Inactivation and reactivation of manganese catalase: oxidation-state assignments using X-ray absorption spectroscopy.

The oxidation states of the Mn atoms in three derivatives of Mn catalase have been characterized using a combination of X-ray absorption near-edge structure (XANES) and EPR spectroscopies. The as-isolated enzyme has an average oxidation state of Mn(III) and contains a Mn(III) form, together with a reduced Mn(II) form and a variable amount (10-25%) of a Mn(III)/Mn(IV) mixed-valence derivative. Treatment with NH2OH rapidly reduces the majority of the enzyme to a Mn(II) derivative with no loss of activity. Inactivation by treatment with NH2OH + H2O2 converts all of the enzyme to a mixed-valence Mn(III)/Mn(IV) form. The inactive, mixed-valence derivative can be completely reactivated by long-term (greater than 1 h) anaerobic incubation with NH2OH, giving a reduced Mn(II)/Mn(II) derivative. These data suggest a catalytic model in which the enzyme cycles between a reduced Mn(II)/Mn(II) state and an oxidized Mn(III)/Mn(III) state.     

北方土壤中Mn的形态及其与活性Mn的关系

用逐步连续分级浸提法研究了我国北方主要土壤中Mn的存在形态及其与活性Mn的关系.碱性土壤中Mn的形态分布特征为氧化锰态Mn>残留态Mn>有机质结合态Mn>无定形铁结合态Mn>晶形铁结合态Mn>代换态Mn,与酸性土壤的排列顺序明显不同.碱性上壤条件导致土壤中的Mn更多地向生物无效态转化,使得土壤的活性Mn主要以氧化锰态和代换态存在.     

珠江三角洲典型肝癌高发区土壤锰形态及其生态效应

通过对珠江三角洲典型肝癌高发区及低发区表层、深层土壤样品和作物样品的系统采集,分别对土壤中各化学形态Mn含量及其与土壤Mn全量、土壤理化性质、蔬菜Mn富集之间的相关关系进行研究.结果表明: 研究区土壤Mn主要来自成土母质,受人类活动影响很小,肝癌高发区土壤Mn平均含量为577.65 mg·kg^-1,显著低于肝癌低发区（718.04 mg·kg^-1）和全国土壤Mn平均含量（710 mg·kg^-1）;肝癌高发区土壤Mn以残渣态和铁锰态为主,水溶态和交换态等生物有效态含量较少,二者分配系数之和不超过4%,低发区土壤Mn形态分布也有相似的分布特征,但其绝对含量显著高于肝癌高发区.Mn全量负荷水平和pH对各形态Mn的含量有重要影响,尤其是铁锰结合态、腐植酸结合态、碳酸盐态和残渣态的含量与土壤Mn全量呈显著正相关;水溶态、有机态Mn与pH呈显著负相关.5类蔬菜中,肝癌高发区油麦菜和大白菜Mn含量及富集系数显著低于肝癌低发区,其它蔬菜品种无显著差异.Mn在蔬菜中的累积量与土壤中有效态Mn（水溶态Mn与离子交换态Mn之和）呈显著正相关,而与土壤Mn全量和其他形态均无显著相关性.     

Perturbation of manganese metabolism disrupts cell division in Streptococcus pneumoniae

Manganese (Mn) is an essential micronutrient and required cofactor in bacteria. Despite its importance, excess Mn can impair bacterial growth, the mechanism of which remains largely unexplored. Here, we show that proper Mn homeostasis is critical for cellular growth of the major human respiratory pathogen Streptococcus pneumoniae. Perturbations in Mn homeostasis genes, psaBCA, encoding the Mn importer, and mntE, encoding the Mn exporter, lead to Mn sensitivity during aerobiosis. Mn‐stressed cells accumulate iron and copper, in addition to Mn. Impaired growth is a direct result of Mn toxicity and does not result from iron‐mediated Fenton chemistry, since cells remain sensitive to Mn during anaerobiosis or when hydrogen peroxide biogenesis is significantly reduced. Mn‐stressed cells are significantly elongated, whereas Mn‐limitation imposed by zinc addition leads to cell shortening. We show that Mn accumulation promotes aberrant dephosphorylation of cell division proteins via hyperactivation of the Mn‐dependent protein phosphatase PhpP, a key enzyme involved in the regulation of cell division. We discuss a mechanism by which cellular Mn:Zn ratios dictate PhpP specific activity thereby regulating pneumococcal cell division. We propose that Mn‐metalloenzymes are particularly susceptible to hyperactivation or mismetallation, suggesting the need for exquisite cellular control of Mn‐dependent metabolic processes.     

