Functional comparision between truncated MTT1 and truncated MTT2 from Tetrahyemna thermophila

Metallothioneins (MTs) are low-molecular-weight proteins with high Cys content and high metal-chelating ability. CdMT and CuMT subfamilies present different characteristics in Tetrahymena. To explore the effect of the cysteine arrangement and sequence length of MTs for binding different metal ions, MTT1, truncated MTT1 (TM1), MTT2, and truncated MTT2 (TM2) were expressed in E. coli. The half-maximal inhibiting concentrations (IC₅₀) of Cd²⁺ and Cu⁺ for the recombinant strains were different. Furthermore, E. coli cells expressing MTT1 and TM1 exhibited higher accumulating ability for Cd²⁺ than cells expressing MTT2 and TM2. However, the opposite is true for Cu⁺. The binding ability of the different recombinant proteins to Cd²⁺ and Cu⁺ were also different. MTT1 and truncated mutant TM1 were the preference for Cd²⁺, whereas MTT2 and truncated mutant TM2 were the preference for Cu⁺ coordination. These results showed that metal ion tolerance and accumulation ability not only depended on cysteine arrangement pattern but also on sequence length of MT in Tetrahymena.     

Hints for Metal-Preference Protein Sequence Determinants: Different Metal Binding Features of the Five Tetrahymena thermophila Metallothioneins

The metal binding preference of metallothioneins (MTs) groups them in two extreme subsets, the Zn/Cd- and the Cu-thioneins. Ciliates harbor the largest MT gene/protein family reported so far, including 5 paralogs that exhibit relatively low sequence similarity, excepting MTT2 and MTT4. In Tetrahymena thermophila, three MTs (MTT1, MTT3 and MTT5) were considered Cd-thioneins and two (MTT2 and MTT4) Cu-thioneins, according to gene expression inducibility and phylogenetic analysis. In this study, the metal-binding abilities of the five MTT proteins were characterized, to obtain information about the folding and stability of their cognate- and non-cognate metal complexes, and to characterize the T. thermophila MT system at protein level. Hence, the five MTTs were recombinantly synthesized as Zn²⁺-, Cd²⁺- or Cu⁺-complexes, which were analyzed by electrospray mass spectrometry (ESI-MS), circular dichroism (CD), and UV-vis spectrophotometry. Among the Cd-thioneins, MTT1 and MTT5 were optimal for Cd²⁺ coordination, yielding unique Cd₁₇- and Cd₈- complexes, respectively. When binding Zn²⁺, they rendered a mixture of Zn-species. Only MTT5 was capable to coordinate Cu⁺, although yielding heteronuclear Zn-, Cu-species or highly unstable Cu-homometallic species. MTT3 exhibited poor binding abilities both for Cd²⁺ and for Cu⁺, and although not optimally, it yielded the best result when coordinating Zn²⁺. The two Cu-thioneins, MTT2 and MTT4 isoforms formed homometallic Cu-complexes (major Cu₂₀-MTT) upon synthesis in Cu-supplemented hosts. Contrarily, they were unable to fold into stable Cd-complexes, while Zn-MTT species were only recovered for MTT4 (major Zn₁₀-MTT4). Thus, the metal binding preferences of the five T. thermophila MTs correlate well with their previous classification as Cd- and Cu-thioneins, and globally, they can be classified from Zn/Cd- to Cu-thioneins according to the gradation: MTT1>MTT5>MTT3>MTT4>MTT2. The main mechanisms underlying the evolution and specialization of the MTT metal binding preferences may have been internal tandem duplications, presence of doublet and triplet Cys patterns in Zn/Cd-thioneins, and optimization of site specific amino acid determinants (Lys for Zn/Cd- and Asn for Cu-coordination).     

