Trypsin immobilization by direct adsorption on metal ion chelated macroporous chitosan-silica gel beads

Silica gel bead coated with macroporous chitosan layer (CTS-SiO₂) was prepared, and the metal immobilized affinity chromatographic (IMAC) adsorbents could be obtained by chelating Cu²⁺, Zn²⁺, Ni²⁺ ions, respectively on CTS-SiO₂, and trypsin could be adsorbed on the IMAC adsorbent through metal–protein interaction forces. Batch adsorption experiments show that adsorption capacity for trypsin on these IMAC adsorbent variated with change of pH. The maximal adsorption reached when the solution was in near neutral pH in all three IMAC adsorbents. Adsorption isothermal curve indicated that maximal adsorption capacity could be found in the Cu²⁺-CTS-SiO₂ with the value of 4980 ± 125 IU g⁻¹ of the adsorbent, while the maximal adsorption capacity for trypsin on Zn²⁺ and Ni²⁺ loaded adsorbent was 3762 ± 68 IU g⁻¹ and 2636 ± 53 IU g⁻¹, respectively. Trypsin immobilized on the IMAC beads could not be desorbed by water, buffer and salt solution if the pH was kept in the range of 5–10, and could be easily desorbed from the IMAC beads by acidic solution and metal chelating species such as EDTA and imidazole. The effect of chelated metal ions species on CTS-SiO₂ beads on the activity and stability of immobilized trypsin was also evaluated and discussed. Trypsin adsorbed on Zn-IMAC beads retained highest amount of activity, about 78% of total activity could be retained. Although the Cu-IMAC showed highest affinity for trypsin, only 25.4% of the calculated activity was found on the beads, while the activity recovery found on Ni-IMAC beads was about 37.1%. A remarkable difference on stability of trypsin immobilized on three kinds of metal ion chelated beads during storage period was also found. Activity of trypsin on Cu-IMAC decreased to 24% of its initial activity after 1-week storage at 4 °C, while about 80% activity was retained on both Ni-IMAC and Zn-IMAC beads. Trypsin immobilized on Zn-CTS-SiO₂ could effectively digest BSA revealed by HPLC peptide mapping.     

一株高效解磷真菌新菌株的筛选鉴定及解磷特性

从辽宁省辽中县多年耕种的日光温室番茄根际土壤中筛选出一株解磷真菌,通过菌落形态特征和ITS rDNA序列对比,鉴定该菌株为草酸青霉菌的一株新菌株,将其命名为PSF1.该菌株能利用葡萄糖、蔗糖、乳糖、半乳糖、可溶性淀粉等多种碳源和硫酸铵、氯化铵、硝酸铵、硝酸钾、尿素等多种氮源进行生长代谢并表现出较强的解磷能力,在C/N 10∶1~60∶1、初始pH 7~8的条件下生长情况较好且解磷能力较高.该菌株有很强的产酸能力,在培养过程中培养液pH由7.00~7.50下降至2.06~4.87;在4种磷源培养液中的最高解磷量分别为磷酸三钙(869.62 mg·L^-1)>磷矿粉(233.56 mg·L^-1)>磷酸铝(44.77 mg·L^-1)>磷酸铁(28.42 mg·L^-1).Pearson相关分析表明,菌株在磷酸三钙、磷矿粉和磷酸铁培养液中的解磷量与pH的变化之间呈极显著负相关关系,在磷酸铝培养液中无显著相关关系.菌株PSF1解磷能力强,生长条件广,推测其在土壤中有较强的解磷能力.     

