Properties of lead deposits in cell walls of radish (Raphanus sativus) roots

Various mechanisms are involved in detoxification of heavy metals such as lead (Pb) in plant cells. Most of the Pb taken up by plants accumulates in their roots. However, the detailed properties of Pb complexes in roots remain unclear. We have investigated the properties of Pb deposits in root cell walls of radish (Raphanus sativus L.) seedlings grown on glass beads bed containing Pb pellets, which are the source of Pb-contamination in shooting range soils. Pb deposits were tightly bound to cell walls. Cell wall fragments containing about 50,000 ppm Pb were prepared from the roots. After extracting Pb from the cell wall fragments using HCl, Pb ions were recombined with the Pb-extracted cell wall fragments in a solution containing Pb acetate. When the cell wall fragments were treated with pectinase (E.C. 3.2.1.15) and were chemically modified with 1-ethyl-3-dimethylamino-propylcarboimide, the Pb-rebinding ability of the treated cell wall fragments decreased. When acid-treated cell wall fragments were incubated in a solution containing Pb²⁺ and excess amounts of a chelating agent, Pb recombined with the cell wall fragments were measured to estimate the affinity between Pb²⁺ and the cell wall fragments. Our data show that Pb²⁺ binds to carboxyl groups of cell walls. The source of the carboxyl groups is suggested to be pectic compounds. A stability constant of the Pb-cell wall complex was estimated to be about 10⁸. The role of root cell walls in the mechanism underlying heavy metal tolerance was discussed.     

Microalgal-luffa sponge immobilized disc: a new efficient biosorbent for the removal of Ni(II) from aqueous solution

AIMS: The aim was to develop a new, efficient and cost-effective biosorbent for the removal of heavy metals from aqueous solution. METHODS AND RESULTS: A new biosorbent was developed by immobilizing a unicellular green microalga Chlorella sorokiniana within luffa sponge discs and used for the removal of metal ions from aqueous solution. Microalgal-luffa sponge immobilized discs (MLIDs) removed Ni(II) very rapidly, with 97% of equilibrium loading being reached in 5 min. MLIDs were tested for their potential to remove Ni(II) from aqueous solution in fixed-bed column bioreactor. The regenerated MLIDs retained 92.9% of the initial binding capacity for Ni(II) up to five cycles of reuse. CONCLUSIONS: In this study for the first time, C. sorokiniana biomass immobilized within luffa sponge disc was successfully used as a metal biosorbent for the removal of Ni(II). It appears that MLIDs can be used as an effective biosorbent for efficient removal of Ni(II) or other metals from aqueous solution. SIGNIFICANCE AND IMPACT OF THE STUDY: MLIDs biosorption system was shown to have good biosorption properties with respect to Ni(II). Efficient metal removal ability of MLIDs, low cost and simplicity of the technique used for the preparation of MILDs could provide an attractive strategy for developing high-affinity biosorption system for heavy metal removal.     

Copper sorption by native and modified pellets of wood-rotting basidiomycetes

AIMS: The aim of this study was to investigate the biosorption of copper to the pellets of different wood-rotting fungal species. METHODS AND RESULTS: Copper sorption was studied in both batch and column arrangements. The optimum pH for copper sorption was between 3.5 and 4. In 100 mg l(-1) Cu (II), maximum qe values were found for Oudemansiella mucida (8.77 mg g(-1) dry wt), Lepista nuda (6.29 mg g(-1)), Pycnoporus cinnabarinus (5.08 mg g(-1)) and Pleurotus ostreatus (4.77 mg g(-1)). Both biomass yield and specific sorption were influenced by the composition of the fermentation broth. The results of column experiments showed that mycelial pellets of wood-rotting fungi can be considered as promising biosorbent material. CONCLUSIONS: Pellets of wood-rotting fungi showed the same or better copper sorption properties as those previously reported for lower fungi or filamentous bacteria, as well as good mechanical properties.     