Saccharomyces cerevisiae and Neurospora crassa contain heavy metal sequestering phytochelatin

In fungi, cellular resistance to heavy metal cytotoxicity is mediated either by binding of metal ions to proteins of the metallothionein type or by chelation to phytochelatin-peptides of the general formula (-Glu-Cys)_n-Gly. Hitherto, only one fungus, Candida glabrata has been shown to contain both metal inactivating systems. Here we show by unambiguous FAB-MS analysis that both a metallothionein-free mutant of Saccharomyces cerevisiae as well as a wildtype strain synthesize phytochelatin (PC₂) upon exposure to 250 M Cd²⁺ ions. The presence of Zn and/or Cu ions in the nutrient broth also induces PC₂ synthesis in this organism. By ¹⁰⁹Cd exchange and subsequent monobromobimane fluorescence HPLC, it could be shown that the presence of Cd²⁺ in the growth medium also induces phytochelatin synthesis in Neurospora crassa, which contains metallothioneins.     

Differences in the binding modes of phytochelatin to cadmium (II) and Zinc(II) ions

An ¹H NMR (nuclear magnetic resonance) spectroscopic structural analysis of Cd²⁺ complexes formed with the pentapeptide phytochelatin, (NH₃)⁺−(ψ-Glu-Cys)₂−Gly−COO−(PC₂), at a pH of 7.5 showed that the two thiol groups of the Cys residues and either the carbonyl or amide group of the peptide bond between Glu1 and Cys1 act as possible donor groups in the complexes at Cd²⁺/PC₂ ratios up to 0.4. As the ratio increases, the carboxylate group of Glu2 and either the carbonyl or amide group of the peptide bond between Cys1 and Glu2 comes to serve as a donor group. The manner in which Cd²⁺ forms complexes with PC₂ is distinctly different from Zn²⁺ and might account for the role of phytochelatin in Cd²⁺ detoxification. Electron absorption spectrometry demonstrated that the Cd²⁺ complexes are coordinated in a tetrahedral fashion by four thiol groups and that several sulfur atoms might bridge Cd²⁺ ions, resulting in the formation of polynuclear complexes. This contrasts with Zn²⁺ complex formation, which consists exclusively of a 1:1 complex.     

Metal resistance and removal by two strains of the green alga, <Emphasis Type="Italic">Chlorella vulgaris</Emphasis> Beijerinck,isolated from Laguna de Bay,Philippines

Two strains of Chlorella vulgaris Beijerinck isolated from two different sites in Laguna de Bay, Philippines, were studied for their resistance and ability to remove four metal ions, i.e., Cu²⁺, Cr⁶⁺, Pb²⁺, and Cd²⁺ added separately in BG-11 growth medium. The growth of the two strains was severely inhibited at 2 mg.L⁻¹ of Cu²⁺, 5 mg.L⁻¹ of Cr⁶⁺, 8 mg.L⁻¹ of Pb²⁺, and 10 mg.L⁻¹ of Cd²⁺. However, the two strains exhibited different EC₅₀ values for the same metal ion. The WB strain had a significantly higher resistance (p < 0.01) for Cd²⁺ and Cr⁶⁺ compared with the SB strain, while the SB strain had significantly higher resistance (p < 0.01) for Cu²⁺ compared with the WB strain. On the other hand, the two strains behaved differently in their capacity to remove the metal ions in BG-11 medium containing 1.0 mg.L⁻¹ of the three metal ions, except for Cu²⁺, which was added at 0.1 mg.L⁻¹. The WB strain showed the highest removal of Cd²⁺ at 70.3% of total, followed by Pb²⁺ at 32%, while the SB strain exhibited the highest removal of Pb²⁺ at 48.7% followed by Cd²⁺ at 40.7% of the total. Both strains showed the least removal of Cr⁶⁺ at 28% and 20.8% of the total for the WB and SB strains respectively. The percentage removal for Cu²⁺ was 50.7% and 60.8% for the WB and SB strains respectively. After 12 days of incubation, both strains showed that a greater percentage of the metal ions removed were accumulated intracellularly than adsorbed at a ratio of at least 2:1. Both strains manifested the same cytological deformities, like a loss of pyrenoids at 10 mg.L⁻¹ in all four metal ions. Discoloration and disintegration of chloroplasts were observed at 1.0 mg.L⁻¹ in Cu²⁺ and 5 mg.L⁻¹ in Cr⁶⁺. The nonrelease of autospores from the mother cells was observed at 10 mg.L⁻¹ in Cu²⁺ and Cr⁶⁺. Presented at the 6th Meeting of the Asian Pacific Society of Applied Phycology, Manila, Philippines.     