Analysis of whole blood manganese by flameless atomic absorption spectrophotometry and its use as an indicator of manganese status in animals

To investigate the use of whole blood manganese (Mn) as an indicator of total body Mn, we measured Mn in whole blood and liver of rats fed purified diets containing adequate (45 micrograms Mn/g diet) or deficient (1 microgram Mn/g diet) Mn. The mean hepatic Mn concentration was significantly lower (P less than 0.001) in the Mn-deficient group compared to the control group, 0.36, microgram Mn/g and 1.73 micrograms Mn/g, respectively. Furthermore, whole blood Mn was significantly reduced (P less than 0.001) in the deficient group when compared to the control group, 4.0 ng Mn/ml and 8.6 ng Mn/ml, respectively. Hepatic Mn linearly regressed against whole blood Mn yielded a statistically significant (P less than 0.001) correlation coefficient of 0.775. These data suggest that whole blood Mn is a valid indicator of body Mn status and thus may be useful, in addition to the measurements of serum copper and zinc, for the diagnosis and prognosis of diseases in which the metabolism of trace elements is affected. In addition, this paper describes and delineates operational parameters for the measurement of whole blood Mn using the IL 551 atomic absorption spectrophotometer and the IL 555 B flameless atomizer.     

Influence of manganese deficiency and toxicity on isoprenoid syntheses

Twenty-eight day old wheat (Triticum aestivum L. cv Stacy) response to varying Mn concentration (10.1-10,000 micromolar) in nutrient solution was measured. Manganese concentrations in the most recently matured leaves (blade 1) were 0.21 to 19.03 mmol Mn per kilogram dry weight, respectively. Fresh and dry weights increased to a maximum at the 5 micromolar Mn nutritional level (0.37 millimole Mn per kilogram dry weight) and were decreased at Mn above and below this concentration. Blade 1 chloroplast pigment concentrations increased up to the 20 micromolar Mn nutritional level (1.98 millimole Mn per kilogram dry weight) and decreased at higher Mn concentrations. Thylakoid Mn content was above 1 mole Mn/100 mole chloroplast at Mn nutrition levels which resulted in greatly decreased plant growth. Total phytoene biosynthesis was decreased by Mn deficiency and toxicity. In vitro ent- kaurene synthesis was greatly influenced by Mn concentration with a maximal biosynthesis at 1 micromolar Mn and decreases at Mn levels above and below this concentration. In vivo blade 1 gibberellic acid equivalent concentrations were maximal at 20 parts per million Mn nutrition solution levels (1.98 millimole Mn per kilogram dry weight) and decreased at Mn tissue concentrations above and below this value; additionally, gibberellic acid concentrations were reciprocal to extracted C₂₀ alcohol concentrations. Mn influence on gibberellin and chloroplast pigment biosyntheses exactly matched the measured changes in growth.     

Genotypic differences in manganese efficiency: field experiments with␣winter barley (Hordeum vulgare L.)

Eight different winter barley genotypes were grown in a plot experiment in northern Denmark, on soil where Mn deficiency had been previously demonstrated, to assess field performance and grain yield under two treatments: with foliar Mn fertilisation and without foliar Mn fertilisation. A high Mn efficiency was demonstrated for two of the genotypes and a low Mn efficiency was observed for one of the genotypes in three consecutive years; highly variable results were obtained for other genotypes. Mn efficiency based on grain yields could not be related to whole shoot Mn concentration, biomass or Mn uptake at various vegetative growth stages. Chlorophyll a fluorescence analysis was able to differentiate between the two Mn treatments up to 7weeks after the last Mn application, whereas Mn concentrations, shoot biomass and Mn accumulation could not. It was not possible to fully alleviate Mn deficiency by repeated foliar spraying on the control plots. Given the growing conditions at the trial site and the low Mn in plant tissue, it is possible that Mn deficiency was extreme enough that Mn efficiency mechanisms broke down.     

