Abstract:With the increasing serious problem of heavy metal pollution and the burgeoning cross subject of biological and chemical technique, the test methods of the heavy metal elements which were based on the biotechnology turned up. This article introduced the application and development of the methods such as Enzymatic analysis, Immunoassay, Biosensor in the heavy metal detection. And the merits and demerits and the development prospect of these technologies were discussed.

Abstract:A high efficient alginate-degrading strain was isolated from rotten seaweed using sodium alginate as the sole carbon source. Based on the physiological characteristics as well as 16S rDNA sequencing, the strain was identified as Cobetia sp. WG-007. The optimized fermentation medium was composed of sodium alginate 6.0 g/L, peptone 5.0 g/L, NaCl 20 g/L, K2HPO4 0.25 g/L. The strain was cultivated at 25 ℃ for 30 h under the optimized conditions and the alginate lyase activity was increased to 160.7 U/mL, which was 3.52-fold compared with the control.

Abstract:A codon-optimized gene(named Sygdh) encoding a glucose 1-dehydrogenase of Thermoplasma acidophilum was cloned into the plasmid pET28a and functionally expressed in Escherichia coli. The expressed enzyme was purified to homogeneity using Ni-NTA affinity chromatography with an apparent molecular weight of 41.0 kDa by SDS-PAGE analysis and characterized using the model substrate of D-glucose. The purified recombinant SyGDH was 4.5 U/mg at pH 7.5 and 45 ℃, and highly stable at pH 6.0~8.0 and 55 ℃ or below. It was activated by Zn2+, but strongly inhibited by Cu2+ and not significantly affected by other metal ions tested and EDTA. The Km and Vmax of recombinant SyGDH towards D-glucose were 28.2 mmol/L and 6.5 U/mg, respectively. The results showed that SyGDH exhibits much higher affinity with NADP+ than NAD+ and remarkably resists to several organic solvents. Furthermore, two recombinant strains E. coli BL21/pET28a-S1 and E. coli BL21/pET28a-Sygdh, which include recombinant NADPH-dependent carbonyl reductase and recombinant SyGDH, respectively, were mixed and used for the asymmetric reduction of Ethyl 4-chloroacetoacetate to Ethyl-4-chloro-3-hydroxybutyrate in the coenzyme cycle system. The product yield reached 99.0%, which is 3.11 fold as high as that of reaction without adding recombinant SyGDH strain, confirming that the recombinant SyGDH could be used as NADPH regenerator in biocatalysis oxidation and reduction system.

Abstract:This research aimed to investigate the effects of extract fraction 2(Fr.2) from Antrodia cinnamomea on the rat hepatic stellate cells(HSCs). A cultured rat HSC line CFSC-8B was stimulated by PDGF-BB in the presence or absence of Fr.2. Cell proliferation, cell migration, extracellular matrix(ECM) production and anti-fibrotic mechanism were studied. We found that CFSC-8B cell proliferation could be induced by 10 ng/mL PDGF-BB for 48 h. The IC50 value of Fr.2 was 56.46 μg/mL. Fr.2(25 μg/mL) significantly inhibited CFSC-8B cell proliferation and migration induced by PDGF-BB. Fr.2 also decreased α-SMA, Col 1, Col 3 and Fn mRNA expression. Mechanistically, Fr.2 negatively regulated p-ERK, p-JNK and p-P38 expression. Therefore, we provide evidence to demonstrate Fr.2 from A. cinnamomea inhibited HSCs proliferation, migration and ECM production induced by PDGF-BB, which in part, through negatively regulates MAPK signaling.

Abstract:Scanning Electron Microscope(SEM) and Laser Particle Size Analyzer(LPA), X-ray Diffraction(XRD) and Differential Scanning Calorimetry(DSC) were used to study the influence on the morphology, the diameter distribution, the crystal and the thermodynamic properties of the annealed corn starch granules. The results showed that annealing changed the surface of corn starch granules and made an increase in its micropores and hollows. Corn starch granules diameters were increased and the crystallites kept unchanged by means of annealing treatment. Annealing leads to elevation of corn starch gelatinisation temperatures and sharpening of the gelatinisation range. The solubility and swelling power of native corn starch and annealed corn starch increase when the temperature rises, however the solubility and swelling power of annealed corn starch(AC) are lower than those of native corn starch(NC) when they are at the same temperature conditions.

