用电子束辐射法制备固定化乳酸脱氢酶

前言固定化酶的制备,近年来已有许多进展.固定化技术之一的凝胶格子包埋法,尤其受到重视,因为它与其它方法不同,酶分子本身在固定化过程中几乎完全不发生结合或偶联反应,而是被包埋在凝胶的微细格子里.这样,它可能对酶蛋白的构象。

大米饮料制备工艺研究

利用糙米和精米经烘炒、生物酶解、过滤、罐装、包装等工序制造色泽清亮、香气宜人、保留糙米和精米原有成分的甘甜可口的大米饮料。对烘烤时间及水解条件等关键工序作了较为系统的研究,大米最佳烘烤条件:白米190℃、20min效果较好,糙米180℃、20min。最佳水解条件为蛋白水解酶2%,葡萄糖化酶125U/g,温度50℃,时间7h。

Genome-wide Analysis of Glucose-6-phosphate Dehydrogenase(G6PDH) and Its Evolution in Eucalyptus grandsis

[Objective] The aim of this study was to perform genome-wide analysis of glucose-6-phosphate dehydrogenase（G6PDH） and reveal its evolution in Eucalyptus grandsis.[Method] The gene character,protein sequence and phylogenetic tree of G6PDH gene were analyzed by BLAST and other bioinformatics software within Eucalyptus grandsis whole genome database.[Result] Six G6PDH genes,including one cytomic type and five plastids,were detected in the E.grandsis genome.All the G6PDHs have conserved motifs of motif 1,motif 2,motif 3,motif 7,motif 9 and motif 11.Furthermore,promoter sequences of all E.grandsis G6PDH contain TATA box,enhancer,light-responsive,hormone-responsive and stress-responsive regulatory elements.[Conclusion] This study provided reference for the further revealing molecular function of E.grandsis G6PDH gene family

Genetic improvement of Trichoderma ability to induce systemic resistance

The beneficial applications of Trichoderma spp. in agriculture include not only the control of plant pathogens, but also the improvement of plant growth, micronutrient availability, and plant tolerance to abiotic stress. In addition, it has been suggested that these fungi are able to increase plant disease resistance by activating induced systemic resistance (ISR) . The mode of action of these beneficial fungi in the Trichoderma -plant-pathogen interaction are many, complex and not completely understood. Numerous lytic enzymes have been characterized, the encoding genes (ech42 gluc78, nag1 from T. atroviride strain P1) cloned, and their role in biocontrol demonstrated. The corresponding biocontrol-related inducible promoters have been used in a reporter system based on the Aspergillus niger glucose oxidase gene (goxA) to monitor biocontrol activity. Glucose oxidase catalyzes the oxygen-dependent oxidation of D-glucose to D-glucono-1,5-lactone and hydrogen peroxide; this latter compound is known to have an antifungal effect and activate the plant defence cascade, thus increasing resistance to pathogen attack. T. atroviride P1 transformants with various promoters gox were tested as seed coating treatments on bean seeds planted in soil infested with a soilborne fungal pathogen. Successively, the emergent leaves were inoculated with a foliar pathogen to determine the effect of the GOX transformants on biocontrol and resistance to pathogen attack. Inoculations with the P1-GOX transformants not only reduced disease symptoms caused by a soil pathogen, but also the lesions of various foliar pathogens applied far from the Trichoderma colonization, thus activating ISR. A similar approach is being use to genetically improve T. harzianum T22, a rhizosphere competent and commercially marketed strain not transformed yet, by using four different gox gene constructs under the control of constitutive and inducible promoters. Plasmids have been introduced in Trichoderma by protoplasts co-transformation. hygromicin resistant progeny selected, and mitotically stable transformants analysed to confirm the presence of the novel enzyme activity. Progenies are being tested for biocontrol ISR inducing activity.

