Cloning,sequencing and structural analysis of membrane-bound polyphenol oxidase from Granny Smith apples (Malus × domestica Borkh)

The gene sequence coding for the membrane-bound polyphenol oxidase (mPPO) with a length of 1761 bp was cloned by PCR method and shown to contain one highly conserved sequence encoding a di-copper-binding region. The predicted three-dimensional structure of mPPO indicated that the active site was located near two copper ions and composed of a typical bundle of four α-helices. Each of the two catalytic copper ions was coordinated with three histidine residues in the hydrophobic pocket, yielding His 180, His 201, His 210, His 332, His 336 and His 366. Docking studies showed that 4-methylcatechol and chlorogenic acid have different binding models due to different ligand sizes and binding sites in the active centre, and it was found that the smaller compound exhibited a higher affinity for mPPO. Molecular dynamic simulation results indicated that Phe 353 is important in controlling enzymatic activity through influencing substrate coordination in the active site.     

Optical studies of Y_3(Al,Ga)_5O_(12):Ce~(3+),Cr~(3+),Nd~(3+) nano-phosphors obtained by the Pechini method

The Y3(AI,Ga)_5O_(12):Ce~(3+),Cr~(3+),Nd~(3+)(YAGG) nano-phosphors with homogeneous particle-size distribution,low aggregation and average crystalline size of about 65 nm were obtained using a modified Pechini method.Only slight aggregation of the crystallites occurs after post-annealing at 1100℃.The intense Ce~(3+)bands in the excitation spectra of the Ce~(3+),Cr~(3+),Nd~(3+)co-doped materials monitoring the Cr~(3+) emission at 690 nm indicate energy transfer from Ce~(3+) to Cr~(3+).Weak Nd~(3+) lines are observed,as well.In addition,the emission of Nd~(3+)at 1060 nm with excitation of Ce~(3+) and Cr~(3+) confirms the Ce~(3+)/Cr~(3+)to Nd~(3+)energy transfer.The short average luminescence decay times for the Ce~(3+) emission indicate the Ce~(3+)/Cr~(3+)to Nd~(3+)energy transfer.Eventually,the Y_3(AI,Ga)_5O_(12):Ce~(3+),Cr~(3+),Nd~(3+) nano-phosphors exhibit persistent luminescence originating from the 4f~3→4f~3 transitions of Nd~(3+) which matches well to the first biological window to be used in bioimaging applications.     

CaDHN3, a Pepper (Capsicum annuum L.) Dehydrin Gene Enhances the Tolerance against Salt and Drought Stresses by Reducing ROS Accumulation

Dehydrins (DHNs) play an important role in abiotic stress tolerance in a large number of plants, but very little is known about the function of DHNs in pepper plants. Here, we isolated a Y₁SK₂-type DHN gene “CaDHN3” from pepper. To authenticate the function of CaDHN3 in salt and drought stresses, it was overexpressed in Arabidopsis and silenced in pepper through virus-induced gene silencing (VIGS). Sub-cellular localization showed that CaDHN3 was located in the nucleus and cell membrane. It was found that CaDHN3-overexpressed (OE) in Arabidopsis plants showed salt and drought tolerance phenotypic characteristics, i.e., increased the initial rooting length and germination rate, enhanced chlorophyll content, lowered the relative electrolyte leakage (REL) and malondialdehyde (MDA) content than the wild-type (WT) plants. Moreover, a substantial increase in the activities of antioxidant enzymes; including the superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), ascorbate peroxidase (APX), and lower hydrogen peroxide (H₂O₂) contents and higher O₂^•− contents in the transgenic Arabidopsis plants. Silencing of CaDHN3 in pepper decreased the salt- and drought-stress tolerance, through a higher REL and MDA content, and there was more accumulation of reactive oxygen species (ROS) in the CaDHN3-silenced pepper plants than the control plants. Based on the yeast two-hybrid (Y2H) screening and Bimolecular Fluorescence Complementation (BiFC) results, we found that CaDHN3 interacts with CaHIRD11 protein in the plasma membrane. Correspondingly, the expressions of four osmotic-related genes were significantly up-regulated in the CaDHN3-overexpressed lines. In brief, our results manifested that CaDHN3 may play an important role in regulating the relative osmotic stress responses in plants through the ROS signaling pathway. The results of this study will provide a basis for further analyses of the function of DHN genes in pepper.     