Estimating daily intakes of manganese due to breast milk,infant formulas,or young child nutritional beverages in the United States and France: Comparison to sufficiency and toxicity thresholds

BackgroundAlthough manganese (Mn) is an essential nutrient, recent research has revealed that excess Mn in early childhood may have adverse effects on neurodevelopment.MethodsWe estimated daily total Mn intake due to breast milk at average body weights by reviewing reported concentrations of breast milk Mn and measurements of body weight and breast milk intake at 3 weeks, 4.25 months, 7 months, and 18 months. We compared these figures to the Mn content measured in 44 infant, follow-up, and toddler formulas purchased in the United States and France. We calculated Mn content of formula products made with ultra-trace elemental analysis grade water (0 μg Mn/L) and with water containing 250 μg Mn/L, a concentration which is relatively high but less than the World Health Organization Health-based value of 400 μg Mn/L or the United States Environmental Protection Agency Health Advisory of 350 μg Mn/L.ResultsEstimated mean daily Mn intake from breast milk ranged from 1.2 μg Mn/kg/day (3 weeks) to 0.16 μg Mn/kg/day (18 months), with the highest intakes at the youngest age stage we considered, 3 weeks. Estimated daily Mn intake from formula products reconstituted with 0 μg Mn/L water ranged from 130 μg Mn/kg/day (3 weeks) to 4.8 μg Mn/kg/day (18 months) with the highest intakes at 3 weeks. Formula products provided 28–520 times greater than the mean daily intake of Mn from breast milk for the 4 age stages that we considered. Estimated daily Mn intake from formula products reconstituted with water containing 250 μg Mn/L ranged from 12 μg Mn/kg/day to 170 μg Mn/kg/day, which exceeds the United States Environmental Protection Agency Reference Dose of 140 μg Mn/kg/day for adults.ConclusionsMn deficiency is highly unlikely with exclusive breast milk or infant formula feeding, but established tolerable daily intake levels for Mn may be surpassed by some of these products when following labeled instructions.     

Effect of manganese spatial distribution in the soil profile on wheat growth in rice–wheat rotation

Manganese (Mn) deficiency in wheat under rice (Oryza sativa L.) and wheat (Triticum aestivum L.) rotation is an important problem in most rice-growing areas in China. A field survey, field trials and a soil column experiment were conducted to determine the relationship between Mn leaching and distribution in soil profiles and paddy rice cultivation and the effects of Mn distribution in soil profiles on wheat growth and its response to Mn fertilization. At five field sites surveyed, total Mn and active Mn concentrations in the topsoil layers under rice–wheat rotations were only 42% and 11%, respectively, of those under systems without paddy rice. Both total and available Mn increased with soil depth in soils with rice–wheat rotations, showing significant spatial variability of Mn in the soil profile. Manganese leaching was the main pathway for Mn loss in coarse-textured soil with high pH, while excessive Mn uptake was the main pathway for Mn loss in clay-textured and acid soil. When Mn was deficient in the topsoil, sufficient Mn in the subsoil contributed to better growth and Mn nutrition of wheat but insufficient Mn in the subsoil resulted in Mn deficiency in wheat.     

Residual Mn and associated soybean yield response from MnSO4 application on a sandy loam soil

Summary A field experiment was initiated in 1977 to evaluate soybean (Glycine max (L.) Merr.) response to rates of broadcast Mn and was continued in 1978 to determine the residual effects of applied Mn. This study was conducted on a Dragston fine sandy loam soil with an initial pH of 6.0 and 2.5% organic matter. In 1977 the 40kg Mn/ha rate increased seed yields by 900 kg/ha and raised leaf tissue Mn concentration from 10.3 to 16.1 g Mn/g. Residual Mn increased seed yield and leaf Mn concentration. Reapplied Mn produced higher yields and tissue Mn levels than the residual Mn. The Mitscherlich plant growth model r=0.67^**, fit the relationship between yield and leaf Mn concentration. Double acid extractable Mn in soil sampled in 1978 was linearly related to broadcast Mn and soybean yields. Applied Mn increased Mn concentrations of soybean seed parts by 97%, 182% and 109% in the cotyledon, embryo and seedcoat, respectively.     