Differential action of Hg2+ and Cd2+ on the phycobilisomes and chlorophyll a fluorescence,and photosystem II dependent electron transport in the cyanobacterium Anabaena flos-aquae

The effect of equimolar concentrations of Hg²⁺ and Cd²⁺ on the whole cell absorption spectra, absorption spectra of the extracted phycocyanin (PC) and fluorescence emission spectra of phycobilisomes (PBS) was investigated in the cells of Anabaena flos-aquae. The PC component of the PBS was found to be extremely sensitive to the Hg²⁺ rather than the Cd²⁺ ions. Further, the results showed that Hg²⁺ and Cd²⁺ induced decrease in the rate of Hill activity (H₂O - DCPIP) was partially restored by the electron donor NH₂OH, not by the diphenyl carbazide. Similarly, chlorophyll a fluorescence emission in the presence of metals showed that addition of NH₂OH could effectively reverse the metal induced alterations in the fluorescence emission intensity. These results, together, suggested that Hg²⁺ and Cd²⁺ caused damage to the photosystems (PS) II reaction center. However, a relatively higher stimulation of the chlorophyll a emission at 695 nm with a red shift of 4.0 nm in the presence of Hg²⁺, and Cd²⁺ induced preferential decrease in the emission intensity at 676 nm as compared with the peak at 695 nm were indicative of the differential action of Hg²⁺ and Cd²⁺ on the PS II.     

Inhibition of the biosynthesis ofN-acetylneuraminic acid by metal ions and seleniumin vitro

In liver homogenate the biosynthesis ofN-acetylneuraminic acid usingN-acetylglucosamine as precursor can be followed stepwise by applying different chromatographic procedures. In this cell-free system 16 metal ions (Zn²⁺, Mn²⁺, La³⁺, Co²⁺, Cu²⁺, Hg²⁺, VO ₃ ^– , Pb²⁺, Ce³⁺, Cd²⁺, Fe²⁺, Fe³⁺, Al³⁺, Sn²⁺, Cs⁺ and Li⁺) and the selenium compounds, selenium(IV) oxide and sodium selenite, have been checked with respect to their ability to influence a single or possible several steps of the biosynthesis ofN-acetylneuraminic acid. It could be shown that the following enzymes are sensitive to these metal ions (usually applied at a concentration of 1 mmoll^–1):N-acetylglucosamine kinase (inhibited by Zn²⁺ and vandate), UDP-N-acetylglucosamine-2-epimerase (inhibited by zn²⁺, Co²⁺, Cu²⁺, Hg²⁺, VO ₃ ^– , Pb²⁺, Cd²⁺, Fe³⁺, Cs⁺, Li⁺, selenium(IV) oxide and selenite), andN-acetylmannosamine kinase (inhibited by Zn²⁺, Cu²⁺, Cd²⁺, and Co²⁺). Dose dependent measurements have shown that Zn²⁺, Cu²⁺ and selenite are more efficient inhibitors of UDP-N-acetylglucosamine-2-epimerase than vanadate. As for theN-acetylmannosamine kinase inhibition, a decreasing inhibitory effect exists in the following order Zn²⁺, Cd²⁺, Co²⁺ and Cu²⁺. In contrast, La³⁺, Al³⁺ and Mn²⁺ (1 mmoll^–1) did not interfere with the biosynthesis ofN-acetylneuraminic acid. Thus, the conclusion that the inhibitory effect of the metal ions investigated cannot be regarded as simply unspecific is justified.Dedicated to Professor Theodor Günther on the occasion of his 60th birthday     