黑曲霉解磷能力的影响因素及培养条件优化

本文探讨了培养条件下黑曲霉解磷能力的主要影响因素。通过单孢株筛选得到解磷能力较强的菌株Xj-2,其在液体培养基中的解磷能力达到539.90 mg·L^-1。解磷发酵动力学模型模拟发现,其解磷能力在培养的第4 天达到平稳,可作为终止发酵时间。不同磷源培养液中菌株Xj-2的解磷量依次为磷酸钙(539.90 mg·L^-1)>磷酸锌(238.45 mg·L^-1)>磷酸铁(182.64 mg·L^-1)>磷矿粉(71.80 mg·L^-1)>磷酸铝(24.40 mg·L^-1)。通过单因素试验并结合响应面优化,研究了其解磷最佳条件。结果表明: 碳源对Xj-2的解磷能力影响最大,其次是菌群密度和培养液pH。当培养温度35 ℃、转速160 r·min^-1、培养液pH 6.0、氮源(尿素)浓度0.79 g·L^-1、碳源(葡萄糖)浓度10.00 g·L^-1、菌群密度3.8%、培养时间为4 d时,Xj-2的解磷能力最高,为616.81 mg·L^-1。     

石油焦基高比表面积活性炭对废水中CODCr的吸附能力

探讨了高比表面积活性炭(HSAAC)吸附水中COD_Cr时,活性炭用量、pH值和吸附时间等因素对COD_Cr吸附量和去除率的影响。实验结果表明,HSAAC用量越大,去除COD_Cr效果越好。当HSAAC用量为2.0g·L^-1,pH=3时,去除率达到78%以上;在酸性条件下HSAAC对COD_Cr的去除效果较好;HSAAC对废水中COD_Cr的吸附发生在前30min;COD_Cr浓度低于60mg·L^-1时,处理后COD_Cr的残余质量浓度低于地表水环境质量Ⅰ类标准(15mg·L^-1)。用碱再生HSAAC,一次再生率达94.22%,二次再生率达到了86.90%。说明高比表面积活性炭在适宜条件下对COD_Cr具有较好的吸附性能和良好的再生效果。     

Removal of lead (Pb(2+)) by the cyanobacterium Gloeocapsa sp

Pb²⁺ removal ability of the viable-freshwater cyanobacterium Gloeocapsa sp. was studied in batch experiments. Gloeocapsa sp. was cultured in the Medium 18 with pH adjusted to 3, 4, 5, 6 and 7. Growth was subsequently determined based on the increase of chlorophyll-a content. Gloeocapsa sp. was able to grow at all pH levels tested, except at pH 3. Removal of Pb²⁺ was then further studied under pH 4. The results showed that Pb²⁺ concentration in the range of 0–20 mg L⁻¹ was not inhibitory to Gloeocapsa sp. growth but reduced its Pb²⁺ removal efficiency (by 4.5% when Pb²⁺ concentration increased from 2.5 to 20 mg L⁻¹). Pb²⁺ removal characteristics followed the Langmuir adsorption isotherm with the maximum removal capacity (q_max) of 232.56 mg g⁻¹. Adsorption of Pb²⁺ by this cyanobacterium followed the second order rate reaction and intraparticle diffusion was likely the rate-determining step. The initial rate of Pb²⁺adsorption during intraparticle diffusion was slower under light than under dark conditions, indicating that light probably slowed down the initial rate of intraparticle diffusion through the repulsion effects on cell membrane.     

不同炭化温度制备的蚕丝被废弃物生物炭对重金属Cd2+的吸附性能

以蚕丝被废弃物为原料,在300、500和700 ℃高温缺氧条件下热解炭化制备成3种生物炭(BC300、BC500和BC700).利用扫描电镜(SEM)、傅里叶红外光谱仪(FT-IR)、X-射线衍射仪(XRD)、比表面积分析仪等对其理化性质进行表征,并研究了不同温度下制备的生物炭对溶液中Cd²⁺的吸附特性.结果表明: 随着炭化温度上升,BET比表面积、pH、灰分均增大,生物炭表面形态结构越来越不规则.XRD结果显示:不同温度下获得的生物炭中均含有一定量的方解石,FT-IR光谱图上的峰主要为-OH和方解石典型的吸收峰;pH对生物炭吸附Cd²⁺的影响不大;Langmuir方程能更好地拟合3种生物炭对Cd²⁺的吸附等温过程,其最大吸附量分别为25.61、52.41和91.07 mg·g^-1.3种生物炭对Cd²⁺吸附过程均更符合准二级动力学方程,且BC700对Cd²⁺的吸附效果最佳.进一步研究离子浓度及阳离子共存对BC700吸附Cd²⁺的影响,结果显示: NaCl浓度越高,对Cd²⁺的吸附抑制越明显;共存阳离子中,Ca²⁺和Mg²⁺对Cd²⁺的吸附抑制更明显,而K⁺几乎无影响.因此,以蚕丝被废弃物制备的生物炭作为去除水体中Cd²⁺的吸附剂具有较强的应用潜力.     

