外源AHLs信号物质对蓝藻生长代谢的影响

 研究了外加AHLs(acyl-HSLs)信号物质对滇池铜绿微囊藻(Microcystis aeruginosa)FACHB905以及集胞藻(Synechocysti sp.)FACHB898生长和代谢的影响.结果表明:AHLs类信号物质对2种藻具有明显的生长抑制作用.在10~80μmol·L^-1浓度范围内,2种供试藻的生物量,藻蓝蛋白(phycocyanin,PC)含量都受到了一定程度的抑制,而2种藻的可溶性糖含量和Car/Chl a却有明显地提高.随AHLs浓度的增加,铜绿微囊藻藻液中超氧自由基含量明显地增加,而集胞藻藻液中超氧自由基含量随AHLs物质浓度的增加呈下降趋势,反映出两者在适应性上是有差异的.     

Third-order optical nonlinearities for lanthanum (III) complexes with 1, 10-phenanthroline components

The nonlinear refractive indexesn ₂ for the ethanol solutions of 1, 10-phenanthroline (2.0 × 10⁻³ mol · L⁻¹), [bis(l, 10-phenanthroline) lanthanum (III)] nitrate (1.0 × 10⁻³ mol · L⁻¹), and [nitrato-fluoracil-bis (1, 10_{-phenanthroline}) lanthanum (III) ] nitrate (1.0 × 10⁻³ mol · L⁻¹) are first measured, the third-order nonlinear optical susceptibilities χ(³) and 〈γ〉 are calculated, and the mechanisms of the third-order optical nonlinearities enhanced by lanthanum (III) are discussed preliminarily.     

Effect of Cl- on photosynthetic bicarbonate uptake in two cyanobacteria Microcystis aeruginosa and Synechocystis PCC6803

Photosynthetic inorganic carbon utilization was investigated in two cyanobacteria Microcystis aeruginosa and Synechocystis PCC6803 grown in standing culture. Photosynthetic rates for the two algae reached about 10 times the theoretical CO₂ supply rate at low dissolved inorganic carbon (DIC) of 100 μmol/L, and the rates were unaffected by the addition of 20 mmol/L Na⁺, indicating that the two algae possessed Na+-independent HCO^-₃ utilization for photosynthesis under low DIC. Their photo- synthetic rates at low DIC were inhibited by higher Cl^- and the degrees of inhibition were increased with the rise of CI^- concentration, and in the presence of Diphenylamine-2-carboxylate (DPC), a reported Cl^- channel inhibitor, the rates decreased by 74%－82%, implying that putative DPC-sensitive Cl^- channels participate in Na+-independent HCO₃^- uptake for photosynthesis. The experiment of intracellular 14C-DIC accumulation for photosynthesis showed that internal DIC pools decreased by about 80% with 200 μmol/L DPC and by 64%－70% with 100 mmol/L Cl^-. The experiment of chlorophyll a fluorescence quenching showed that initial rates and extents of fluorescence quenching obviously decreased with 200 μmol/L DPC or 100 mmol/L Cl^-. The two experiments gave further evidence that putative DPC-sen- sitive Cl^- channels participate in Na+-independent HCO^-₃ uptake for photosynthesis in the two algae grown in standing culture.     

Fluorescence Determination of Artemisinin Using Hemoglobin as Catalyst and Pyronine B as Substrate

0 IntroductionMaalnadria siusb atr ompajicosr .he aAltrthe pmrisoibnlienmi (n qtihneghtraoopsiucs,QHS,Fig.1) is a sesquiterpene endoperoxide isola-ted fromArtemisia annuaL., an ancient Chineseherbal medicine usedfor treatment of fever and ma-laria.Studies of the structure and activity relation-ship have shownthat endoperoxide groupis essentialfor anti malarial activity of QHS and absence of thismoiety lead to completely loss in activity of thedrug. Many techniques have been developed to de…     

Coral reef ecosystems in the South China Sea as a source of atmospheric CO<Subscript>2</Subscript> in summer

