Electrochemical characteristics of heme proteins in hydroxyethylcellulose film

A stable film made from hydroxyethylcellulose (HEC) on pyrolytic graphite (PG) electrode was employed for incorporating hemoglobin (Hb), myoglobin (Mb) and horseradish peroxidase (HRP), and the electrochemical characteristics of the proteins were studied correspondingly. Experimental results revealed that HEC film could greatly accelerate electron transfer between the proteins and electrode, and the proteins showed a thin layer electrochemical behavior in the film. Moreover, all the proteins in the film exhibited good catalytic activity towards the reduction of hydrogen peroxide (H₂O₂) in the low H₂O₂ concentration range. In the high concentration range, H₂O₂ would exhibit toxicity effect on the proteins. The electrochemical properties and electrocatalytic abilities of the three heme proteins in HEC film have been compared, and the optimal conditions for H₂O₂ biosensor fabrication have been obtained.     

Characterization of a Thermostable and Surfactant-Tolerant Chondroitinase B from a Marine Bacterium Microbulbifer sp. ALW1

Chondroitinase plays an important role in structural and functional studies of chondroitin sulfate (CS). In this study, a new member of chondroitinase B of PL6 family, namely ChSase B6, was cloned from marine bacterium Microbulbifer sp. ALW1 and subjected to enzymatic and structural characterization. The recombinant ChSase B6 showed optimum activity at 40 °C and pH 8.0, with enzyme kinetic parameters of K_m and V_max against chondroitin sulfate B (CSB) to be 7.85 µg/mL and 1.21 U/mg, respectively. ChSase B6 demonstrated thermostability under 60 °C for 2 h with about 50% residual activity and good pH stability under 4.0–10.0 for 1 h with above 60% residual activity. In addition, ChSase B6 displayed excellent stability against the surfactants including Tween-20, Tween-80, Trion X-100, and CTAB. The degradation products of ChSase B6-treated CSB exhibited improved antioxidant ability as a hydroxyl radical scavenger. Structural analysis and site-directed mutagenesis suggested that the conserved residues Lys248 and Arg269 were important for the activity of ChSase B6. Characterization, structure, and molecular dynamics simulation of ChSase B6 provided a guide for further tailoring for its industrial application for chondroitin sulfate bioresource development.     

Biochemical characterisation of an aminopeptidase with highest preference for lysine from Japanese flounder skeletal muscle

An aminopeptidase was purified from Japanese flounder skeletal muscle to homogeneity by ammonium sulphate fractionation and three chromatographies. The enzyme was approximately 100 kDa with isoelectric point of 5.7 as estimated by two-dimensional polyacrylamide gel electrophoresis. Its optimum temperature and pH were 45 °C and 7.5, respectively. According to peptide mass fingerprinting study, the enzyme revealed high identity to a puromycin-sensitive aminopeptidase. It had a broad specificity toward aminopeptidase substrates and preferred to hydrolyse Lys-MCA with k_cat/K_m of 8.1 × 10⁶ M^?1 s^?1, and the activation energy (E_a) of 72.5 kJ M^?1. Metal-chelating agents effectively inhibited the enzyme activity, and Zn²⁺, Mn²⁺ and Co²⁺ significantly restored the apoenzymatic activity dialysed by EDTA, whilst inhibitors to other proteinases did not show much effect. Furthermore, bestatin strongly inhibited its activity. These results indicate that the purified enzyme is a metalloaminopeptidase which would possibly contribute to free amino acids increase in fish muscle.     

Ca2+ enhances algal photolysis hydrogen production by improving the direct and indirect pathways

In this study, Ca²⁺ was applied to enhance the photolysis H₂ production of Chlamydomonas reinhardtii grown in pure culture or co-cultivated with bacterium. With addition of Ca²⁺, the chlorophyll and protein contents in algal cells were maintained at high levels and the reactive oxygen species (ROS) were scavenged to tolerable levels by activities of catalase (CAT) and superoxide dismutase (SOD). High chlorophyll and low ROS are benefit to protect the photosystem II (PS II) activity that responsible for the direct photolysis H₂ production. Moreover, addition of Ca²⁺ was able to promote the acetic acid assimilation as well as the starch synthesis. The starch degradation, in return, contributed to the H₂ production via the indirect photolysis pathway. Based on the improvements of direct and indirect photolysis pathways, the H₂ production of algae grown in co-cultivation adding with 5 mM Ca²⁺ reached a maximum value of 306 mL L⁻¹, which was 2.4 times higher than the production without Ca²⁺ addition. This study gives an efficient and easy strategy to improve the algal photolysis H₂ production, and gets an insight into the mechanisms of the improvement.     