Heavy chain of human high molecular weight and low molecular weight kininogens binds calcium ion

An antibody subpopulation, anti high molecular weight (anti-HMW) kininogen-Ca2+ antibody able to bind specifically to the HMW kininogen-Ca2+ complex, was isolated from anti-HMW kininogen antiserum. Partially purified anti-HMW kininogen antibody was applied to a HMW kininogen-Sepharose column equilibrated with 40 mM tris(hydroxymethyl)aminomethane hydrochloride buffer, pH 7.5, containing 1.0 M NaCl and 1 mM CaCl2, and anti-HMW kininogen-Ca2+ antibody was eluted with 5 mM ethylenediaminetetraacetic acid. As a result of characterization by enzyme-linked immunosorbent assay, this antibody specifically recognized the cyanogen bromide cleaved fragment 1 (CB-1) region (1-160 amino acid sequence) of the heavy chain of kininogen molecules in the presence of Ca2+ or Mg2+. Furthermore, circular dichroism (CD) experiments showed that the conformational changes of HMW kininogen and heavy chain were induced by metal ions such as Ca2+ and Mg2+ and that these changes were due to the conformational change of the CB-1 region of the heavy chain. The dissociation constant (Kd) for the heavy chain-Ca2+ measured by CD analysis at 214 nm was found to be 0.33 +/- 0.09 mM (mean +/- SD). The number of Ca2+-binding sites of heavy chain calculated from the Hill plot was 1.15 +/- 0.04 (mean +/- SD). Then, a possible Ca2+-binding site was found in the amino-terminal portion of the heavy chain of kininogen molecules.     

High-performance analytical applications of immobilized metal ion affinity chromatography

The separation of three sets of standard protein mixtures on a high-performance immobilized metal ion affinity chromatography (HP-IMAC) column by elution with linear gradients of imidazole is described. The affinity of the test proteins for the immobilized metal ions follows the order Cu2+ greater than Ni2+ greater than Zn2+. The iminodiacetic acid-Cu2+ column gives the best resolution of all three protein mixtures and is the only immobilized metal ion column that can be used for elution of absorbed proteins with a decreasing pH gradient. An application of HP-IMAC for the separation of monoclonal IgG from mouse ascites fluid is also outlined. This versatile separation method is thus suitable for both analytical and preparative separations of proteins and peptides resulting in high recoveries and good reproducibility. The leakage of immobilized metal ions from the TSK gel chelate-5PW is apparent if the column is eluted by buffers containing low concentrations of (i) glycine or (ii) primary amines at round neutral pH. Considerable amounts of immobilized Zn2+ and Ni2+ ions also leak from the column by washing with buffers of pH 4.5 or lower. However, all three immobilized metal ions are stable toward exposure to low concentrations of imidazole (up to 50 mM) in phosphate buffers between pH 6.5 and 8.0. Adsorbed proteins could thus be eluted conveniently by using linear gradients of imidazole to give reproducible results. Moreover, this elution procedure made it possible to use the IMAC columns for repeated runs without the need for regeneration and recharging of the columns with fresh metal ions after each use.     

Removal of heavy metals from aqueous solution using Rhizopus delemar mycelia in free and polyurethane-bound form

This study assesses the ability of mycelia of Rhizopus delemar (both free and immobilized on polyurethane foam) to remove heavy metals from single-ion solutions as well as from a mixture of them. All experiments were conducted using 0.5-5 mm solutions of CuSO4 x 5H2O, CoCl2-6H2O and FeSO4 7H2O. Mycelia immobilized on polyurethane foam cells showed some times increase in uptake compared with that of free cells. Metal ions accumulation from a mixed solution was decreased slightly for cobalt and iron and considerable for copper ions. Heavy metal uptake was examined in the immobilized column experiments and more than 92% heavy metal removal (mg heavy metals removed/mg heavy metals added) from a mixed solution was achieved during the 5 cycles. During these experiments, the dry weight of the immobilized cells was decreased by only 2%. These results showed that immobilized mycelia of Rhizopus delemar can be used repeatedly for removal of heavy metals from aqueous solutions.     