Field measurements of air-sea CO2 exchange in three coral reef areas of the South China Sea (i.e. the Yongshu Reef atoll of the Nansha Islands, southern South China Sea (SCS); Yongxing Island of Xisha Islands, north-central SCS; and Luhuitou Fringing Reef in Sanya of Hainan Island, northern SCS) during the summers of 2008 and 2009 revealed that both air and surface seawater partial pressures of CO2 (pCO2) showed regular diurnal cycles. Minimum values occurred in the evening and maximum values in the morning. Air pCO2 in each of the three study areas showed small diurnal variations, while large diurnal variations were ob-served in seawater pCO2. The diurnal variation amplitude of seawater pCO2 was ~70 μmol mol–1 at the Yongshu Reef lagoon, 420–619 μmol mol–1 on the Yongxing Island reef flat, and 264–579 μmol mol–1 on the reef flat of the Luhuitou Fringing Reef, and 324–492 μmol mol–1 in an adjacent area just outside of this fringing reef. With respect to spatial relations, there were large differences in air-sea CO2 flux across the South China Sea (e.g. ~0.4 mmol CO2 m–2 d–1 at Yongshu Reef, ~4.7 mmol CO2 m–2 d–1 at Yongxing Island, and ~9.8 mmol CO2 m–2 d–1 at Luhuitou Fringing Reef). However, these positive values suggest that coral reef ecosystems of the SCS may be a net source of CO2 to the atmosphere. Additional analyses indicated that diurnal variations of surface seawater pCO2 in the shallow water reef flat are controlled mainly by biological metabolic processes, while those of deeper water lagoons and outer reef areas are regulated by both biological metabolism and hydrodynamic factors. Unlike the open ocean, inorganic metabolism plays a significant role in influencing seawater pCO2 variations in coral reef ecosystems.     

Significance of native arsenic in the Baogutu gold deposit, western Junggar, Xinjiang, NW China

Native arsenic together with comb quartz and stibnite is found in the Baogutu gold deposit, western Junggar (Xinjiang), NW China. It is anhedral with various grain size (<0.001 to 2 mm), and contains 98 wt% to 98.7 wt% As. Micro-granular electrum, the main auriferous mineral in the Baogutu gold deposit, is commonly enclosed in or closely accompanied by native arsenic. Three ore-forming paragenetic stages could be identified. Native arsenic mainly formed at stage II which is also the major stage for gold deposition. Mineral assemblage formed at this stage is native arsenic-stibnite-electrum-arseno- pyrite-miargyrite-freibergite-pyrrhotite-pyrite. Based on native arsenic and its coexisting minerals, the temperature (230 to 170℃), oxygen fugacity (logfO₂ = −42―−56.5) and sulfur fugacity (logf S₂ = −13.3― −16.6) of stage II are estimated. From stage I to stage II, the temperature, sulfur fugacity and S²⁻ con-centration of hydrothermal fluid decrease obviously, whereas the As concentration increases. Coexis-tence with native arsenic of electrum and its contents of 0.5 wt%―1.3 wt% As suggest that As is im-portant to transport Au when S²⁻ concentration decrease in hydrothermal fluid. Crystallization of native arsenic induced the deposition of electrum and consequently the formation of the Baogutu gold deposit.     

Phytosterol of <Emphasis Type="Italic">Potamogeton maackianus</Emphasis> and its change under the stress of <Emphasis Type="Italic">Microcystis aeruginosa</Emphasis>

The phytosterol in Potamogeton maackianus was identified and quantified. From the hexane extracts analysis of unsaponifiable fraction of P. maackianus by gas chromatography-mass spectrometry(GC-MS), campesterol, stigmasterol and β-sitosterol were the main phytosterols in P. maackianus with the contents of 0.368, 1.17 and 0.824 mg·g⁻¹, respectively. After 7 days’ exposure under M. aeruginosa, the contents of campesterol, stigmasterol and β-sitosterol in P.maackianus increased by 23.1%, 40.4% and 40.8%, respectively. Both the contents of 24-ethyl sterols (sitosterol and stigmasterol) and 24-methyl sterol (campesterol) in macrophytes increased, but the percentage of total sterol composition did not change significantly. This result indicated that the content of 4-demethyl sterols and the second alkylation of the phytosterol side-chain at C-24 in P. maackianus were affected by M. aeruginosa. Biography: ZHANG Shenghua(1979–), female, Ph.D. candidate, research direction: environmental biology.     