The effect of glaze uptake on storage quality of frozen shrimp

Ice-glazing is applied to protect the frozen shrimp from undesirable quality changes during frozen storage. Effects of initial frozen shrimp temperature on glaze uptake; glazing time on glaze uptake; and different glaze percentage on physical and chemical changes of frozen shrimp during storage were investigated. Shrimps were frozen in a spiral freezing machine (?35 °C/15 min); transferred to the air blast freezer until the core temperature reached ?18 °C, ?25 °C and ?30 °C; submitted to glazing process; and stored at ?18 °C for 180 days. The glazing percentage, pH and N-TVB levels were monitored every 45 days. This study has demonstrated the effectiveness of the glazing process as a protecting agent for frozen shrimp. A reasonable range of water uptake could be between 15% and 20% to guarantee the final quality. Therefore, it is important to prevent temperature fluctuations during transportation and storage to maintain the quality of the frozen shrimps.     

Purification and characterization of parvalbumins from silver carp (Hypophthalmichthy molitrix)

BACKGROUND: As the largest producer and consumer of freshwater fish in the world, many people suffer from allergy for consuming freshwater fish in China. However, the allergen profiles of freshwater fish are rarely known. RESULTS: Parvalbumins (PVs) from the white muscle of silver carp (Hypophthalmichthy molitrix) were purified by ammonium sulfate fractionation and column chromatography including DEAE‐Sepharose and Superdex 75. Three PV isoforms—PV‐I, PV‐II, and PV‐III—were obtained and their molecular masses as estimated by tricine–sodium dodecyl sulfate–polyacrylamide gel electrophoresis were 12, 11, and 14 kDa, respectively. All the PVs could be detected by anti‐frog PV monoclonal antibody. PV‐I and PV‐II were quite possibly glycoproteins, while PV‐III was not glycosylated, as analyzed by periodic acid–Schiff (PAS) staining. Thermal stability revealed that PV‐I and PV‐II easily formed polymers, while these proteins were stable in a pH range of 4.0–10.0. A PV gene encoding 110 amino acid residues was cloned and it revealed high identity with PVs from other species of fish. CONCLUSION: Three isotypes of PV were purified to homogeneity and one distinct PV gene was cloned in silver carp white muscle. Copyright © 2010 Society of Chemical Industry     

Radio frequency combined hot air (RF-HA) drying of tilapia (Oreochromis niloticus L.) fillets: Drying kinetics and quality analysis

Hot-air assisted radio frequency (RF-HA) heating was utilized for tilapia fillets drying to achieve rapid, uniform drying and improved sample physical properties. Effects of sample thickness (5, 8 and 10 mm) and with or without wrapping material (skin or gauze) were investigated under both HA and RF-HA drying treatments. The drying characteristics and quality parameters of tilapia fillets in hot air (HA) and RF-HA drying were then determined. Results show the drying rate of RF-HA drying is 1.1–1.4 times of that of HA drying for tilapia samples with different thicknesses. RF-HA dried samples are with higher rehydration rate and lower volume shrinkage rate, especially for fillets with larger thickness. The wrapped gauze did not show significant influence on the color of tilapia sample in RF-HA drying but improved the rehydration rate, while the samples with preserved fish skin resulted in a lower overall quality under both HA and RF-HA drying treatments. Page model and Henderson-Pabis models were suitable to be used describing the drying kinetics of RF-HA drying and HA drying tilapia, respectively.     