Ca2+ and Mg2+ ions counteract the reduction by fosetyl-Al (aluminum tris[ethyl phosphonate]) of peroxidase activity from suspension-cultured grapevine cells

Grapevine (Vitis vinifera cv. Monastrell) cell suspension cultures were treated with 1.5 mM fosetyl-Al, a frequently used systemic fungicide for grapevine diseases caused by oomycetes. These cells showed a reduction in the level of peroxidase activity secreted into the culture media when compared to non-treated cells, the effect being mainly related to a decrease in the level of the basic B₁ peroxidase isozyme. The effect of fosetyl-Al on peroxidase was analogous to that observed with the Ca²⁺-channel blockers Co²⁺, Cd²⁺ and La³⁺, and was counteracted by Ca²⁺ ions, but was not reversed when the Ca²⁺-ionophore A23187 was added to the culture media. Moreover, the effect of fosetyl-Al on peroxidase activity and peroxidase isozymes was also partially reversed by Mg²⁺ ions but not by Sr²⁺, and was accentuated by Ba²⁺ ions. These results suggested that Ca²⁺ and Mg²⁺ ions specifically overcome the inhibitory effect of fosetyl-Al on peroxidase. In this context, an apoplastic Ca²⁺/Mg²⁺-displacement hypothesis is proposed for the mechanism of action of fosetyl-Al on peroxidase from grapevine cells.     

Effects of Metal Ions on the Conformation and Activity of Acutolysin D from Agkistrodon Acutus Venom

Acutolysin D, isolated from the venom of Agkistrodon acutus, possesses marked haemorrhagic and proteolytic activities. The molecular weight and the absorption coefficients (A ^1% ₂₈₀) of acutolyisn D have been determined to be 47,850 ± 8 amu and 9.3 by mass spectrometer and UV spectrum, respectively. The effects of metal ions on the conformation and activity of acutolysin D have been studied by following fluorescence, circular dichroism and biological activity measurements. Acutolysin D contains two Ca²⁺-binding sites and two Zn²⁺-binding sites determined by atomic absorption spectrophotometer. Zn²⁺ is essential for the enzyme activities of acutolysin D, however, the presence of 1 mM Zn²⁺ significantly decreases its caseinolytic activity and intrinsic fluorescence intensity at pH 9.0 due to Zn(OH)₂ precipitate formation. Ca²⁺ is important for the structural integrity of acutolysin D, and the presence of 1 mM Ca²⁺ markedly enhances its caseinolytic activity. Interestingly, the caseinolytic activity which is inhibited partly by Cu²⁺, Co²⁺, Mn²⁺ or Tb³⁺ and inhibited completely by Cd²⁺, is enhanced by Mg²⁺. The fluorescence intensity of the protein decreases in the presence of Cu²⁺, Co²⁺, Cd²⁺ or Mn²⁺, but neither for Ca²⁺, Mg²⁺ nor for Tb³⁺. Zn²⁺, Ca²⁺, Mg²⁺, Cu²⁺, Mn²⁺, Co²⁺ and Tb³⁺ have slight effects on its secondary structure contents. In addition, Cd²⁺ causes a marked increase of antiparallel β-sheet content from 45.5% to 60.2%.     