The comparative investigation on redox property and second-order nonlinear response of Keggin-type α-[PM12O39NPh]3− (M = W and Mo) and Mo6NPh

Keggin-type phenylimido-polyoxometalates α-[PM₁₂O₃₉NPh]³⁻ (M = W and Mo) have been systematically investigated on the electronic structures, redox as well as nonlinear optical (NLO) properties by density functional theory (DFT). The strong M≡N bond confirmed by natural bond orbital (NBO) analysis comprises one s bond and two π bonds, the same as Mo≡N in [Mo₆O₁₈NPh]²⁻. Furthermore, phenylimido segment effectively modifies the electronic properties of α-[PM₁₂O₃₉NPh]³⁻. On one hand, when enlarging the inorganic cluster from {Mo₆O₁₈} to {PMo₁₂O₃₉}, the energy gap between HOMO and LUMO in α-[PMo₁₂O₃₉NPh]³⁻ decreased, resulting in enormously anodic shift for the reduction potential, while the excitation energy is less and the total second-order polarizability β ₀ is up to 438-3×10⁻³⁰ esu, which is nearly 10 times larger than that of [Mo₆O₁₈NPh]²⁻. On the other hand, when metal W in α-[PM₁₂O₃₉NPh]³⁻ is substituted by Mo, the interaction between Mo and N is enhanced and the redox ability becomes stronger. The β ₀ value for α-[PMo₁₂O₃₉NPh]³⁻ is more than 5 times higher than that of α-[PW₁₂O₃₉NPh]³⁻. It indicates that changing appropriate metal or enlarging the inorganic cluster will improve the redox properties and second-order nonlinear response. Moreover, the electron transition for three compounds mentioned above occurred mainly from organoimido segment (as the electron donor) to polyanion cluster (as the acceptor). As a result, α-[PMo₁₂O₃₉NPh]³⁻ may be a promising candidate for oxidant and nonlinear optical material. Electronic Supplementary Material  Supplementary material is available for this article at and is accessible for authorized users. Supported by National Natural Science Foundation of China (Grant No. 20573016), Training Fund of NENU’s Scientific Innovation Project (Grant No. NENUSTC07017), Science Foundation for Young Teachers of Northeast Normal University (Grant No. 20070304) and Program for Changjiang Scholars and Innovative Research Team in University (PCSIRT)     

Seasonal variation and sources of low molecular weight organic acids in precipitation in the rural area of Anshun

Low molecular weight (LMW) organic acids are important and ubiquitous chemical constituents in the atmosphere. A comprehensive study of the chemical composition of precipitation was carried out from June 2007 to June 2008 at a rural site in Anshun, in the west of Guizhou Province, China. During this period, 118 rainwater samples were collected and the main LMW carboxylic acids were determined using ion chromatography. The average pH of rainwater was 4.89 which is a typical acidic value. The most abundant carboxylic acids were formic acid (volume weight mean concentration: 8.77 μmol L⁻¹) and acetic acid (6.90 μmol L⁻¹), followed by oxalic acid (2.05 μmol L⁻¹). The seasonal variation of concentrations and wet deposition fluxes of organic acids indicated that direct vegetation emissions were the main sources of the organic acids. Highest concentrations were observed in winter and were ascribed to the low winter rainfall and the contribution of other air pollution sources northeast of the study area. The ratio of formic and acetic acids in the precipitation ([F/A] _T ) was proposed as an indicator of pollution source. This suggested that the pollution resulted from direct emissions from natural or anthropogenic sources. Comparison with acid precipitation in other urban and rural areas in Guizhou showed that there was a decreasing contribution of LMW organic acids to free acidity and all anions in rainwater from urban to remote rural areas. Consequently, it is necessary to control emissions of organic acids to reduce the frequency of acid rain, especially in rural and remote areas.     

Synthesis and surface activity of biquaternary ammonium salt gemini surfactants with ester bond

The synthesis and surface activity of gemini surfactants with ester bond as spacer are described. Their critical micelle concentrations （CMC） are much lower than that of conventional surfactants, i.e., 1.62×10^-5, 1.38×10^-5, 1.28×10^-5 mol·L^-1 for gemini Ⅰ, Ⅱ and Ⅲ respectively. Surface tension at the CMC of gemini Ⅰ, Ⅱ and Ⅲ were 36.4, 38.5, 41.2 mN · m^-1. The physico-chemical properties such as Krafft points, foaming abilities and emulsifying power were also investigated. It is found that the title compounds have low Krafft points and show good solubility in water. The gemini surfactants synthesized also exhibit good foaming properties and excellent emulsifying power toward toluene. The foaming abilities and emulsifying power increase with the increase in carbon number of hydrophobic chain, and this might be caused by the sheer viscosity of different gemini surfactant solution.     