Polyaniline-graphite composite film glucose oxidase electrode

A novel polyaniline-graphite composite film glucose oxidase （PGCF GOD） electrode was developed. The PGCF was synthesized by cyclic voitammetry method in 0.5 mol/L H2SO4 solution containing 1 g/L graphite powder and 0.2 mol/L aniline. The PGCF GOD electrode was prepared by doping GOD into the composite film. The morphology of the PGCF and the response property of the PGCF GOD electrode were investigated by scanning electron microscopy and electrochemical measurement, respectively. The results show that the PGCF has a porous and netty structure and the PGCF GOD electrode has excellent response property such as high sensitivity and short response time. Influences of pH value, temperature, glucose concentration and potential on the response current of the electrode were also discussed. The sensor has a maximum steady-state current density of 357.17μA/cm2 and an apparent Michaelis-Menten constant of 16.57 mmol/L. The maximum current response of the enzyme electrode occurs under the condition ofpH 5.5, 0.8 V and 65℃.

Bienzyme system immobilized in biomimetic silica for application in antifouling coatings

Antifouling coatings are used extensively on vessels and underwater structures. Conventional antifouling coatings contain toxic biocides and heavy metals, which may induce unwanted adverse effects such as toxicity to non-target organisms, imposex in gastropods and increased multiresistance among bacteria. Therefore,enzyme-based coatings could be a new alternative solution. A H2O2-producing bienzyme system was developed in this study. H2O2 can be produced from starch by the cooperation of α-amylase and glucose oxidase, which promotes the hydrolysis of polymeric chain and oxidizes the glucose to produce H2O2, respectively. The encapsulated bienzyme(A-G@BS) exhibits enhanced stabilities of thermal, pH, recycling and tolerance of xylene. The A-G@BScontaining coating releases H2O2 at rates exceeding a target of 36 nmol·cm-2·d-1for 90 days in a laboratory assay. The results demonstrate that the method is a promising coating technology for entrapping active enzymes,presenting an interesting avenue for enzyme-based antifouling solutions.

1,2,3,4,6-penta-O-galloyl-β-D-glucose up-regulates heme oxygenase-1 expression by stimulating Nrf2 nuclear translocation in an extracellular signal-regulated kinase-dependent manner in HepG2 cells

AIM: To examine the potency of 1,2,3,4,6-penta-O-galloyl-β-D-glucose (PGG) as a hepatic heme oxygen-ase-1(HO-1) inducer and its regulation in HepG2 cells. METHODS: Expression of HO-1 and NF-E2-related factor 2 (Nrf2) and activation of mitogen-activated protein (MAP) kinases were analyzed by Western blot, immuno-fluorescence assay, and flow cytometry. Transfections of HO-1 gene, small interfering RNAs for HO-1 and Nrf2, and dominant-negative gene for MAP/extracellular signal-regulated kinase (ERK) were carried out to dissect the signaling pathways leading to HO-1 expression in HepG 2 cells. RESULTS: PGG up-regulated HO-1 expression and this expression conferred cytoprotection against oxidative injury induced by t-butyl hydroperoxide. Moreover, PGG induced Nrf2 nuclear translocation, which was found to be an upstream step of PGG-induced HO-1 expression, and ERK activation, of which pathway was involved in PGG-induced Nrf2 nuclear translocation, HO-1 expression and cytoprotection. CONCLUSION: PGG up-regulates HO-1 expression by stimulating Nrf2 nuclear translocation in an ERK-depen-dent manner, and HO-1 expression by PGG may serve as one of the important mechanisms for its hepatoprotective effects.

Macroalage as a source of alpha-glucosidase inhibitors

Alpha-glucosidase inhibitors were screened from organic solvent extracts of macroalgae by a spec- trophotometrical method with p-nitrophenyl-D-glucopyranosidase as the substrate. The result indicates that or- ganic crude extracts from some macroalgae such as Rhodomela confervoides (Huds.) Silva, Gracilaria textorii (Suringar) DeToni, Plocamium telfairiae Harv., Dictyopteris divaricata (Okam.) Okam, Ulval pertusa and En- teromorpha intestinalis (L.) Link et al. show strong inhibitory activity of alpha-glucosidase at concentration of 79.6 μg/ml.