Abstract:To investigate the protective effects of resveratrol against oxidative damage induced by high glucose for different time in HepG2 cell. The cells were treated with 33 mmol/L glucose to induce oxidative damage and with different concentrations of Res 0.1, 1, 10 μmol/L for 24, 48, and 72 h. The cell viability was measured by MTT assay, DCFH-DA was used as fluorescence probe for observing ROS level in cells, superoxide dismutase(SOD), malondialdehyde(MDA), glutathione peroxidase(GSH-PX), total antioxidative capacity(T-AOC) were determined by using commercially available kits. Real-time PCR was used to determine the relative mRNA expression of nuclear factor-E2-related factor(Nrf2) and hemeoxygenase-1(HO-1). Results show that compared with high glucose damage group, after 48h, cell viability of Res groups was significantly increased(p<0.05), ROS level was significantly decreased(p<0.05). In Res groups, cell redox state was improved, 10 μmol/L Res group was significantly improved for 48 h(p<0.01), its Nrf2 HO-1 mRNA levels was significantly up-regulated(p<0.01). It is a significant protective effect of resveratrol against oxidative damage induced by high glucose in HepG2 and it works through scavenging free radicals, improving cell redox state and up-regulating Nrf2 HO-1 mRNA levels.

Abstract:To investigate the protect effects of thyroid hormone(T3) on HepG2 with oxidative damage. Establishing an oxidative stress model of HepG2 cells cultured in vitro, adding different concentrations of T3 for protection, determining the differentiation of cellular ROS, survival, antioxidant enzymes and related genes. Pretreatment of 10-9, 10-7, 10-5 mol/L T3 for 24 h, the level of ROS significantly reduced and the survival rate of cells was significantly increased(p<0.05)in cells of 50, 100, 200 μmol/L H2O2 processing respectively. While, 10-5 mol/L T3 increased the level of ROS. The amount of T3 can significantly reduce MDA content(p<0.05), increase T-AOC, GSH-Px and SOD activity of cells of H2O2 oxidative damage(p<0.05). But excess T3 increased MDA content and decreased the antioxidant activity. The results of gene expression showed that T3 pretreatment significantly increased PI3K, Nrf2 in the damaged cells(p<0.05). T3 can improve the redox state of HepG2 cells of H2O2 oxidative damage because of the effect on antioxidant enzyme activities and the expression of anti-oxidation genes and the effect is related to the dose and cell damage levels.

Abstract:Combine fluorescence PCR and specific antigen-antibody reaction, a rapid and sensitive detection method BPA in food contact materials was developed. After the optimization of the coating antigen concentration, the amounts of probe DNA, and the antibody concentration, BPA detection limit was low as 3.0 fg/g, and the linear concentration range was 0.01~10 pg/g. The recoveries of fruit drinks were 82.8%~115.2%. This method was specific, sensitive, and suitable for trace migration detection research of BPA and other small molecule chemicals.

Abstract:Based on sensory scores, effects of category and concentration of protease on the enzymolysis of Meretrix lusoria L. meat were studied. Orthogonal experiment was used to obtain the optimal two-step complex enzymolysis condition of Meretrix lusoria L. meat. The meat was hydrolyzed with compound protease(75 U/g) and neutral protease(75 U/g) at 45 ℃ and natural pH for 3 h; then flavor protease(50 U/g) at 50 ℃ and natural pH for 1 h. The Meretrix lusoria L. hydrolysate possessed light yellow color, fresh salty flavor, and clam characteristic aroma. The sensory score, EUC, amino nitrogen content, degree of hydrolysis(DH), enzymatic hydrolysis yield of the Meretrix lusoria L. hydrolysate and content of peptides were 5.5, 53.25 g MSG/L , 4.16 g/L, 35.3%, 36.4% and 21.8 g/L, respectively.

Abstract:In this manuscript, the porkballs were produced with meat, celery and starch as raw material, The process parameters of porkballs were optimized through orthogonal tests combining with fuzzy mathematics method. The results showed that the optimum proportions of celery meatballs were fat 20%, starch 20%, celery juice 60% and carrageenan 1.5%. The score of celery meatballs was 4.79 with this formula, and the sensory quality was excellent, which belonged to grade Ⅰ.

Abstract:Three kinds of methods for reconstruction of genome-scale metabolic network model, which were based on KEGG online database, Uniprot-MetaCyc databases and homologous alignment, have been studied for the process automation. Meanwhile, it has proposed an algorithm called Mahalanobis distance which was used to calculate the distance between reactions character frequency histogram. This algorithm can be used to the auto-integration of the draft model reactions and identify the core breakpoint. As an illustration example, the all automatic methods were implemented in the process of genome-scale metabolic reconstruction of Scheffersomyces stipitis, and confirming that these can improve the efficiency of model reconstruction.

Abstract:This study was undertaken to extract flavonoids from cherry leaves and to investigate the anti-oxidative activity in vitro. Total flavonoids were extracted from cherry leaves. The antioxidant activities of the flavonoids were evaluated by the scavenging power on free radicals ·OH and O2-· and inhibiting effects on erythrocytes hemolysis and lipid peroxidation of hepatic homogenate in vitro system. The flavonoids showed concentration-dependent scavenging power on ·OH and O2-·. The scavenging rates for ·OH and O2-·of 30 μg/mL flavonoids solution were 85.67% and 72.45% respectively, and the scavenging capacity for ·OH was stronger than Vitamin C. Cherry leaves flavonoids also showed inhibition on lipid peroxidation of hepatic homogenate and on hydrogen peroxide-induced erythrocyte hemolysis in a concentration-dependent manner. The inhibition rates were 79.82% and 75.54% in final concentration 30 μg/mL. Cherry leaves flavonoids have concentration-dependent antioxidant activity in vitro, which can be used as natural plant antioxidants or food additives for the development and utilization.