Fluorescence determination of tetracycline by Eu (III)/TOPO/TC system

The fluorescence intensity of the europium (III)-tetracycline (TC) complex can be greatly increased by TOPO, TBP, TTA and surfactans, among which TOPO showed the greatest enhancement. The formation of ternary complex was proposed to be responsible for the fluorescence enhancement. The Eu (III)-TOPO-TC system can be applied to the determination of trace amounts of tetracycline in urinous sample; the maximum fluorescence intensity was obtained in the pH range of 8.2∼9.5 with excitation and emission wave-lengths at 396 nm and 616 nm, respectively. The linear range for tetracycline is 6.0×10⁻⁹−5.0×10⁻⁷ mol·L⁻¹. The detection limit is 2.0×10⁻⁹ mol·L⁻¹(SNR=3). Experimental conditions and the mechanism of the enhancement effect are discussed. Supported by National Natural Science Foundation of China Huang Chunbao: born in 1958, Associate professor     

An acetylcholinesterase inhibitor, 3-benzidino-5-methyl-6-phenylpyridazine, blocking outward potassium currents in acutely isolated rat hippocampal pyramidal neurons

3-benzidino-5-methyl-6-phenylpyridazine (BMP) inhibited electric eel acetylcholinesterase (AChE), with IC50 being 0.58 μmol·L-1. As an AChE inhibitor, the effects of BMP on delayed rectifier potassium current (IK(DR)) and transient outward potassium current (IK(A)) in acutely isolated rat hippocampal pyramidal neurons were studied using the whole cell patch-clamp technique. BMP (0.1―50 μmol·L-1) inhibited IK(DR) and IK(A) in a concentration-dependent and voltage-independent manner. The IC50 value for the bl...     

Organic media enhanced spectrofluorimetric determination of L-ascorbic acid

A novel spectrofluorimetric method for the determination of L-ascorbic acid is proposed. It is based on the inhibition of L-ascorbic acid on the formation of 2,3-diaminophenazine, which is an oxidation product ofo-phenylenediamine catalyzed by laccase. The fluorescence (at λ_cx/λ_cm=464 nm /530 nnm) was enhanced strongly in the presence of organic media. The mechanism ofo-phenylenediamine oxidation reaction catalyzed by laccase in the presence of L-ascorbic acid is discussed. L-ascorbic acid is determined in the ethanol, 1,4-dioxane and acetone over the linear range of 4.0×10⁻⁷≈1.2×10⁻⁴ mol/L, 4.0×10⁻⁷≈8.0×10⁻⁵ mol/L and 4.0×10⁻⁷≈1.0×10⁻⁴ mol/L with a detection limit of 1.20×10⁻⁵ mol/L, 1.19×10⁻⁵ mol/L and 1.24×10⁻⁸ mol/L, respectively. The method has been successfully applied to the simple and rapid determination of L-ascorbic acid in pharmaceuticals and milk powder. Supported by National Natural Science Foundation of China and the Special Funds of State Education Committe for Doctorate Research Huang Zuyun: born in Aug. 1963. Ph.D     

Adrenomedullin inhibits the endothelin production induced by urotensin II in rat vascular smooth muscle cells

The aim of this study was to observe the effects of adrenomedullin (ADM) on endothelin (ET) production induced by urotensin Ⅱ (UⅡ) in rat vascular smooth muscle cells (VSMCs). Cultured VSMCs which were incubated with UⅡ (10^-8 mol/L) and with various concentrations of ADM were used to measure the VSMCs ³H-TdR incorpora- tion, the activity of extracellular signal-regulated kinase (ERK), the amount of ET mRNA and ET production in VSMCs. In this work we found that incubation with UⅡ(10^-8 mol/L) increased obviously the amount of ET mRNA in VSMCs and ET production in medium, however, coincubation with ADM (10^-10—10^-8 mol/L) and UⅡ(10^-8 mol/L) reduced the amount of ET mRNA by 15%, 24% and 45% (P< 0.01) respectively, compared with UⅡ alone. The content of ET in medium was 14.13, 11.38 and 11.00 pg/mL. ADM alone (10^-8 mol/L) had no effect on ET production in VSMCs. UⅡ (10^-8 mol/L) promoted the 3H-TdR incorpo- ration and activity of ERK in VSMCs. ADM inhibited VSMCs 3H-TdR incorporation and activation of ERK in a concentration-dependent manner. Compared with UⅡ group, after coincubation with ADM (10^-10—10^-8 mol/L) and UⅡ (10^-8 mol/L) the VSMCs 3H-TdR incorporation was decreased by 7% (P > 0.05), 32% (P < 0.05) and 41% (P < 0.01), respectively, and the activity of ERK was decreased by 24% (P > 0.05), 32% (P < 0.05) and 36% (P < 0.05), re- spectively, in a concentration-dependent manner. The results show that in cultured VSMCs ADM inhibits ET mRNA expression, ET production and proliferation stimulated by UⅡ, and that inhibitory effect of ADM on UⅡ bioaction could be mediated through inhibiting MAPK pathway.     