基于乙酰胆碱酯酶和氧化应激研究海胆酮对阿尔茨海默症的作用机制

海胆酮是一种酮式类胡萝卜素,主要从海胆及藻类等海洋生物中提取。本文研究海胆酮对乙酰胆碱酯酶（acetylcholinesterase,AChE）的抑制作用,应用酶动力学、荧光光谱、圆二色光谱和分子对接技术研究海胆酮对AChE的抑制机理,并用淀粉样β蛋白片段25～35（amyloid beta-peptide 25-35,Aβ25-35）诱导大鼠肾上腺嗜铬细胞瘤细胞（PC12细胞）建立阿尔茨海默症（Alzheimer’s disease,AD）模型,研究海胆酮对AD细胞模型氧化应激损伤的作用。结果表明,海胆酮有很强的AChE抑制活性,其半抑制质量浓度为（16.29±0.97）μg/mL,抑制常数Ki为3.82 μg/mL,表现为竞争性抑制;海胆酮可诱导AChE二级结构改变,更容易与AChE活性中心氨基酸Ser200、His440、Trp84和Tyr121结合,阻碍底物碘代硫代乙酰胆碱（acetylthiocholine iodide,ATCI）与酶结合,从而引起酶活力降低。海胆酮能有效抑制Aβ25-35诱导PC12细胞的AChE活力,降低丙二醛含量,增加超氧化物歧化酶、过氧化氢酶和谷胱甘肽过氧化物酶活力,减轻Aβ25-35诱导的PC12细胞氧化应激损伤。本研究基于AChE和氧化应激阐明了海胆酮对AD的潜在作用机制,为海胆酮在功能食品、生物医药等领域的应用提供了数据支持和理论根据。     

魔芋胶对南美白对虾肌原纤维蛋白凝胶特性的影响

为探究魔芋胶（konjac glucomannan,KGM）对南美白对虾肌原纤维蛋白（shrimp myofibrillar protein,SMP）凝胶性能的影响,将KGM与SMP按不同比例（1∶50、1∶20、1∶10）进行复配制备复合凝胶体系SK50、SK20、SK10,通过测定表面疏水性、内源性荧光、浊度及粒径、流变学、红外光谱、蛋白变化及凝胶微观结构,研究复合体系的凝胶特性变化。结果表明,SMP及SMP-KGM复合体系的表面疏水性、浊度及粒径随着KGM添加量的增大而增大,流变学分析表明SMP及SMP-KGM复合体系均出现剪切稀化现象,且SK20的G’值最高,其凝胶的结构稳定性最好。红外光谱分析表明SMP及SMP-KGM复合体系组的光谱特征带相似,表明无明显的基团生成。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析表明KGM的加入并没有引起蛋白条带的明显变化,相较于未加热组,加热后样品中肌球蛋白重链、副肌球蛋白以及肌动蛋白的条带明显减弱,说明加热能促使三者发生热聚合反应。扫描电镜与分形维数分析表明,随着KGM添加量的增加,形成了致密有序的凝胶。结果表明,一定量的KGM添加能有效提升虾糜的凝胶特性,提高虾糜类产品品质。     

贮存条件对Gairdner大麦和麦芽的醛类物质的影响

利用固相微萃取与气相色谱-质谱联用检测醛类物质的方法跟踪了啤酒酿造大麦和麦芽Gairdner中的脂质氧化醛、Strecker醛、糠醛在10℃和35℃条件下的变化趋势。新鲜麦芽中的Strecker醛和糠醛含量高于大麦,而脂肪氧化醛含量和脂合氧化酶活力明显低于大麦。大麦和麦芽中脂质氧化醛和糠醛在贮存过程明显增加,具有指示大麦和麦芽新鲜度的可能性。10℃和35℃贮存5个月后,大麦的反-2-壬烯醛分别增加了1.01和1.62倍,而麦芽的反-2-壬烯醛分别增加5.12和17.74倍。由于反-2-壬烯醛的高风味活性特点,建议将反-2-壬烯醛作为大麦和麦芽新鲜度与品质优劣的关键性评价指标。本研究从脂质氧化醛等指标量化的角度,再次肯定了低温贮存相对于高温贮存更利于大麦和麦芽保持新鲜的事实。