The adsorption of copper(II) ions on to dehydrated wheat bran (DWB): determination of the equilibrium and thermodynamic parameters

The adsorption of copper(II) ions on to dehydrated wheat bran (DWB), a by-product of the flour process, was investigated as a function of initial pH, temperature, initial metal ion concentration and adsorbent dosage. The optimum adsorption conditions were initial pH 5.0, initial copper concentration 100 mg l⁻¹, temperature 60 °C and adsorbent dosage 0.1 g. The adsorption equilibrium was described well by the Langmuir isotherm model with maximum adsorption capacity of 51.5 mg g⁻¹ of copper(II) ions on DWB. The observation of an increase in adsorption with increasing temperature leads to the result that the adsorption of copper(II) ions on DWB is endothermic in nature. The thermodynamic parameters such as enthalpy, free energy and entropy changes were calculated and these values show that the copper(II)-DWB adsorption process was favoured at high temperatures.     

改性粉煤灰对废水中镉的吸附作用

以工业固废粉煤灰(FA)为原料,将其与NaOH和Ca(OH)₂混合后,在250 ℃下焙烧1.5 h,制得改性粉煤灰(MFA).利用扫描电镜(SEM)、能量色散X射线光谱仪(EDS)、X-射线衍射仪(XRD)、全自动比表面积分析仪(BET)和傅里叶红外光谱仪(FTIR)等技术分析MFA的理化性质.BET分析表明,MFA的比表面积比FA增大了20.6倍.SEM分析表明,FA表面玻璃相溶解,表面粗糙,具有多孔结构.FTIR分析表明,-OH官能团在Cd²⁺的吸附过程中具有重要作用.静态吸附试验研究表明,当MFA的投加量为0.2 g、Cd²⁺初始浓度为100 mg·L^-1、溶液pH为7、25 ℃下吸附90 min时,Cd²⁺的去除率达到97.3%;共存阳离子(K⁺、Na⁺、Mg²⁺和Ca²⁺)均会抑制MFA对Cd²⁺的吸附,其中Ca²⁺的抑制作用最强.Langmuir等温吸附模型和准二级吸附动力学方程能较好地描述Cd²⁺在MFA上的吸附行为,且最大吸附量为55.77 mg·g^-1.热力学研究表明,MFA对Cd²⁺的吸附是一个自发的吸热过程.MFA吸附能力优于FA,在废水处理方面有一定的应用前景.     

Survival and infectivity of a Rhizobium meliloti strain maintained in water and buffer suspensions

Rhizobium meliloti B323 cells were suspended in deionized water, phosphate buffer pH 6.5 and 5.5 and these buffers supplemented with Ca²⁺, Mg²⁺ (1 mmol/l) and Fe³⁺ (0.1 mmol/l). Initial cell count was 1.10⁸ cells/ml. The viable count of rhizobia suspended in buffer at pH 6.5, with and without salts, remained constant or even increased during storage. Cells suspended in buffer at pH 5.5 with salts, decreased in numbers in the first 5 months, then, until the 10th month, the count remained at 10⁵ cells/ml. Rhizobia suspended in buffer at pH 5.5 and deionized water decreased in viability almost to zero by the 10th month. In those suspensions where viability was maintained, the symbiotic infectivity of cells was also maintained as compared with a control performed with fresh cultured rhizobia. In suspensions in deionized water and buffer at pH 5.5 where the viability diminished during the experiment, the rhizobia lost their ability to infect roots immediately after inoculation but maintained their capacity to form late nodules on the hosts.     