Abstract:To provide basic information for high gravity sugarcane fermentation, external ethanol was added to evaluate its impact on fructose and glucose fermentations by S.cerevisiae GJ2008. YPDF medium was used to simulate sugarcane juice to carry out ethanol fermentation at four different external ethanol concentrations ranging from 0% to 12% with an initial yeast number of 1.12×108 cells/mL, and the contents of fructose and glucose were determined by HPLC(High Performance Liquid Chromatography). Area under the curve (AUC) of fermentation of fructose and glucose was obtained using GraphPad Prism 5 software to analyze the discrepancy in fructose and glucose consumption. External ethanol addition had an important impact on fructose and glucose consumption, S.cerevisiae GJ2008 could utilize both sugars rapidly at the external ethanol concentration of less than 9% but with an obvious preference for glucose. Consumption of both sugars was severely inhibited when the external ethanol concentration increased up to about 12%, and the biomass could not be effectively produced. Fructose utilization was inhibited to a larger extent than glucose by external ethanol addition. The ethanol production was reduced by 19%~27% after the addition of external ethanol higher than 6%.

Abstract:By studying the natural fermentation of grape Xinjiang isolation and identification of 24 yeast, lipid production capacity of different strains to study. Getting them through high-yield lipase activity measured strain Y13. Through its research capacity fat enzymatic properties, the use of enzymatic properties further optimized fermentation conditions: optimum temperature is 39 ℃, the optimum pH is 7.0, Ca2+ can promote enzyme fermentation, the optimal incubation time is 65 h.

Abstract:A microwave digestion coupled plasma atomic emission spectroscopy(ICP-AES) method was presented for the determinations of Al, Zn, Cu, Cd, As, Cr and Pb in Porphyra haitanensis, and the samples were collected from a from in east china sea. The recovery rates of metals were between 97.8% and 104.9%, and actual applications demonstrate that this method is sensitive to achieve the determinations of metal content in Porphyra haitanensis. The results showed that an significant increasing trends were found in the metal contamination of Porphyra haitanensis during the past five years(P<0.05, RSD<1.89%), and different tenderness of the Porphyra haitanensis collected in 2012 showed an increasing contaiminations in the first＞the second＞the third＞the fourth cultivation duration.

Abstract:Maillard reaction products(MRPs) were prepared using glycine-fructose and arginine-fructose by model Maillard reaction. The change of ultraviolet-visible(UV-Vis) and fluorescence spectroscopy, the effects of amount of substance ratio of amino to carbonyl group, initial pH, temperature on browning of MRPs and pH change during reaction process were investigated. The scavenging action of MRPs on DPPH and hydroxyl free radical, H2O2 and total reducing power were also observed. The results showed that UV absorption peaks were 220 and 266 nm, 220 nm for glycine-fructose and arginine-fructose, respectively, excitation and emission wavelengths for fluorescence peaks were 339 and 441 nm, 339 and 426 nm, respectively. The browning and acidity of MRPs all increased with the heating time. Browning all reaches their corresponding maximum when the amount of substance ratio of amino to carbonyl group was 1∶3, pH 11～12 and temperature 110～120 ℃, respectively. With the increasing of MRPs concentration, scavenging action on hydroxyl free radical increased to a maximum value and then decreased, H2O2 and total reducing power showed a dose-response relationship to some degree, and scavenging action on DPPH was lower and decreases with increasing concentration.

Abstract:Nano-selenium was prepared with locust bean polysaccharides as soft template by the reduction of ascorbic acid(Vc) to selenious acid and characterized by UV and SEM. The effects of reaction time, temperature, locust bean polysaccharide and Vc concentration on particle size and morphology of the resulting product were investigated. The results showed that uniformly red globular nano-selenium was obtained with average diameter about 95～100 nm when the mass fraction of locust bean polysaccharide was 0.2‰ and the molar ratio of ascorbic acid to selenite 5∶1 at 60 ℃ for 4 h.

Abstract:This research was aimed to compare the role of anti-Saccharomyces cerevisiae of garlic extract by using ultrasonic assisted extraction method for different extraction under different conditions. The optimum extraction and determination of the minimum inhibitory concentration and minimum bactericidal concentration were preferably tested. The results showed that: the antibacterial activity of garlic extract under optimum conditions when Garlic extract with 30% ethanol as solvent, as well as Ultrasonic assisted extraction at 40 ℃ extraction 3 h, extraction times of 3, and solid-liquid ratio of 1∶5. Under this condition, garlic beer TomatoLateBlightStrain bacteriostatic diameter of 37.2 mm. Garlic extract on the minimum inhibitory concentration of S. cerevisiae 12.5 mg/mL, minimum bactericidal concentration of 25 mg/mL.