Protein quality and antioxidant properties of soymilk derived from black soybean after in vitro simulated gastrointestinal digestion

Protein quality and antioxidant properties of soymilk derived from black soybean (eight varieties) in China were analysed following in vitro simulated gastrointestinal digestion (including dialysis). Soymilk from black soybean possessed high okara weight but low yield, protein content and sensory scores. The in vitro digestibility of protein in all black soymilk samples was higher than 60%, and the Shenmu black soybean exhibited the highest digestibility. Non-digested milk from the black soybean exhibited significantly high total phenolic content (TPC) (127.15–173.04 mg/100 mL), ferric-reducing antioxidant power (FRAP) (272.18–366.27 μmol L⁻¹) and DPPH free radical-scavenging activity (61.20–83.81%). These parameters were significantly lower in the non-digested soymilk than those in soymilk after gastric digestion but higher than those of soymilk in the dialysed fraction. Gastric digestion significantly increased bioactive compound levels released from black soymilk, and the bioaccessibility of phenolic compounds was 24.37–36.05%. Hence, black soymilk was sufficiently available for human absorption.     

Bio-economic modelling for marine spatial planning application in North Sea shrimp and flatfish fisheries

Marine activities have been increasingly competing for space and reducing areas for fishing. The use of spatially explicit tools can assist the decision making process on defining the optimal location of closures for fishing due to these emerging activities.This paper presents the Spatial Integrated bio-economic Model for FISHeries (SIMFISH). In this model fishers behaviour is simulated based on optimal effort allocation. The added value of this model compared to other existing spatial management tools lies in the presence of (i) short and long term fishers behaviour (ii) spatial explicit stock and fleet dynamics and (iii) relatively low data requirements.As an illustration, SIMFISH is applied in this paper to estimate the impact of area closures in the North Sea. Overall area closures have a negative impact on the profitability of the fleets. This would be worsened with higher fuel prices and decreased stock productivity.     

Effects of size and autoclavation of fruit and vegetable wastes on biohydrogen production by dark dry anaerobic fermentation under mesophilic condition

Fermentative hydrogen production from fruit and vegetable wastes (FVWs) through Dry Fermentation Technology (DFT) was studied through three independent experiments in order to find out the effect of particle size and autoclaving pretreatment on bio-hydrogen production from FVWs and as follows: (1) autoclaved FVWs with sizes < 5 cm (experiment I); (2) raw FVWs with sizes < 5 cm (experiment II) and (3) autoclaved FVWs with sizes > 5 cm (experiment III). The assay with autoclaved waste yielded a higher percentage of hydrogen in the headspace of the dry fermenter reaching a maximum value of 44% in experiment I. However, the maximum hydrogen production was obtained in experiment III with 14573 NmL at a yield of 23.53 NmL H₂/gVS. Profiling of the microbial communities by denaturing gradient gel electrophoresis (DGGE) indicated that the most prominent species were the genera Clostridium, Bifidobacterium, and Lactobacillus.     

Comparison of biochemical properties and thermal inactivation of membrane‐bound polyphenol oxidase from three apple cultivars (Malus domestica Borkh)

A comparative study of the properties of membrane‐bound polyphenol oxidase (mPPO) from three apple cultivars, namely Red Fuji (FJ), Granny Smith (GS) and Golden Delicious (GD), was carried out for the first time. Data indicate that mPPOs from three cultivars exhibit significantly different properties. GS mPPO had the strongest affinity to catechol, but FJ mPPO had the highest maximum velocity. Red Fuji (FJ) mPPO had the significantly higher activity than those of GD and GS mPPOs. Red Fuji (FJ) mPPO had the highest activity at pH 8.00, while GD and GS mPPOs at 4.50 and 7.50–8.00, respectively. Red Fuji (FJ) mPPO was more stable than GD and GS mPPOs over the pH range of 5.0–8.5. The optimal temperature for GS mPPO was within 70–75 °C, which is higher than those for mPPOs from FJ and GD. Thermal inactivation of the three mPPOs followed a first‐order kinetic model with different inactivation kinetic parameters.     

基于CT图像的细观混凝土孔隙缺陷研究

将混凝土看作由骨料、砂浆、界面和孔隙缺陷组成的细观非均质材料,借助计算机断层扫描(CT)技术,建立了考虑孔隙缺陷相的细观混凝土模型,对有、无孔隙缺陷情况下混凝土的受拉破坏过程及应力集中的大小与分布进行了对比分析,研究了孔隙缺陷的面积统计分布规律.结果表明:孔隙缺陷的存在会增强混凝土材料的整体非均质性和拉应力分布的非均匀性,导致明显的应力集中;孔隙缺陷面积的分布规律基本符合Lognormal概率分布规律.