Verification of protein sparing by feeding carbohydrate to common carp Cyprinus carpio

A 9-week feeding trial in floating freshwater cages(1.0 m×1.0 m×2.0 m) was conducted to study the effects of different dietary levels of protein and starch on growth,body composition,and gene expression of enzymes in common carp,Cyprinus carpio(mean body weight,36.12±1.18 g) to evaluate the protein-sparing effect of dietary carbohydrate.Four diets were formulated with corn starch as the carbohydrate source to obtain corn starch levels of 6.5%,13%,19.5%,or 26%and protein levels of 30.5%,28.2%,26.4%,and 24.2%.The results showed no differences in growth performance of fish fed the diets with different protein and corn starch levels,but body composition and glucose metabolic enzyme activity of carp were significantly affected by the different diets(P<0.05).Weight gain,specific growth rate,and the feed conversion ratio were not different in fish fed the different dietary treatments.Protein efficiency ratio increased significantly as corn starch level increased(P<0.05).Whole-body crude lipid composition increased with increasing dietary corn starch level(P<0.05).Glucokinase(GK),hexokinase,and pyruvate kinase(PK) activities increased significantly with increasing dietary corn starch level(P<0.05),whereas glucose-6-phosphate(G6Pase) activity decreased with increasing dietary corn starch level(P<0.05).GK gene expression was significantly higher in fish fed the high-corn starch diet than those fed the low-corn starch diet(P<0.05).G6 pase gene expression tended to decrease with increasing starch level(P>0.05).In summary,the results indicate a protein-sparing effect by substituting carbohydrate in the diet of common carp.

Membrane Microfiltration Fermentation of Glucose Oxidase with Cell Recycling

Membrane microfiltration fermentation (MMF) with cell recycling was successfully applied to the production of glucose oxidase (GOD). A plate microfiltration module was found suitable for such purpose. By feeding whole medium in MMF, the productivity of GOD was much higher than that by feeding glucose alone. With increasing dilution rate the enzyme productivity increased and average enzyme activity decreased. The enzyme productivity of MMF under D = 0.12h-1 and 0.20h-1 were 3871 and 3945U·h-1 respectively, which was about 3 times as that of batch fermentation (BF) and the average enzyme activity was still as high as STU·mL-1 under D = 0.12h-1. The relative efficiency of MMF applied to low yield strain was higher than that applied to high yield strain.

Effect of puerarin on the P13K pathway for glucose transportation and insulin signal transduction in adipocytes

To explore the effect of puerarin on insulin receptor （IR）, insulin receptor substrate-1 （IRS-1） and protein expression of protein kinase B （PKB） in the P13K pathway of the glucose consumption, transportation and insulin signal transduction in 3T3-L1 adipoeytes with insulin resistance. The insulin resistance 3T3-L1 adipocytes model was established by free fatty acid induction. The model cells were managed with puerarin in different concentrations. Glucose consumption was detected with glucose oxidase method, glucose transportation rate was determined by 2-deoxy-^3H glucose ingesting method, and the IR, IRS-1 and PKB expression were determined by Western blot. Glucose consumption and transportation were significantly decreased in the model adipoeytes, but increased after treated with puerarin （P 〈 0. 01 ）. Moreover, the level of tyrosine phosphorylation of IR subunit β was higher in the puerarin treated groups, and that of IRS-1 was higher in the group treated with low dose puerarin than that in the model group. The 3T3-L1 adipocytes of insulin resistance model could be induced by free fatty acid successfully, puerarin could promote the glucose utilization in them to alleviate the in- sulin resistance, which may be related with the action in advancing the tyrosine phosphorylation of IR and IRS-1.

Coronatine Induces an Accumulation of Anthocyanin and Starch in Purple-fleshed Sweetpotato(Ipomoea batatas Lam.)