Factors responsible for genotypic manganese tolerance in cowpea (Vigna unguiculata)

In experiments with 29 cowpea genotypes considerable variation in Mn tolerance could be found. Ranking according to Mn tolerance was almost the same in sand and water culture. Mn tolerance is not related to greater vigour or exclusion of Mn from uptake and translocation, but depends mainly on the internal tolerance to excess Mn especially in the leaf tissue.Growth depression by Mn excess is characterized by local accumulatiòn of Mn, deposition of Mn oxides, and typical macro-symptoms on the older leaves (brown spotsclorosisshedding of the leaves). Autoradiographic studies with⁵⁴Mn and extraction of the leaves with methanol and H₂O indicate a causal relationship between Mn tolerance and the more homogenous distribution of Mn in the tissue. In tolerant genotypes local accumulation and deposition of Mn is inhibited or retarded.Mn applied to the petioles of fully expanded leaves induces the same toxicity symptoms on the leaf blades as Mn absorbed by the roots. There is a good agreement between the rankings of the different genotypes for Mn tolerance according to the depression of shoot dry matter production by Mn excess in long term pot experiments and the appearance of toxicity symptoms after application of Mn to the petioles.The regulation of Mn tolerance at the leaf tissue level allows a quick and non-destructive screening of large numbers of genotypes for Mn tolerance.     

Manganese (Mn) Oxidation Increases Intracellular Mn in Pseudomonas putida GB-1

Bacterial manganese (Mn) oxidation plays an important role in the global biogeochemical cycling of Mn and other compounds, and the diversity and prevalence of Mn oxidizers have been well established. Despite many hypotheses of why these bacteria may oxidize Mn, the physiological reasons remain elusive. Intracellular Mn levels were determined for Pseudomonas putida GB-1 grown in the presence or absence of Mn by inductively coupled plasma mass spectrometry (ICP-MS). Mn oxidizing wild type P. putida GB-1 had higher intracellular Mn than non Mn oxidizing mutants grown under the same conditions. P. putida GB-1 had a 5 fold increase in intracellular Mn compared to the non Mn oxidizing mutant P. putida GB-1-007 and a 59 fold increase in intracellular Mn compared to P. putida GB-1 ∆2665 ∆2447. The intracellular Mn is primarily associated with the less than 3 kDa fraction, suggesting it is not bound to protein. Protein oxidation levels in Mn oxidizing and non oxidizing cultures were relatively similar, yet Mn oxidation did increase survival of P. putida GB-1 when oxidatively stressed. This study is the first to link Mn oxidation to Mn homeostasis and oxidative stress protection.     

Chloramphenicol-inducible gene expression in Bacillus subtilis is independent of the chloramphenicol acetyltransferase structural gene and its promoter

Lactobacillus plantarum has an unusually high Mn(II) requirement for growth and accumulated over 30 mM intracellular Mn(II). The acquisition of Mn(II) by L. plantarum occurred via a specific active transport system powered by the transmembrane proton gradient. The Mn(II) uptake system has a Km of 0.2 microM and a Vmax of 24 nmol mg-1 of protein min-1. Above a medium Mn(II) concentration of 200 microM, the intracellular Mn(II) level was independent of the medium Mn(II) and unresponsive to oxygen stresses but was reduced by phosphate limitation. At a pH of 5.5, citrate, isocitrate, and cis-aconitate effectively promoted MN(II) uptake, although measurable levels of 1,5-[14C]citrate were not accumulated. When cells were presented with equimolar Mn(II) and Cd(II), Cd(II) was preferentially taken up by the Mn(II) transport system. Both Mn(II) and Cd(II) uptake were greatly increased by Mn(II) starvation. Mn(II) uptake by Mn(II)-starved cells was subject to a negative feedback regulatory mechanism functioning less than 1 min after exposure of the cells to Mn(II) and independent of protein synthesis. When presented with a relatively large amount of exogenous Mn(II), Mn(II)-starved cells exhibited a measurable efflux of their internal Mn(II), but the rate was only a small fraction of the maximal Mn(II) uptake rate.     