Heavy metal ions affect the activity of DNA glycosylases of the Fpg family

Prokaryotic enzymes formamidopyrimidine-DNA glycosylase (Fpg) and endonuclease VIII (Nei) and their eukaryotic homologs NEIL1, NEIL2, and NEIL3 define the Fpg family of DNA glycosylases, which initiate the process of repair of oxidized DNA bases. The repair of oxidative DNA lesions is known to be impaired in vivo in the presence of ions of some heavy metals. We have studied the effect of salts of several alkaline earth and transition metals on the activity of Fpg-family DNA glycosylases in the reaction of excision of 5,6-dihydrouracil, a typical DNA oxidation product. The reaction catalyzed by NEIL1 was characterized by values K _m = 150 nM and k _cat = 1.2 min⁻¹, which were in the range of these constants for excision of other damaged bases by this enzyme. NEIL1 was inhibited by Al³⁺, Ni²⁺, Co²⁺, Cd²⁺, Cu²⁺, Zn²⁺, and Fe²⁺ in Tris-HCl buffer and by Cd²⁺, Zn²⁺, Cu²⁺, and Fe²⁺ in potassium phosphate buffer. Fpg and Nei, the prokaryotic homologs of NEIL1, were inhibited by the same metal ions as NEIL1. The values of I₅₀ for NEIL1 inhibition were 7 μM for Cd²⁺, 16 μM for Zn²⁺, and 400 μM for Cu²⁺. The inhibition of NEIL1 by Cd²⁺, Zn²⁺, and Cu²⁺ was at least partly due to the formation of metal-DNA complexes. In the case of Cd²⁺ and Cu²⁺, which preferentially bind to DNA bases rather than phosphates, the presence of metal ions caused the enzyme to lose the ability for preferential binding to damaged DNA. Therefore, the inhibition of NEIL1 activity in removal of oxidative lesions by heavy metal ions may be a reason for their comutagenicity under oxidative stress.     

Cd2+和Pb2+对花翅摇蚊(Chironomus kiiensis)幼虫生长发育的影响及富集效应

河流、湖泊等水生环境中普遍存在的重金属污染破坏水生生态系统并间接威胁人类健康。为探究重金属胁迫下水生昆虫花翅摇蚊（Chironomus kiiensis）生态毒理,测定了重金属Cd²⁺和Pb²⁺胁迫对花翅摇蚊化蛹率和羽化率的影响,检测了摇蚊的口器致畸与富集效应。研究结果表明,Cd²⁺和Pb²⁺影响摇蚊幼虫化蛹和羽化过程,单一重金属离子处理14 d Pb²⁺处理组的化蛹率和羽化率分别为22.22%和8.89%,低于Cd²⁺的化蛹率（25.56%）和羽化率（11.11%）,表现出更强的抑制效应。混合离子1：2和2：1配比处理组化蛹率和羽化率均为11.11%和4.44%,显著低于单一重金属离子胁迫下的化蛹率和羽化率。单一重金属离子及混合离子处理均能导致花翅摇蚊幼虫口器致畸,表现为上颚前齿断裂,中齿和基齿磨损、缺失,下唇板齿部不规则,下唇板边缘齿与中央齿磨损、断裂、增生、缺失。不同重金属离子处理下幼虫口器致畸率不同,并与暴露时间呈正相关,其中1：2配比处理14 d致畸率达到40.61%。重金属离子在摇蚊幼虫体内产生生物富集效应,单一重金属离子处理下的Pb²⁺富集含量7 d至14 d由11.46 mg/kg上升至31.32 mg/kg,不同配比混合离子处理下Pb²⁺富集含量均呈增加趋势,其中1：2配比处理组由15.48 mg/kg上升至42.50 mg/kg,而Cd²⁺在单一重金属与1：1混合离子处理组7 d至14 d的富集含量无显著性变化,2：1配比处理组由14.20 mg/kg下降至9.52 mg/kg,1：2配比由5.85 mg/kg上升至20.99 mg/kg。这些研究结果表明Cd²⁺和Pb²⁺胁迫影响花翅摇蚊幼虫生长发育且口器出现畸型,与重金属在幼虫体内的富集密切相关,为研究重金属对水生生态系统多重效应提供了理论依据。     

Effect of some heavy metal ions on copper-induced metallothionein synthesis in the yeast Saccharomyces cerevisiae