[Objective] The objective of this research was to examine the effects of COR on anthocyanin and starch content in storage roots of two PFS genotypes, and to explore the relationships between anthocyanin synthesis and starch accumula- tion. [Method] A field experiment was carried out to determine the changes in yielc components, yield, contents of anthocyanin and starch, activities of phenylalanine ammonia-lyase （PAL） and adenosine 5-diphosphate glucose pyrophosphorylase （AG- Pase） in two genotypes of PFS （Ipomoea batatas L., var. ＇Ayamurasaki＇ and ＇Jishu18＇）. [Result] The application of COR significantly increased starch and antho- cyanin content in storage roots of Jishu18 across developmental stages by inducing the activities of PAL and AGPase, and finally enhanced yield by promoting fresh weight of storage roots. Ayamurasaki was insensitive to treatment with COR al- though its PAL activity temporally increased. The starch and anthocyanin content of Aya, and the anthocyanin content of Jishu18 increased progressively across devel- opmental stages with or without COR application, but the starch content of Jishu18 increased initially, then decreased before increasing again without application of COR. Treatment with COR reduced downward trend of starch accumulation in Jishu18. Thus, the effect of COR on accumulation of anthocyanin and starch in storage roots of PFS differs according to genotypes. [Conclusion] The application of 0.05 μmol/L COR may increase starch and anthocyanin content in PFS genotypes with lower starch and anthocyanin content in storage roots.

Kinetic Parameter of Alfa-amylase and Glucoamylase Enzymatic Reaction on the Glucose Yield from Hydrolyzed Processes of Tapioca Solid Waste

The increasing of tapioca production nowadays effected the production of waste. The waste of tapioca industries consists of two kinds, which were liquid waste and solid waste. Further more, tapioca solid waste treatment was ineffective. Weather solid waste produced from the extraction process still contains high concentration of starch that can be used to produce high quality product, for example, bio ethanol or other alternative energy sources. Objective of these experimental work was utilizing solid waste of tapioca industries and looking for the exactly composition of n-amylase and gluco-amylase enzymes on the hydrolysis processes of the solid waste of tapioca. The exact composition from both enzymes can be expected to increase the yield of glucose. Variables of cx-amylase enzyme for this research were 0.3% （w/w） and 0.5% （w/w） with liquefaction time were 1 hour and 1.5 hours, and variables of glucoamylase enzyme were 0.3% （w/w） and 0.5% （w/w）. To achieve these goals, the experimental work was held in laboratory scale with batch process. Firstly, tapioca solid waste was pretreated at 90 ~C and added u-amylase enzyme for 1 hour and 1.5 hours （variable of liquefaction time）. Then, substrate was cooled down to 60 ~C added with proposed variables of glucoamylase enzyme, and was analysed 24 hours after added. This experiment showed the best ratio between a-amylase and glucoamylase enzymes 0.5%：0.5% with 1 hour of liquefaction time. The highest glucose reaches 8.468% and yields 0.892 （g glucose/g starch） with starch conversion of 59.94%. KM = 0.0468 g/mL and rmax = 0.311 g/mL·h,

Chemical Characteristics and Anticoagulant Activities of Two Sulfated Polysaccharides from Enteromorpha linza (Chlorophyta)

Two sulfated polysaccharides, designated MP and SP, were extracted from the marine green alga Enteromorpha linza using hot water and then purified using ion-exchange and size-exclusion chromatography. The anticoagulant activities of MP and SP were examined by determination of their activated partial thromboplastin time （APTT）, thrombin time （TT） and prothrombin time （PT） using human plasma. Results showed that MP and SP were composed of abundant rhamnose with small amounts of xylose and glucuronic acid, whereas SP also contained a small amount of galactose. Approximate molecular weights of MP and SP were 535 and 502 kDa, respectively. As compared with SP, MP had higher contents of sulfate ester （19.0%） and uronic acid （14.9%）. The MP mainly consisted of （1→4）-linked rhamnose residues with partially sulfated groups at the C-3 position, and small amounts of （1→3, 4）-linked rhamnose, （1→2,4）-linked rhamnose, （1→4）-linked glucuronic acid and （1→4）-linked xylose residues. The SP contained abundant （1→4）-linked rhamnose with minor amounts of （1→3）-linked rhanmose, （1→3, 4）-linked rhamnose, （1→2, 4）-linked rhanmose, （1→4）4inked glucuronic acid, （1→4）-linked xylose, and （1→3）-linked galactose residues. The sulfate groups were mainly located at C-3 of （1→4）-linked rhamnose residues. Both MP and SP, in particular the former, effectively prolonged APTT and TT. This work demonstrates that MP and SP have unique structural characteristics distinct from those of other sulfated polysaccharides from Enteromorpha. The MP is a potential source of anticoagulant, and the difference in anticoagulant activities of the two sulfated polysaccharides is directly linked to the discrepancy of their chemical features.