Effects of copper, iron, and ascorbic acid on manganese availability to rats.

Four experiments were done to characterize the interactions of copper, iron, and ascorbic acid with manganese in rats. All experiments were factorially arranged Dietary Mn concentrations were less than 1 micrograms/g (Mn0) and 50 micrograms/g (Mn+). Dietary Cu was less than 1 mg/g (Cu0) and 5 micrograms/g (Cu+); dietary Fe was 10 micrograms/g (Fe10) and 140 micrograms/g (Fe140). Ascorbic acid (Asc) was not added to the diet or added at a concentration of 10 g/kg diet. Experiment 1 had two variables, Mn and Cu; in Experiment 2, the variables were Mn and Asc. In Experiment 3, the variables were Mn, Cu, and Asc; in Experiment 4, they were Mn, Cu, and Fe. Definite interactions between Mn and Cu were observed, but they tended to be less pronounced than interactions between Mn and Fe. Cu depressed absorption of 54Mn and accelerated its turnover. In addition, adequate Cu (Cu+), compared with Cu0, depressed liver, plasma, and whole blood Mn of rats. Absorption of 67Cu was higher in animals fed Mn0 diets than in those fed Mn+. Ascorbic acid depressed Mn superoxide dismutase activity and increased Cu superoxide dismutase activity in the heart. The addition of ascorbic acid to the diet did not affect Mn concentration in the liver or blood. Absorption of 54Mn was depressed in rats fed Fe140 compared with those fed Fe10. Interactions among Fe, Cu, and Mn resulted in a tendency for Mn superoxide dismutase activity to be lower in rats fed Fe140 than in rats fed Fe10. Within the physiologic range of dietary concentrations, Mn and Cu have opposite effects on many factors that tend to balance one another. The effects of ascorbic acid on Mn metabolism are much less pronounced than effects of dietary Cu, which in turn affects Mn metabolism less than does Fe.     

超富集植物短毛蓼对锰的富集特征

通过野外调查和营养液培养试验,研究了锰在短毛蓼体内的富集特征和对其生长的影响.在锰含量高达2.5×105mg/kg的锰矿废弃地上短毛蓼生长良好,叶锰含量高达1.66×104mg/kg.营养液培养条件下,随着生长介质中Mn浓度的升高,短毛蓼根、茎、叶中的Mn含量逐渐增加,当锰供应水平为1.000mmol/L时,叶锰含量超过10000mg/kg;当锰供应水平为20 000mmol/L时,短毛蓼仍能生长,根、茎和叶3部分的锰含量均达到最大值,分别为9923,18112mg/kg和55750mg/kg.在所有锰供应水平下,短毛蓼茎和叶中的锰含量都比根部的高.结果表明,短毛蓼是一种锰超富集植物,这一发现为锰污染土壤的植物修复和探讨锰在植物体内的超富集机理提供了一种新的种质资源.     

环境条件对不同品种小麦缺Mn的影响

在温室和网室中进行盆栽试验 ,研究不同环境条件和杂草对冬小麦缺Mn的影响 ,同时探讨不同小麦品种对缺Mn的耐性 ,发现生长在温室中的小麦其缺Mn症尤为严重 ,而从网室转移到温室的小麦因苗期在网室中受降雨渍水影响吸收了较多Mn2 ,缺Mn症就较轻 ,表明环境因子中降雨是影响小麦Mn营养的一个重要因素 ;供试的 3个品种中 331 7较耐缺Mn ,其Mn的吸收量明显高于敏感品种 .此外 ,试验还发现与小麦伴生的杂草麦麦草对Mn吸收能力强 ,是小麦根际Mn营养的有力争夺者 .     