Copper-induced metallothionein (MT) synthesis in Saccharomyces cerevisiae was investigated in order to associate this exclusively with Cu²⁺ in vivo, when cultured in nutrient medium containing other heavy metal ions. Expression of the CUP1 promoter/lacZ fusion gene was inhibited by all heavy metal ions tested, especially Cd²⁺ and Mn²⁺. By adding Cd²⁺ and Mn²⁺ at 10 M concentration, the -galactosidase activity decreased by about 80% and 50% of the maximum induction observed with 1 mM CuSO₄, respectively. Furthermore, cell growth was markedly inhibited by combinations of 1 mM-Cu²⁺ and 1 M-Cd²⁺. Therefore, the yeast S. cerevisiae could not rely on MT synthesis as one of the copper-resistance mechanisms, when grown in a Cd²⁺ environment. In contrast, the presence of Mn²⁺ in the nutrient medium showed alleviation rather than growth inhibition by high concentrations of Cu²⁺. The recovery from growth inhibition by Mn²⁺ was due to decreased Cu²⁺ accumulation. Inhibitory concentrations of Co²⁺, Ni²⁺ and Zn²⁺ on expression of the CUP1p/lacZ fusion gene were at least one order of magnitude higher than that of Cd²⁺ and Mn²⁺. These results are discussed in relation to Cu²⁺ transport and Cu-induced MT synthesis in the copper-resistance mechanism of the yeast S. cerevisiae.     

嗜热四膜虫两类不同金属硫蛋白的功能补偿分析

为了分析嗜热四膜虫两类金属硫蛋白之间的关系,研究分别构建了MTT1-MTT3和MTT2-MTT4的基因敲除载体,通过同源重组获得敲除大核MTT1-MTT3和MTT2-MTT4的两种嗜热四膜虫突变体细胞株△MTT1-MTT3和△MTT2-MTT4。两种突变体细胞株暴露在Cd2+、Cu2+和H2O2的生长表现出显著不同,△MTT1-MTT3突变体细胞对Cd2+的耐受性显著下降,而△MTT2-MTT4突变体细胞对Cu2+和H2O2的耐受性均显著下降。实时荧光定量PCR分析不同突变体中其他MTT基因的表达变化,在△MTT2-MTT4突变体细胞株中,MTT5的表达水平下调,在500 mol/L Cu2+处理后,△MTT2-MTT4突变体细胞中MTT1、MTT3和MTT5表达相对野生型分别上调6.1、9.5和8.5倍。在△MTT1-MTT3突变体细胞中,MTT2、MTT4和MTT5的表达水平下调,当5 mol/L Cd2+处理后,△MTT1-MTT3突变体细胞株MTT5表达水平相对野生型上调2.9倍,而MTT2和MTT4表达水平相对野生型分别下降了4.9倍和2.5倍。结果表明嗜热四膜虫中的金属硫蛋白MTT1、MTT3和MTT5主要参与细胞的重金属解毒功能;而MTT2和MTT4主要参与细胞内正常的新陈代谢功能,不同的金属硫蛋白基因之间的表达存在相互调控和功能补偿。       

Response of Intact Cyanobacterial Cells and their Photosynthetic Apparatus to Cd2+ Ion Treatment

Intact cells of Synechococcus elongatus were treated with different concentrations (0.1 and 1.0 mM = Cd_0.1, Cd_1.0) of CdCl₂ for 24 h. Cd_0.1 treatment stimulated growth of the cell culture and chlorophyll (Chl) a concentration in the culture. Cd_1.0 inhibited both the above mentioned parameters. The oxygen evolving activity of intact cells (H₂O BQ) as well as of isolated thylakoid membranes, TM (H₂O DCPIP; H₂O PBQ + FeCy) decreased after 24 h of Cd_1.0 cultivation to 7 %. Photosystem 1 (PS1) activity was less sensitive to the effect of Cd²⁺ than PS2 activity. CdCl₂ concentration in cultivation media after 24 h of cultivation proved that the cyanobacterium cells take up these ions to a large extent from the cultivation medium. After 24 h of the Cd_1.0 treatment only 12 % of the amount of Cd²⁺ originally added to the cultivation medium was found. The ratio of external-antenna pigments, phycocyanin, and allophycocyanin to Chl increased approximately twofold with growing Cd²⁺ concentration in the cultivation medium. This ratio was found in both TM and dodecylmaltoside extracts.     