葡萄糖母液缩合反应的原理探索及应用

以葡萄糖母液为原料,在不同葡萄糖母液浓度下,以葡萄糖淀粉酶为催化剂,研究葡萄糖母液糖组成成分的变化情况。结果表明,随着葡萄糖母液干物质浓度的提高,葡萄糖缩合为麦芽糖和异麦芽糖的反应趋势增强。同时,反应物中麦芽糖和异麦芽糖之间的比例也发生变化。反应起始,葡萄糖母液中异麦芽糖和麦芽糖的比例为0.49,异麦芽糖含量为7.16%,60℃下反应72 h后,异麦芽糖与麦芽糖比例为2.67,异麦芽糖含量增加至17.33%。葡萄糖母液经过缩合反应后,其所含的异麦芽糖含量提高了242%。

Pro-protein convertase-2/carboxypeptidase-E mediated neuropeptide processing of RGC-5 cell after in vitro ischemia

Objective To observe the change of the neuropeptide pro-protein processing system in the ischemic retina ganglion cell-5(RGC-5) cells,pro-protein convertase-2(PC2),carboxypeptidase-E(CPE) and preproneuropeptide Y(preproNPY) protein levels in the ischemic RGC-5 cells and conditioned medium were analyzed. Methods The RGC-5 cell was differentiated in 0.1 μmol/L staurosporine for 24 h and then stressed by different doses of oxygen and glucose deprivation(OGD). The acute or chronic OGD-induced cell death rates w...

Down-shifting luminescence of water soluble NaYF_4:Eu^(3+)@Ag core-shell nanocrystals for fluorescence turn-on detection of glucose

Techniques for detecting glucose are developing at a breathtaking speed because diabetes mellitus can cause many serious complications, such as blindness, high blood pressure heart disease and kidney failure. Herein, water sol- uble NaYF4：Eu^3＋@Ag core-shell nanocrystals for glucose de- tection with lower detection limit have been successfully de- veloped, using NaYF4：Eu^3＋ cores as the energy donors and Ag shells as the efficient quenchers through energy transfer. After immobilization of glucose oxidase （GOx） on the sur- face of NaYF4：Eu^3＋@Ag core-shell nanocrystals, the Ag shells can be decomposed in the presence of glucose, accompanied by down-shifting luminescence recovery. The limit of detec- tion of NaYF4：Eu^3＋@Ag was 0.12 μmol L^-1. Therefore, the NaYF4：Eu^3＋@Ag can be easily extended to the detection of a variety of H2O2-involved analytes.

Janus γ-Fe2O3/SiO2-based nanotheranostics for dual-modal imaging and enhanced synergistic cancer starvation/chemodynamic therapy

Multimodal cancer synergistic therapy exhibited remarkable advantages over monotherapy in producing an improved therapeutic efficacy. In this work, Janus-type γ-Fe2 O3/SiO2 nanoparticles(JFSNs) are conjugated with glucose oxidase(GOx) for synergistic cancer starvation/chemodynamic therapy. The γ-Fe2O3 hemisphere of JFSNs can perform photoacoustic/T2 magnetic resonance dual-modal imaging of tumors.GOx on the surface of JFSNs catalyzes the decomposition of glucose and produces H2O2 for cancer starvation therapy. Subsequently, the γ-Fe2O3 hemisphere catalyzes the disproportionation of H2O2 to generate highly reactive hydroxyl radicals in an acidic tumor microenvironment. The close distance between GOx and JFSNs ensures adequate contact between the γ-Fe2O3 hemisphere and its substrate H2O2, thus enhancing the catalytic efficiency. This synergy of glucose depletion, biotoxic H2O2 and hydroxyl radicals significantly suppresses 4 T1 mammary tumor growth with minimal adverse effects.