土壤中铁锰的形态分布及其有效性研究

以乌鲁木齐雅马里克山的土壤为研究对象,采用Tessier连续提取法对土壤铁、锰各种化学形态进行浸提.研究了土壤中有效铁、锰和土壤的理化性质与土壤铁、锰形态之间的关系,及对铁、锰在土壤中存在形态的影响;并通过盆栽试验对铁、锰的植物有效性进行了分析.结果表明,土壤铁主要以残渣态为主,占全铁的92.3%,其它形态含量均小于全量的8%.土壤锰主要是以铁锰氧化态和残渣态为主,分别占全锰的49%和41.6%,其它形态含量均小于全量的10%.用二级出水灌溉处理可增加铁、锰的有效性,原污水灌溉不利于铁、锰的供应,土壤缺铁、锰的现象可通过施加一定量的铁盐和锰盐而得以改善.相关分析还表明,土壤的理化性质与铁锰形态之间有一定的相关性.供试土壤的pH值、CaCO3含量、有机质及阳离子交换态等对土壤铁、锰的有效性影响较大.逐步回归分析表明,铁的氧化物结合态对植物最为有效,锰的有机结合态对植物有效性贡献最大.     

Redistribution of Manganese in Maturing Lupin us angustifolius cv. Illyarrie in Relation to Levels of Previous Accumulation

The retranslocation of Mn from pools of pre-accumulation todeveloping seed of Lupinus angustifolius was examined in growthchamber and field experiments. In the growth chamber experiment, plants (cv. Illyarrie) weregrown in sand containing Mn at 18 levels of supply from low(15 fig Mn kg"1) to luxury (12800 fig Mn kg{small tilde}l).At 90 d, sand was washed from the roots and the plants weregrown to maturity in nutrient solution in the absence of Mn.At sequential harvests, the plants were divided into a numberof fractions and were analysed for Mn to determine changes inthe amount of Mn contained in each component. Plants (cv. Marri)were also grown in the field at a Mn deficient site with andwithout added Mn, and similar harvests and observations weremade. Manganese moved readily from roots, stems and petioles to developingsinks, including seeds via pods. Retranslocation of Mn preventedthe appearance of Mn deficient seed (‘split seed’)in plants which had accumulated sufficient Mn prior to the omissionof Mn supply. The mode of Mn transport is discussed, and itsconveyance to seed in phloem, mediated by transfer cells, issuggested. The identification of pools of Mn accumulation fromwhich Mn may retranslocate presents a basis on which tests prognosticof impending Mn deficiency in maturing lupins may be developed.Stem is favoured as the tissue on which to base such tests. No Mn translocated to developing seed sinks from mature, oreven senescing lupin leaves. The cause appears to be that Mnis poorly loaded into the phloem of leaves although much wasfound to be leachable. About 40 per cent of leaf Mn was extractablewith water, more (approx. 70 per cent) with dilute ethylenediamine tetraacen’c acid but little (approx. 14 per cent)with N7>Jdimethyl formamide along with other cell contents.The form of Mn in leaves is discussed.     

Accumulation and detoxification of manganese in hyperaccumulator Phytolacca americana

Pokeweed ( Phytolacca americana ) has recently received much attention because of its ability to hyperaccumulate manganese (Mn). The internal mechanism of detoxification of Mn, however, is not fully understood. In the present study, we investigated Mn accumulation, subcellular distribution, chemical speciation and detoxification through oxalate in pokeweed. The plant accumulated excess Mn in the leaves, mainly in the water-soluble fraction, and over 80% of Mn was in a water-soluble form, while accumulation of excess Mn in the cellular organelle and membrane fraction caused phytotoxicity. In addition, pokeweed has an intrinsically high oxalate content. In all experiments, there was sufficient oxalate to chelate Mn in leaf water extracts at all different levels of Mn application. Phase analysis of X-ray diffraction detected oxalate–Mn chelate complexes, and gel chromatography further confirmed the chelation of Mn by oxalate. In conclusion, pokeweed accumulates excess Mn in the soluble fraction of leaf cells, most likely in vacuoles, in which detoxification of Mn could be achieved by chelation with oxalate.