Red blood cell adrenergic responses in Amazonian teleosts

Erythrocytes from Amazonian teleosts of the Rio Negro were surveyed for the presence of adrenergically mediated Na⁺/H⁺ exchange. Washed red blood cells (RBCs) incubated in HEPES-buffered Cortland saline were stimulated with 10⁻⁴ M L-adrenaline. The adrenergic response was clearly present in two characids, the tambaqui Colossoma macropomom and the jaraqui Semaprochilodus insignis , as demonstrated by a decrease in the pH_c-pH_i gradient across the RBC membrane, an uptake of Na⁺ from the extracellular medium, and RBC swelling. The latter was signalled by increased mean corpuscular volume (MCV) and decreased mean corpuscular haemoglobin concentration (MCHC). The response did not occur in two other characids, the black piranha Serrasalmus rhombeus and the aracu Leporinus fasciatus or in two silurid catfish, the piranambu Pinirampus pirinampu and the acari-bodo, armoured catfish, Pterygoplichthys multiradiatus . In acari-bodo, the Na⁺/H⁺ exchange response was similarly lacking under anoxic conditions. Oxygenated/deoxygenated comparisons revealed the presence of a marked Root effect in jaraqui and its absence in acari-bodo. GTP dominated over ATP as the major intracellular phosphate in all six species. There were no significant changes in any nucleoside phosphate (ATP, ADP, AMP, GTP, GDP, or GMP) in response to adrenaline in any species.     

Biosorption of Lead from Industrial Wastewater Using Chrysophyllum albidum Seed Shell

The shell of the seed of Chrysophyllum albidum carbon was used to adsorb lead (Pb) from aqueous solution, the sorption process with respect to its equilibria and kinetics as well as the effects of pH, contact time, adsorbent mass, adsorbate concentration, and particle size on adsorption were also studied. The most effective pH range was found to be between 4.5 and 5 for the sorption of the metal ion. The first-order rate equation by Lagergren was tested on the kinetic data and the adsorption process followed first-order rate kinetics. Isotherm data were analyzed for possible agreement with the Langmuir and Freundlich adsorption isotherms; the Freundlich and Langmuir models for dynamics of metal ion uptake proposed in this work fitted the experimental data reasonably well. However, equilibrium sorption data were better represented by Langmuir model than Freundlich. The adsorption capacity calculated from Langmuir isotherm was 72.1 mg Pb (II) g^{- 1} at initial pH of 5.0 at 30°C for the particle size of 1.00 to 1.25 mm with the use of 2.0 g/100 ml adsorbent mass. The structural features of the adsorbent were characterized by Fourier transform infrared (FTIR) spectrometry; the presence of hydroxyl, carbonyl, amide, and phosphate groups confirms the potential mechanism adsorption of the adsorbent. This readily available adsorbent is efficient in the uptake of Pb (II) ion in aqueous solution, thus, it could be an excellent alternative for the removal of heavy metals and organic matter from water and wastewater.     

Removal and recovery of phosphorus from water by means of adsorption onto orange waste gel loaded with zirconium

Orange waste, an available biomass, was immobilized with zirconium(IV) to investigate its feasibility for phosphate removal from an aquatic environment. Kinetics, effects of pH and foreign anions, and the adsorption isotherm for phosphate have been examined. The adsorption capacity has been compared to that of two commercially available adsorbents such as zirconium ferrite and MUROMAC XMC 3614. The prepared gel was an effective adsorption gel for phosphate removal with a reasonably high sorption capacity of 57mg-P/g, which was four times higher than that of zirconium ferrite. The highest removal of phosphate was observed at low pH, whereas higher pH suppressed phosphate removal, but even up to pH 9 more than 85% phosphate removal was observed. Adsorbed phosphate was eluted by NaOH solution. Fixed bed column-mode experiments confirmed the complete adsorption of phosphate in continuous-mode operation. Throughout the operating conditions, zirconium was not leaked.     