沉水植物菹草在低温条件下对重金属Cu、Pb、Zn的吸附和富集

在10℃的较低温度条件下,研究了冬春季节生长旺盛的沉水植物菹草(Potamogeton crispus L.)对重金属离子Cu²⁺,Pb²⁺,Zn²⁺的生物吸附特征及解吸情况,对不同初始浓度重金属水体中的重金属离子去除率情况,以及在此过程中菹草各器官(叶、茎、根茎、根)对重金属离子的富集情况。结果表明,菹草对Cu²⁺,Zn²⁺的吸附在20 min内达到平衡,对Pb²⁺的吸附在50 min内达到平衡,吸附动力学结果符合伪二级动力学方程,决定系数分别达1,1,0.997 8。Freundlich等温线可较好地拟合菹草吸附Cu²⁺,Pb²⁺,Zn²⁺的过程,Cu²⁺,Pb²⁺,Zn²⁺的吸附容量分别达到66.900 6,26.543 0,30.371 8 mg·L^-1。以去离子水作洗脱剂,解吸液中3种重金属离子浓度均低于仪器检出限(0.01 mg·L^-1),解吸程度微弱。投放菹草后,随着初始处理浓度的升高,水体Cu²⁺的去除率先降低后升高,Pb²⁺的去除率的变化趋势与Cu²⁺类似。Zn²⁺去除率则随水体Zn²⁺初始浓度的升高而逐渐升高。菹草各器官对水体3种重金属离子的富集能力不同,排序为Cu²⁺＞Zn²⁺＞Pb²⁺。不同器官对同一种重金属离子的富集量差异显著,叶是富集重金属离子的主要器官。水体重金属离子的初始浓度会影响菹草各器官富集重金属离子的能力,一般随水体重金属初始浓度升高,菹草各器官的重金属离子富集量虽有不同程度的增加但富集系数持续减小。     

Biochemical oxygen demand sensor using Serratia marcescens LSY 4

A microbial biochemical oxygen demand (BOD) sensor consisting of Serratia marcescens LSY 4 and an oxygen electrode was prepared for estimation of the biochemical oxygen demand. The response of the BOD sensor was insensitive to pH in the range of pH 6.0-8.0, and the baseline drift of the signal was nearly absent even in unbuffered aqueous solution. Because heavy metal ions were precipitated from the phosphate buffer solution, unbuffered solution was used to investigate the effect of the concentration of heavy metal ions on the sensor response. Contrary to previous studies, not only Cu2+ and Ag+ but also Cd2+ and Zn2+ significantly decreased the response of the BOD sensor in unbuffered solution. Graft polymerization of sodium styrene sulfonate on the surface of the porous teflon membrane was carried out to absorb the heavy metal ions permeating through the membrane. Tolerance against Zn2+ was induced for S. marcescens LSY 4 to make the cells less sensitive to the presence of heavy metal ions. The membrane modification and the Zn2+ tolerance induction showed some positive effects in such a way that they reduced the inhibitory effects of Zn2+ and Cd2+ on the sensitivity of the BOD sensor. However, they had no effect on the protection of the cells against the interference of Cu2+ and Ag+ on the performance of the sensor.     

一株红假单胞菌的分离及对Cu2+的去除

【背景】重金属污染对环境和人类的健康构成了重大威胁,因此,重金属污染的治理迫在眉睫。生物治理因成本低、处理效果好和无二次污染等优点,在处理重金属污染时被优先选择。【目的】从辽河入海口深13 m的水体中,利用紫色非硫细菌富集培养基筛选产色素能力强并对高浓度Cu~(2+)有高效去除能力的菌株。【方法】采用形态学、生理生化特性和分子生物学方法鉴定菌种;采用二乙基二硫代氨基甲酸钠分光光度法测定Cu~(2+)的含量。【结果】鉴定菌株为红假单胞菌属,将其命名为Rhodopseudomonas sp. gh32。菌株gh32的最适生长温度和最适pH分别为30°C和7.0,其在pH 5.0-10.0范围内可正常生长,在3 mmol/L的CuSO4溶液中能够正常生长,能利用葡萄糖、甘露糖和果糖等单糖和硫化氢。菌株gh32在24h内对Cu~(2+)的去除率均在99%以上,处理能力为1 331 g/g干菌重或167.6 g/g湿菌重。【结论】菌株gh32对Cu~(2+)具有较强的耐受性和很好的去除效果,是治理含Cu~(2+)废水的潜力菌株。本研究为生物治理重金属废水提供了支持。     

Influence of heavy metal ions on the electrophysical properties of Anacystis nidulans and Escherichia coli cells]

