一株氧化亚铁硫杆菌的筛选及对低品位碲矿的浸出

从四川石棉矿区的酸性矿坑水和土壤中筛选到一株细菌PD-1,对其形态、理化特征及16SrDNA序列研究表明:细菌细胞呈杆状,革兰氏染色阴性,最适生长温度30℃,最适初始pH2.0,其16SrDNA序列与氧化亚铁硫杆菌的序列相似度高达99%以上,可鉴定为氧化亚铁硫杆菌菌株。利用该菌株对低品位碲矿进行摇瓶浸出实验,结果表明:菌株在接种量10%、温度30℃、初始pH2.0、转速150r/min、碲矿粒度-150μm及矿浆浓度2%的条件下浸矿,30d碲的浸出率可达67.8%。     

一株氧化亚铁硫杆菌的筛选及对低品位碲矿的浸出北大核心

从四川石棉矿区的酸性矿坑水和土壤中筛选到一株细菌PD-1,对其形态、理化特征及16SrDNA序列研究表明:细菌细胞呈杆状,革兰氏染色阴性,最适生长温度30℃,最适初始pH2.0,其16SrDNA序列与氧化亚铁硫杆菌的序列相似度高达99%以上,可鉴定为氧化亚铁硫杆菌菌株。利用该菌株对低品位碲矿进行摇瓶浸出实验,结果表明:菌株在接种量10%、温度30℃、初始pH2.0、转速150r/min、碲矿粒度-150μm及矿浆浓度2%的条件下浸矿,30d碲的浸出率可达67.8%。     

耐高矿化度浸矿菌的驯化及浸铀效果

从721铀矿石样中分离出的一株氧化亚铁钩端螺旋菌(Leptospirillums ferrooxidans)与氧化亚铁硫杆菌(Thiobacillus ferrooxidans)的混合菌株,经不同条件的驯化及紫外线诱变处理后,可在高矿化度(M>50g/L)、低pH(pH=1.4)的铀矿石酸化液中良好生长。实验室槽浸实验表明,这一混合菌株对低pH、高矿化度的浸出体系具有良好的适应性,仅用7天时间,渣计浸出率达90.06%,浸出过程中未出现铁损失。     

耐低pH氧化亚铁硫杆菌的培养与驯化

采用9K培养基从沙洲坝铀矿石中分离出一株耐低pH(pH=1.6)的氧化亚铁硫杆菌(B16),经多次驯化,它可以适应矿石培养基和矿石酸化液。从而证实,用矿石酸化液替代9K培养基培养氧化亚铁硫杆菌是可行的。     

不同初始条件对细菌浸出电子线路板中铜的影响

利用筛选的氧化亚铁硫杆菌,研究了初始pH、Fe2+质量浓度以及氧化亚铁硫杆菌的驯化对电子线路板中金属铜浸出的影响。结果表明:酸性条件有利于保持铁离子浓度,从而提高浸出速度,浸出最佳pH为1.5;初始Fe2+质量浓度越高,越有利于铜的浸出,最佳Fe2+质量浓度为9.0 g/L;氧化亚铁硫杆菌驯化与否对铜的浸出影响不大。     

嗜酸性氧化亚铁硫杆菌对四川某铜矿选冶渣重金属的阻滞

为修复治理四川某选冶渣重金属污染,降低选冶渣中多重金属生物有效性,以嗜酸性氧化亚铁硫杆菌为菌种、四川某选冶渣为研究对象,研究嗜酸性氧化亚铁硫杆菌生长特性及其对选冶渣中多种重金属的阻滞效果。结果表明,当初始pH=2.0、接种量15%时,选冶渣中Fe、Zn、Cu、Pb生物有效性降低率分别为98.05%、0、0、100%;当初始pH=5.0、接种量5%时,选冶渣中Fe、Zn、Cu、Pb生物有效性降低率分别为93.03%、62.92%、56.44%、52.56%。采用嗜酸性氧化亚铁硫杆菌滞固该铜选冶矿中多重金属具有可行性,可为同类选冶渣修复治理提供借鉴。     

JX嗜酸异养菌与氧化亚铁硫杆菌联合浸铀的研究

从江西某铀矿酸性矿坑水中筛选出了一株JX嗜酸异养菌和一株氧化亚铁硫杆菌,将它们按不同的接种比例和接种时间接种于加了铀矿粉的9K培养基中,进行了联合浸铀试验。结果表明:加入异养菌后,浸出体系中Fe2+氧化速率、pH下降速度及Eh的上升速度都有所减慢;不同接种比例试验的铀的最终浸出率均在97%左右;不同接种时间试验的铀最终浸出率均在98%左右;JX异养菌的加入,对氧化亚铁硫杆菌浸铀没有起到明显的促进作用。     

氧化亚铁硫杆菌最佳生长条件的初步探索

氧化亚铁硫杆菌生长活性影响因素有很多，其中主要的是温度、pH、培养基的量以及接种量等。用比浊法以及二价铁离子的变化为指标，对氧化亚铁硫杆菌的生长条件进行初步研究，得出其最佳生长条件为：温度28℃左右，初始pH为2．3左右，培养基装量为50～100mL／250mL。接种量为10％。     