Microcalorimetric study on the antibiotic activity of clarithromycin and erythromycin

By using LKB-2277 Bioactivity Monitoring System, the heat effect changes in the process of inhibitory action of clarithromycin and erythromycin onEscherichia coli at 37°C were determined. Quantitative analysis showed that relationship between antibiotic concentrationc and rate contantk ofEscherichia coli growth, and half inhibitory ratio concentration IC₅₀: clarithromycin:k=0. 030 03–1. 1736×10⁻³ c, 8. 45 mg ·L⁻¹; erythromycin:k=0.031 08–8.4657×10⁻⁴ c, 14. 45 mg·L⁻¹. As a result of the microcalorimetry experiments, it not only indicated that antibacterial activity of clarithromycin was stronger than that of erythromycin, but also reported the changeable features of thermodynamics of the bacterial cell in biological, biochemical and metabolic process under different drug action. Foundation item: Supported by Natinal Natural Science Fundation of China (29973030), Natural Science Fundation of Hubei Province (98J052) and Post-doctoral Science Fundation of China Biography, SHEN Xue-song (1956-), Associate professor Research direction: biothermochemistry.     

Effect of CO<Subscript>2</Subscript> concentrations on the activity of photosynthetic CO<Subscript>2</Subscript> fixation and extracelluar carbonic anhydrase in the marine diatom <Emphasis Type="Italic">Skeletonema costatum</Emphasis>

The growth and activity of photosynthetic CO2 uptake and extracellular carbonic anhydrase (CAext) of the marine diatom Skeletonema costatum were investigated while cultured at different levels of CO2 in order to see its physio-logical response to different CO2 concentrations under either a low (30 靘ol·m-2·s-1) or high (210 靘ol·m-2·s-1) irradiance. The changes in CO2 concentrations (4—31 靘ol/L) affected the growth and net photosynthesis to a greater extent under the low than under the high light re-gime. CAext was detected in the cells grown at 4 mol/L CO2 but not at 31 and 12 靘ol/L CO2, with its activity being about 2.5-fold higher at the high than at the low irradiance. Photo- synthetic CO2 affinity (1/ K1/2(CO2)) of the cells de-creased with increased CO2 concentrations in culture. The cells cultured under the high-light show significantly higher photosynthetic CO2 affinity than those grown at the low-light level. It is concluded that the regulations of CAext activity and photosynthetic CO2 affinity are dependent not only on CO2 concentration but also on light availability, and that the de-velopment of higher CAext activity and CO2 affinity under higher light level could sufficiently support the photosyn-thetic demand for CO2 even at low level of CO2.     

Electrochemical determination of trace amounts of lead (II) and cadmium (II) at a calix[6]arene modified carbon paste electrode

An electrochemical method for the simultaneous determination of lead(II) and Cadmium(II) with a calix[6]arene modified carbon paste electrode (CPE) has been developed. Pb²⁺ and Cd²⁺ were accumulated at the surface of the modified electrode via formation of chemical complexes with calix[6]arene, and reduced at 1.40 V. During the following anodic potential sweep, reduced lead and cadmium were oxidized, and two well-defined striping peaks appeared at about −0.60 V and −0.84 V. Compared with a bare carbon paste electrode, the calix[6]arene modified CPE greatly improves the sensitivity of determining lead and cadmium. The stripping peak currents change linearly with the concentration of Pb²⁺ 3.0×10⁻⁸–8.0×10⁻⁶ mol·L⁻¹ and with that of Cd²⁺ 6.0×10⁻⁸–1.0×10⁻⁵ mol ·L⁻¹. The detection limits of Pb²⁺ and Cd²⁺ are found to be 8.0×10⁻⁹ mol·L⁻¹ and 2.0×10⁻⁸ mol·L⁻¹, respectively. The modified carbon paste electrode was applied to determine trace levels of lead and cadmium in water samples. Comparing with that of atomic absorption spectrometry, the results suggests that the calix[6]arene modified CPE has great potential for the practical sample analysis. Foundation item: Supported by the National Natural Science Foundation of China (60171023) Biography: JI Xiao-bo (1980-), male, Master, research direction: electroanalytical chemistry.     