解脂假丝酵母(Candida lipolytica)对铜的吸附

研究了解脂假丝酵母的表面特性及培养时间、pH值、铜浓度、菌体投加量、吸附时间等因素对铜吸附的影响,并探讨了吸附动力学特征。结果表明,菌体表面可能有-OH和-PO₄^3-,培养96 h的菌体吸附性能最佳,适宜pH为4.0-6.0,适宜菌体投加量为25.0g·L^-1(湿重)。在初始浓度为20mg·L^-1的铜溶液中投加25.0g·L^-1(湿重)的菌体,吸附2h,铜的去除率最高达86.5%。铜浓度为5,10mg·L^-1时,铜的去除率高达95%以上。动力学分析表明,在实验设定的浓度范围内解脂假丝酵母对铜的吸附基本符合Freundlich吸附模型。红外光谱分析表明吸附后-OH吸收峰蓝移18cm^-1,其它吸收峰没有明显的变化。     

Factors affecting the protease activity of venom from jellyfish Rhopilema esculentum Kishinouye

In this paper, the effects of some chemical and physical factors such as temperature, pH values, glycerol, and divalent metal cations on the protease activity of venom from jellyfish, Rhopilema esculentum Kishinouye, were assayed. Protease activity was dependent on temperature and pH values. Zn²⁺, Mg²⁺, and Mn²⁺ in sodium phosphate buffer (0.02 M, pH 8.0) could increase protease activity. Mn²⁺ had the best effects among the three metal cations and the effect was about 20 times of that of Zn²⁺ or Mg²⁺ and its maximal protease activity was 2.3 × 10⁵ U/mL. EDTA could increase protease activity. PMSF had hardly affected protease activity. O-Phenanthroline and glycerol played an important part in inhibiting protease activity and their maximal inhibiting rates were 87.5% and 82.1%, respectively.     

Interfacial tension of phosphatidylcholine-phosphatidylserine system in bilayer lipid membrane

The effect of pH of electrolyte solution on the interfacial tension of lipid membrane formed of phosphatidylcholine (PC, lecithin)–phosphatidylserine (PS) system was studied. In this article, three models describing the H⁺ and OH⁻ ions adsorption in the bilayer lipid surface are presented. In Model I and Model II, the surface is continuous with uniformly distributed functional groups constituting the centres of H⁺ and OH⁻ ions adsorption while in the other the surface is built of lipid molecules, free or with attached H⁺ and OH⁻ ions. In these models contribution of the individual lipid molecule forms to interfacial tension of the bilayer were assumed to be additive. In Model III the adsorption of the H⁺ and OH⁻ ions at the PC–PS bilayer surface was described in terms of the Gibbs isotherm. Theoretical equations are derived to describe this dependence in the whole pH range.     

Sterilization of soybean cotyledons by boiling in lactic acid solution

F. RUÍZ-TERÁN AND J.D. OWENS. 1996. The effect of pH on the heat resistance of Bacillus stearothermophilus spores at 100°C in the presence of 0.11 mol 1^-1 lactic acid and 0.2 mol 1^-1 sodium phosphate buffer was examined. At pH values of 7.0 and 6.0 spores survived 60 min exposure unharmed but at pH 4.3 and 3.0 they died with decimal reduction times (DRTs) of 27 min and 2.8 min, respectively. Death rates were similar in the presence or absence of hydrated soybean cotyledons. In the presence of phosphate buffer and cotyledons at mean pH 3.6 the DRT was 118 min but in the presence, in addition, of lactic acid it was 11 min. It is suggested that the enhanced death rate was due to toxic effects of undissociated lactic acid. Rhizopus oligosporus NRRL 2710 grew well on cotyledons, having pH values from 7.0 to 3.7, prepared by boiling for 60 min in the presence of 0.11 mol 1^-1 lactic acid and 0.2 mol 1^-1 phosphate buffer.     