The influence of heavy metal ions (Ag+, Cu2+, Cd2+, Pb2+, Mn2+, Zn2+, Gd3+, 1 microM-1 mM) on Anacystis nidulans and Escherichia coli cells has been studied by means of electrophoresis and electro-orientation spectroscopy methods. It has been shown that changes of cell electrophoretic mobility (EM) and low-frequency (20 Hz) electro-orientation effect (EOE) observed with the increase of metal cation concentration characterize the adsorption of these ions on surface layers of cell envelopes. The degree and the character of these changes depend on cation valency and the initial value of cell EM. At the same time different changes of EM and EOE as a result of the multivalent cation adsorption allows to conclude that in that case the anisotropy of the cell surface increases. Cell damages were determined by changes in high-frequency EOE of cells which indicated the disturbance of barrier properties of their cytoplasmic membrane. Toxic effects of Ag+, Cu2+, Cd2+ ions on cells of both species and of Pb2+ on E.coli cells were observed. By toxic effects on the cytoplasmic membrane these ions could be placed in the order: for A.nidulans cells--Ag+ greater than Cu2+ greater than Cd2+; for E.coli cells Ag+ greater than Cu2+ greater than Cd2+ greater than Pb2+. Higher toxicity of heavy metals on E.coli cells seems to be connected with the more negative charge of deep layers of the cell surface.     

重金属 Ni2+ 对亚洲玉米螟生长发育和生殖行为的影响

【目的】探索农田重金属Ni²⁺污染对植食性昆虫潜在的影响。【方法】在人工饲料中添加不同浓度（1, 5, 10, 15和20 mg/kg）的重金属Ni²⁺,以不添加重金属的人工饲料为对照,实验室条件下饲养亚洲玉米螟 Ostrinia furnacalis (Guenée)初孵幼虫,系统观察其各项生长发育和生殖行为指标参数。【结果】Ni ²⁺对亚洲玉米螟幼虫期、蛹期、化蛹率和羽化率有抑制作用;对亚洲玉米螟雌蛹重有促进作用,但随Ni²⁺浓度的升高而逐渐减弱;低浓度Ni²⁺（1~10 mg/kg）对雄蛹重有促进作用,高浓度Ni²⁺（15~20 mg/kg）对雄蛹重有抑制作用。随着Ni²⁺浓度的增加,亚洲玉米螟雌蛾平均求偶持续时间逐渐减小;1 mg/kg Ni²⁺浓度下雌蛾求偶率及求偶高峰期出现的时间与对照组基本一致,其余4个Ni²⁺浓度都对雌蛾求偶行为产生影响且表现为求偶高峰期出现的时间提前,最高求偶率均小于100%。5 mg/kg Ni²⁺对亚洲玉米螟雄蛾定向行为有促进作用,其余浓度则表现为抑制作用。Ni²⁺对亚洲玉米螟雌蛾的产卵量和卵孵化率均产生了显著的抑制作用。【结论】重金属Ni²⁺对亚洲玉米螟的生长发育和生殖行为产生了影响。     

Accumulation and removal of heavy metal ions by insolubilized-DNA and its interaction.

DNA was immobilized onto a porous glass bead by a treatment with UV irradiation. The immobilized DNA was insoluble in water and used for accumulation of heavy metal ion. When DNA-immobilized glass bead was added into aqueous solution containing heavy metal ions, such as Hg2+, Cd2+, Pb2+, Zn2+, Cu2+ and Fe3+, the concentration of these metal ions in the solution was decreased. However, the concentration of Mg2+ in the solution was not affected by the addition of the DNA-immobilized glass bead. These results suggested that UV-irradiated DNA selectively accumulated heavy metal ions.     

新月弯孢霉菌丝球对染料脱色作用的研究

研究了新月弯孢霉Curvularia lunata JQH-100液体培养时产生的菌丝球对多种染料的脱色能力。结果表明,多种染料在24h内的脱色率均达到80%以上,且菌丝球稳定性良好,可重复使用6次;以菌丝球对孔雀绿脱色效果为优化指标,正交实验优化获得制备菌丝球的最佳条件为:葡萄糖20g/L、硫酸铵5g/L、马铃薯200g/L、KH2PO43g/L、MgSO45mg/L、CuSO40.5mg/L、VB15mg/L及pH5、摇床转速120r/min。在上述优化后的基础培养基(不含MgSO4、CuSO4)中分别添加微量元素Cu2+、Mn2+、Mg2+或Ca2+制备的菌丝球,对孔雀绿脱色能力增强;添加Fe2+制备的菌丝球,对孔雀绿脱色能力下降;分别添加Zn2+、Al3+或Na+制备的菌丝球对孔雀绿脱色能力与对照相近。应用优化后培养条件制备的菌丝球处理含多种染料的混合废水,也获得了较好的脱色效果。     