Comparative analysis of the dissolution kinetics of galena in binary solutions of HCl/FeCl<Subscript>3</Subscript> and HCl/H<Subscript>2</Subscript>O<Subscript>2</Subscript>

A comparative study of the dissolution kinetics of galena ore in binary solutions of FeCl₃/HCl and H₂O₂/HCl has been undertaken. The dissolution kinetics of the galena was found to depend on leachant concentration, reaction temperature, stirring speed, solid-to-liquid ratio, and particle diameter. The dissolution rate of galena ore increases with the increase of leachant concentration, reaction temperature, and stirring speed, while it decreases with the increase of solid-to-liquid ratio and particle diameter. The activation energy (E _a) of 26.5 kJ/mol was obtained for galena ore dissolution in 0.3 M FeCl₃/8.06 M HCl, and it suggests the surface diffusion model for the leaching reaction, while the E _a value of 40.6 kJ/mol was obtained for its dissolution in 8.06 M H₂O₂/8.06 M HCl, which suggests the surface chemical reaction model for the leaching reaction. Furthermore, the linear relationship between rate constants and the reciprocal of particle radius supports the fact that dissolution is controlled by the surface reaction in the two cases. Finally, the rate of reaction based on the reaction-controlled process has been described by a semiempirical mathematical model. The Arrhenius and reaction constants of 11.023 s⁻¹, 1.25×10⁴ and 3.65×10² s⁻¹, 8.02×10⁶ were calculated for the 0.3 M FeCl₃/8.06 M HCl and 8.06 M H₂O₂/8.06 M HCl binary solutions, respectively.     

Electrocatalytic oxidation of H<Subscript>2</Subscript>O<Subscript>2</Subscript> at a carbon paste electrode modified with a nickel (II)-5, 11, 17, 23-tetra-tert-butyl-25, 27-bis(Diethylcarbamoylmethoxy) calix[4] arene complex and its application

Electrochemical behavior of a carbon paste electrode (CPE) modified with nickel(II)-5, 11, 17, 23-tetra-tert-butyl-25, 27-bis(diethylcarbamoylmethoxy) calix[4]arene (Ni (II)-L) complex and its electrocatalytic activity towards the oxidation of hydrogen peroxide were investigated by cyclic voltammetric technique in a 5.0×10⁻² mol/L NaClO₄+1.0×10⁻³ mol/L NaOH solution. It was found that Ni(II)-L acts as an effective catalyst for the oxidation of hydrogen peroxide. The modified electrode exhibited a linear response over a hydrogen peroxide concentrations in the range of 2.0×10⁻⁵−1.0×10⁻⁴ mol/L with a detection limit as low as 1.0×10⁻⁶ mol/L. The relative standard deviation was 3.5% for 5 successive determinations of H₂O₂ at 1.0×10⁻⁵ mol/L. The modified electrode was used successfully in rainwater analysis. Foundation item: Supported by the Natural Science Foundation of Hubei Province (98J040). Biography: Li Chun ya(1972-), Ph. D. candidate, research direction: electroanalysis and electrosynthesis.     

Direct electrochemistry and electrocatalysis of horseradish peroxidase in MnO2 nanosheet film

A novel material MnO2 nanosheet has been used as the support matrix for the immobilization of horseradish peroxidase （HRP）. HRP entrapped in MnO2 nanosheet film exhibits facile direct electron transfer with the electron transfer rate constant of 6.86 s^-1. The HRP/MnO2 nanosheet film gives a reversible redox couple with the apparent formal peak potential （E^0＇） of -0.315 V （vs. Ag/AgCl） in pH 6.5 phosphate buffer solution （PBS）. The formal potential E^0＇ of HRP shifts linearly with pH with a slope of -53.75 mV.pH^-1, denoting that an electron transfer accompanies single-proton transportation. The immobilized HRP shows an electrocatslytic activity to the reduction of H2O2. The response time of the biosensor for H2O2 is less than 3 s, and the detection limit is 0.21 μmol · L^-1 based on signal/noise = 3.