Research note : Unsuitability of the MRS medium for the screening of hydrogen peroxide-producing lactic acid bacteria

The effect of MRS broth on the stability of hydrogen peroxide (H₂O₂) has been studied. Known concentrations (1–100 μg ml⁻¹) of H₂O₂ were prepared in distilled water, phosphate buffer (pH 7·0) and MRS broth (pH 6·2 and 3·9). H₂O₂ was very stable in aqueous and buffer solutions but it was rapidly degraded in MRS broth (pH 3·9). The presence of H₂O₂ in MRS broth (pH 6·2) could not be detected.     

Functionalization of pine needles by carboxymethylation and network formation for use as supports in the adsorption of Cr

Pine needles and their carboxymethyl forms were functionalized by network formation with 2-acrylamido-2-methylpropanesulphonic acid (AAmPSA) in the presence of N,N-methylene bisacrylamide. N-Tetramethylethylene diamine and ammonium persulfate were used as accelerator-initiator systems to prepare these hydrogels. The hydrogels were characterized by FTIR, SEM, and nitrogen analysis and for water uptake capacities before and after metal ion sorption with a view to evaluating their use in the removal of toxic ionic species from waste water. A detailed study of Cr⁶⁺ adsorption was carried out as a function of time, temperature, pH, and ionic strength. The thermodynamic parameters of adsorption such as ΔH⁰, ΔS⁰, and ΔG⁰ have been evaluated to understand the underlying mechanism of adsorption. In order to understand their reusability in possible technological applications, biodegradability of these hydrogels and their precursors was studied.     

Spectacular abundance of ciliates in anoxic pond water: contribution of symbiont photosynthesis to host respiratory oxygen requirements

Abstract: Very large numbers (3466 ml⁻¹) of ciliated protozoa were found living beneath the oxic-anoxic boundary in a stratified freshwater pond. Most ciliates (96%) contained symbiotic algae ( Chlorella spp.). Peak abundance was in anoxic water with almost 1 mol free CO₂ m⁻³ and a midday irradiance of 6 μmol photon m⁻² s⁻¹. Photosynthetic rate measurements of metalimnetic water indicated a light compensation point of 1.7 μmol photon m⁻² s⁻¹ which represents 0.6% of sub-surface light. We calculate that photosynthetic evolution of O₂ by symbionts is sufficient to meet the demand of the host ciliates for 13 to 14 hours each day. Each 'photosynthetic ciliate' may therefore become an aerobic island surrounded by anoxic water.     

牡丹根际溶磷放线菌的筛选及其溶磷特性

通过从牡丹根际土壤中分离筛选溶磷放线菌,得到一株具有较强溶磷能力的菌株PSPSA1,根据形态特征、生理生化特性以及16S rDNA序列分析对菌株进行鉴定,并研究其溶磷遗传稳定性及溶磷特性.菌株PSPSA1被鉴定为白网链霉菌,具有较好的溶磷遗传稳定性.在不同磷源培养液中溶磷量依次为磷酸钙(158.5 mg·L^-1)>磷酸铝(139.9 mg·L^-1)>磷酸铁(127.7 mg·L^-1)>卵磷脂(45.6 mg·L^-1),在无机磷培养液中的溶磷量均与pH呈现显著负相关性,在有机磷培养液中的溶磷量与pH没有显著相关性.在不同碳源条件下的溶磷量依次为乳糖>葡糖糖>麦芽糖>果糖>蔗糖>淀粉>纤维素,在不同氮源条件下的溶磷量依次为蛋白胨>硝酸铵>硫酸铵>硝酸钾>尿素,以葡萄糖为碳源、蛋白胨为氮源时,菌株的溶磷量最高可达202.6 mg·L^-1.土培60 d,单施菌株土壤有效磷含量比对照增加68.2%,菌株与有机肥混施土壤有效磷含量比单施有机肥增加76.7%.表明菌株PSPSA1能够溶解多种难溶磷,在土壤中溶磷效果显著,与有机肥混施其溶磷能力明显提高,有望成为高效生物磷肥的优良菌种.