Cr3+和Cd2+对普通小球藻生长及抗氧化酶活性的影响

[目的] 为探究重金属对淡水绿藻生长的影响。[方法] 选取对水质检测具有明显指示作用的普通小球藻（Chlorella vulgaris）为实验材料,CdCl₂·2H₂O和CrCl₃·7H₂O提供重金属离子,探究不同浓度Cr³⁺和Cd²⁺在单一和复合胁迫下对藻细胞浓度、叶绿素a及相关抗氧化酶活性的影响。[结果] 随着Cr³⁺和Cd²⁺浓度不断增加,藻细胞浓度呈先增长后下降趋势;叶绿素a含量呈现先下降后升高再下降的现象,浓度为1 mg/L的单一和复合胁迫下有最大值,且毒性作用表现为Cr³⁺ < Cd²⁺ < Cr³⁺+Cd²⁺;与藻细胞膜相关的丙二醛（MDA）和过氧化氢（H₂O₂）含量随着重金属离子浓度的增大而增长;重金属离子浓度低于10 mg/L时对藻细胞内抗氧化酶系统中的超氧化物歧化酶（SOD）、过氧化氢酶（CAT）和过氧化物酶（POD）表现为促进作用,而大于10 mg/L时具有抑制作用。[结论] 结果表明在单一或复合重金属胁迫下,普通小球藻会充分调动与抗逆性相关的酶来维持自身的正常生长。     

Characterization of cadmium removal by Rhodotorula sp. Y11

Experiments were conducted studying the removal of Cd²⁺ from water via biosorption using Rhodotorula sp. Y11. The effects of temperature and initial pH of the solution on biosorption were studied. Caustic and heat treatments showed different influences on the biosorption capacity, and the highest metal uptake value (19.38 mg g⁻¹) was obtained by boiling treated yeast cells. The presence of competing cations, such as Ag⁺, Cu²⁺, and Mg²⁺, except Na⁺ ions, significantly interfered with the metal uptake. Results indicate that the Langmuir model gave a better fit to the experimental data than the Freundlich equation. The q ₁₀ value was 11.38 mg g⁻¹ for Cd²⁺ uptake by Y11. Chemical modifications of the biomass demonstrated that carboxyl and amide groups play an important role in Cd²⁺ biosorption.     

Contributions of cell wall and metal-binding peptide to Cd- and Cu-tolerances in suspension-cultured cells of tomato

Possible roles of cell wall and cytoplasmic peptides in the tolerance of cells to Cu²⁺ and Cd²⁺ ions were studied in suspension-cultured cells of tomato (Lycopersicon esculentum L. cv. Palace). Cu²⁺ and Cd²⁺ ions inhibited growth of wild type cells at concentrations more than 100 and 200 μM, respectively. Tomato cells readily developed tolerance to Cd²⁺ ions up to 1 mM but not to Cu²⁺ ions, after repeated subculturings in the presence of the respective ions. Such a metal-specific adaptation of cells was not due to the difference in the total uptakes between Cd²⁺ and Cu²⁺ ions by cells. Wild-type cells accumulated Cd²⁺ preferentially into the cytoplasmic peptide fraction and Cu²⁺ into the cell-wall fraction, when grown under the subtoxic metal conditions. Under excess metal conditions, Cd-tolerant cells produced greater amounts of Cd-binding peptides in the cytoplasm and retained lesser amounts of Cd²⁺ ions in the cell wall than did wild-type cells. In contrast, tomato cells grown in the presence of Cu²⁺ ions synthesized no detectable amounts of Cu-binding peptides in the cytoplasm and retained most of the Cu²⁺ in the cell-wall fraction, irrespective of cell lines. These results suggested that the cytoplasmic peptides rather than cell wall properties have a primary role in the response of tomato cells to excess metal environments.     