Isolation and Molecular Characterization of Heavy Metal-Resistant Alcaligenes faecalis from Sewage Wastewater and Synthesis of Silver Nanoparticles

Environmental pollution with toxic heavy metals is increasing throughout the world alongside industrial development. Microorganisms and microbial products can be highly efficient bioaccumulators of soluble and particulate forms of metals, especially dilute external solutions. Microbe related technologies (Biotechnology) may provide an alternative or additive conventional method for metal removal or metal recovery. This study dealt with isolation, identification and characterization of heavy metal-resistant (Pb²⁺, Cd²⁺, Al³⁺, Cu²⁺, Ag²⁺ and Sn²⁺) bacteria from sewage wastewater at Taif Province, Saudi Arabia. Nine bacterial isolates were selected by using an enrichment isolation procedure based on high level of heavy metal resistance. All the isolates showed high resistance to heavy metals with Minimum Inhibitor Concentration (MIC) ranging from 800 μg/ml to 1400 μg/ml. All nine resistant isolates showed multiple tolerances to heavy metals. On the basis of morphological, biochemical and 16S rRNA characterization, the most potent isolates (Cd1-1, Ag1-1, Ag1-3 and Sn1-1) were identified as Alcaligenes faecalis. Scanning electron microscope analysis showed that the morphology of Alcaligenes faecalis Ag1-1 was unchanged after growth in medium without and with addition of Ag²⁺ indicative Ag²⁺ is not toxic to the isolate under the conditions tested. The ability of Alcaligenes faecalis Ag1-1 to synthesize sliver nanoparticles was examined. The heavy metal-resistant bacteria obtained could be useful for the bioremediation of heavy metal-contaminated environment.     

Physical Studies on Cell Immobilization Using Calcium Alginate Gels

Columns of calcium alginate gel pellets have excellent physical properties when used as a cell immobilization support. Columns of pellets were very resistant to compression and abrasion during passage of high concentrations of sucrose at high flow rates, but if the pellets were formed using low alginate and Ca²⁺ concentrations, compression occurred and flow out of the column was reduced and pressure built up. Transfer of sucrose into the pellets was controlled by internal diffusion, the rate of diffusion being increased by reductions in the alginate and Ca²⁺ concentrations used for immobilization and by the presence of entrapped active cells. Some leakage of cells occurred during use especially when cell division of the entrapped cells took place, but leakage could be minimized by using more highly polymerized pellets. Therefore, immobilization conditions can be chosen so as to form strong pellets, possessing high substrate transfer rates and low rates of cell leakage.     

小麦不同生育时期Cd、Cr、Pb污染监测指标体系

以小麦为供试材料,分别采用发芽试验、溶液培养、土柱栽培等毒理试验方法,研究了小麦萌芽期、幼苗期及成株期受重金属污染毒害的指标体系。结果表明,①小麦根伸长抑制率可作为萌芽期重金属污染评价的一项生物指标;Cd、Cr、Pb对小麦根生长的7d半效应浓度(EC50)值分别为1.39、0.20mmo.lL-1和2.75mmol.L-1,据此得到3种重金属对小麦同一性状的毒性次序为CrCdPb;此外各性状抑制率与胁迫浓度的关系可用双曲线模型y=x/(a+x/100)或指数曲线模型y=a(1-exp(-bx))较好模拟。②Cd、Cr、Pb单一污染胁迫对小麦幼苗性状的毒性次序均以影响叶面积和冠部干重为主,表明叶面积和冠部干重为幼苗期污染监测的敏感指标;重金属对除根干重外所有性状的毒性次序:CrCdPb,该毒性次序是根据引起50%抑制的临界浓度即EC50值确定的;各性状抑制率与胁迫浓度的关系可用直线模型、或双曲线模型、或指数曲线模型较好模拟。③重金属单一污染胁迫造成小麦籽粒产量下降幅度是CdCrPb;二元互作对产量影响的重要性次序为CdCrCdPbCrPb;在Cd、Cr、Pb复合污染胁迫条件下,穗数可以作为成株期重金属污染监测的首选指标。以上指标可为农田小麦重金属污染的监测和综合治理提供一些理论依据。     

Heavy metal resistant freshwater ciliate, Euplotes mutabilis, isolated from industrial effluents has potential to decontaminate wastewater of toxic metals

