增效率介于91.6%～108.1%之间。高温胁迫下,β-tubulin基因较其他7个候选基因有较小的平均变异度、稳定值和标准差,为最适宜的内参基因。39、42和45℃高温处理1 h后绿豆象雌雄成虫体内Hsp70表达水平较对照显著上调,当处理时间延长至3 h后,Hsp70基因的相对表达量较对照也有不同程度上调,但差异不显著,表明绿豆象Hsp70基因可以响应高温胁迫,Hsp70蛋白在绿...