镉胁迫下硒对罗汉果组培苗光合特性的影响

实验以罗汉果组培苗为材料,室内栽培在内装市售营养土的塑料盆中,以0、10、50、100、200mg·kg-1浓度镉离子和1mg·kg-1浓度硒处理,培养20d后分析罗汉果幼苗的相关光合生理指标。结果表明:低浓度Cd2+对叶片叶绿素含量、光合速率(Pn)、蒸腾速率(Tr)、气孔导度(Gs)影响不大或稍有上升,但高浓度镉离子处理植株叶片的叶绿素含量、光合速率(Pn)、蒸腾速率(Tr)、气孔导度(Gs)明显下降;随Cd2+处理浓度的增加,叶片胞间CO2浓度(Ci)呈现上升趋势;加硒则延缓叶绿素下降,促进光合速率(Pn)、蒸腾速率(Tr)、气孔导度(Gs)上升,降低叶片胞间CO2浓度(Ci)。表明高浓度镉离子的毒害导致罗汉果组培苗叶片光合性能受到伤害,从而影响罗汉果幼苗生长。镉硒混合处理反映出硒对镉的毒害有缓解作用。     

Induction of the β-form of poly(S-carboxyethyl-l-cysteine) by divalent metalchlorides

The effects of eight divalent metal ions on fully neutralized poly(S-carboxyethyl-l-cysteine) have been studied by means of circular dichroism. Four ionic species (Cd²⁺, Cu²⁺, Zn²⁺ and Ni²⁺) effectively induce the β-form, while the other four species (Co²⁺, Ba²⁺, Ca²⁺ and Mg²⁺) are not effective. Specifically, Mg(ClO₄)₂ is ineffective, even at 1.86 m. The effect of Cu²⁺ ions on the polypeptide conformation is significant at pH values other than in the neural range. Comparison of the present results with previous ones from the lower side chain homologue, poly(S-carboxymethyl-l-cysteine), shows that Cd²⁺ and Zn²⁺ ions are more effetive but Co²⁺ ions are much less effective in the polypeptide studied here. Random coils of poly(S-carboxyethyl-l-cysteine) are more soluble while the β-form is less soluble compared with the respective conformations of the lower side-chain homologue.     

Cadmium biosorption by a cadmium resistant strain of Bacillus thuringiensis: regulation and optimization of cell surface affinity for metal cations

A marine bacterial strain putatively identified asBacillus thuringiensis strain DM55, showed multiple heavy metal resistance and biosorption phenotypes. Electron microscopic studies revealed that DM55 cells are encased in anionic cell wall polymers that can immobilize discrete aggregates of cations. Factors affecting cell surface affinity for metal cations, monitored by means of Cd²⁺ binding capability, are investigated. The mechanisms of cadmium resistance and Cd²⁺ biosorption by the bacterium appeared to be inducible and coincident. Medium components affecting metal removal under cadmium-stressed growth conditions were explored based on the application of two sequential multi-factorial statistical designs. Concentrations of potassium phosphates and peptone were the most significant variables. Optimized culture conditions allowed DM55 cells grown in the presence of 0.25 mM CdCl₂ to remove about 79% of the metal ions within 24 h with a specific biosorption capacity of 21.57 mg g^–1 of biomass. Both fresh and dry cells of DM55 prepared under cadmium-free optimal nutrient condition were also able to biosorb Cd²⁺. In addition to the concentration of phosphate in the medium, KinA, a major phosphate provider in the phosphorelay of Bacillus cells, was also demonstrated to regulate the magnitude of cell surface affinity for cadmium ions.