The ciliate, Euplotes mutabilis, isolated from industrial wastewater of tanneries of Kasur, Pakistan, showed tolerance against Cd2+ (22 microg ml(-1)), Cr6+ (60 microg ml(-1)), Pb2+ (75 microg ml(-1)) and Cu2+ (22 microg ml(-1)). The heavy metals, Cr and Pb, were randomly selected for determining the capability of the ciliate to reduce the concentration of these metal ions in the medium and to evaluate its potential use as bioremediator of wastewater. The live protozoans could remove 97% of Pb2+ and 98% of Cr6+ from the medium, 96 h after inoculation of the medium containing 10 micro gml(-1) of metal ions. The acid digestion of ciliate showed 89% of Pb2+ and 93% of Cr6+ ions accumulated in the organism. When the ciliate was exposed to heavy metals at a larger scale viz., 10 l of water containing 10 micro gml(-1) of heavy metals, it removed 86% of Pb2+ and 90% of Cr6+ from the medium. The metal uptake ability of E. mutabilis, as evidenced by its survival and growth in 100ml and 10 l of water containing 10 microg ml(-1) of metal ions, reduction in the concentration of heavy metals in the medium and its increased uptake by the live cells, and no metal uptake by the heat killed ciliate can be exploited for metal detoxification of industrial wastes and environmental clean-up operations.     

Effect of heavy metals on the glutathione status in different ectomycorrhizal <Emphasis Type="Italic">Paxillus involutus</Emphasis> strains

The glutathione (GSH) status and heavy metal tolerance were investigated in four Paxillus involutus strains isolated from different heavy-metal-polluted and non-polluted regions of Europe. The heavy metal burden in the habitats did not affect significantly either the heavy metal (Cr₂O₇²⁻, Cd²⁺, Hg²⁺, Pb²⁺, Zn²⁺, Cu²⁺) tolerance and accumulation or the GSH production of the strains tested. Exposures to heavy metals increased the intracellular GSH concentrations in 12 from 24 experimental arrangements (four strains exposed to six heavy metals) independently of the habitats of the strains. The importance of GSH in heavy metal tolerance (high MIC values, ability to accumulate heavy metals and to grow in the presence of heavy metals) was thus demonstrated in this ectomycorrhizal fungus.     

Cadmium biosorption properties of the metal‐resistant Ochrobactrum cytisi Azn6.2

The aim of this work was to establish the conditions for using Ochrobactrum cytisi Azn6.2 as a metal biosorbent. Azn6.2 is a novel strain from the legume symbiont O. cytisi that has been isolated from nodules of Medicago polymorpha plants grown on heavy metal‐polluted soils. Compared with the strain ESC1, Azn6.2 showed some biochemical differences, as well as antibiotic susceptibility, Azn6.2 was multi‐resistant to heavy metals, such as Cu, Cd and Zn, and bacterial pellets were able to biosorb high amounts of Cd and Zn. As shown by scanning electron microscopy coupled to energy dispersive X‐ray, most of Cd was attached to the cell surface. Optimal conditions for Cd biosorption were established, being 1 mM Cd ions in solution and 2 h of contact with the biosorbent at room temperature. At these conditions, maximal Cd loading capacity reached 32–34 mg/g. Cd desorption from bacterial pellets was achieved after washing with EDTA or, at higher efficiency, at pH 1.0. These results indicated that biosorption/desorption on O. cytisi Azn6.2 biomass should be a cost‐effective method for Cd recovery from contaminated solutions.     

Purification of a 51 kDa endo-β-N-acetylglucosaminidase from Staphylococcus aureus

A bacteriolytic enzyme obtained from the culture fluid of Staphylococcus aureus FDA 209P was purified to homogeneity utilizing dye-ligand affinity column chromatography, hydrophobic interaction high pressure liquid chromatography (HPLC) and hydroxyapatite HPLC. Subsequent characterizations indicated that the purified enzyme acted as endo-beta-N-acetylglucosaminidase. The molecular weight determined by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) was 51,000 and the isoelectric point was higher than 10. The optimum pH for the enzyme activity on whole cells of Micrococcus luteus as a substrate was 8.0. Some heavy metal cations (Cu2+ and Zn2+) inhibited the enzyme activity at a concentration of 0.1 mM and others (Ba2+, Mg2+ and Co2+) showed a stimulating effect at a concentration of 1 mM.