氧化亚铁硫杆菌柱浸脱硫试验

采用氧化亚铁硫杆菌（简称T．f．菌）脱除内蒙古平庄燃煤中硫，研究了柱浸脱硫过程中影响脱硫率的各种影响因素及脱硫效果。进行了煤粒度比较试验、氧化亚铁硫杆菌不同接种量比较试验、菌液酸度比较试验、氮磷源比较试验。结果表明，脱硫效率与煤的粒度、接种量、菌液pH值、氮磷源加放次序、上柱循环时间有关；在pH值2．0～2．5，接种量15％，一次加齐氮磷源，煤的粒度为小于88mm的条件下脱硫率最好；在上述条件下细菌脱硫后的含硫量大大降低，脱硫率可达42．1％。     

氧化亚铁硫杆菌亚铁氧化活性诱变育种理论探讨

结合已有的实验研究探讨了氧化亚铁硫杆菌亚铁氧化活性诱变育种过程中出现的几个理论问题 :突变基因性质 ;正突变与细菌修复系统 ;诱变剂选择与“钝化”防止 ;遗传稳定性与抑制基因突变 ;有效的育种程序。以期在进一步诱变育种中加快进程 ,获得遗传稳定、氧化快的突变株     

Crystallographic and mutational analyses of an extremely acidophilic and acid-stable xylanase: biased distribution of acidic residues and importance of Asp37 for catalysis at low pH

Xylanase C from Aspergillus kawachii has an optimum pH of 2.0 and is stable at pH 1.0. The crystal structure of xylanase C was determined at 2.0 A resolution (R-factor = 19.4%). The overall structure was similar to those of other family 11 xylanases. Asp37 and an acid-base catalyst, Glu170, are located at a hydrogen-bonding distance (2.8 A), as in other xylanases with low pH optima. Asp37 of xylanase C was replaced with asparagine and other residues by site-directed mutagenesis. Analyses of the wild-type and mutant enzymes showed that Asp37 is important for high enzyme activity at low pH. In the case of the asparagine mutant, the optimum pH shifted to 5.0 and the maximum specific activity decreased to about 15% of that of the wild-type enzyme. On structural comparison with xylanases with higher pH optima, another striking feature of the xylanase C structure was found; the enzyme has numerous acidic residues concentrated on the surface (so-called 'Ser/Thr surface' in most family 11 xylanases). The relationship of the stability against extreme pH conditions and high salt concentrations with the spatially biased distribution of charged residues on the proteins is discussed.     

Optimisation of different physical parameters for bioleaching of phosphate by Aspergillus niger from Indian rock phosphate

A mutant strain of Aspergillus niger AB100 was incubated with samples of rock phosphate. Mutation resulted in a greater amount of solubilisation (30 to 35%) as against the parent strain (10 to 15%). The influence of leaching parameters such as ore concentration (pulp density), particle size, initial pH of the medium, temperature, volume of the medium in 250 ml flasks, inoculum concentration and age of inoculum was studied. When low quantity of rock phosphate is applied (0.1%) the solubilisation of phosphorus was optimal (40.5%). Optimum particle size was--200 to 240 mesh, initial pH of the medium 4.0, optimum volume of the fermentation medium 160 ml, time period of incubation was 8 days, inoculum volume was 7.5 ml, and age of inoculum 7 days. The maximum leaching of phosphorus by using these optimum physical parameters is 45 to 50%.     

Effects of changes in strain path on work-hardening in CP aluminium and an AlCuMg alloy

Effects of abrupt changes in the direction of plastic deformation on work-hardening behaviour have been investigated in two-stage stretching of sheets of AA1050 and a heat-treated 2014 aluminium alloy aged at various temperatures up to 300°C. The results show that reorganisation of dislocation distribution after a change in strain path can result in transient changes in work-hardening behaviour of two kinds. Changes of the first kind, which tend to increase the hardening rate in early stages of the second mode of deformation, are associated with reorientation of internal stresses. Changes in the second kind, which tend to cause transient reductions in hardening rate, are believed to be associated with partial dissolution of the original dislocation substructure. The relative magnitudes and strain dependencies of these two kinds of change depend on the deformation sequence and on material variables. The change in hardening rate of CP aluminium after a change in strain path is dominated by changes of the second kind which, after moderate prestrains, cause reductions in the limits of stable elongation. In similar tests on overaged conditions of the 2014 alloy the overall changes in hardening rate are dominated by changes of the first kind, so that the limit of uniform elongation is increased by a change in strain path. When dynamic ageing is active in the 2014 alloy changes of the second kind can be suppressed so that reductions in the hardening rate do not occur.     

A triple mutation in the a subunit of the Escherichia coli/Propionigenium modestum F1Fo ATPase hybrid causes a switch from Na+ stimulation to Na+ inhibition

Previously we have shown that the Na+-translocating Escherichia coli (F1-delta)/Propionigenium modestum (Fo+delta) hybrid ATPase acquires a Na+-independent phenotype by the c subunit double mutation F84L, L87V that is reflected by Na+-independent growth of the mutant strain MPC8487 on succinate [Kaim, G., and Dimroth, P. (1995) J. Mol. Biol. 253, 726-738]. Here we describe a new class of mutants that were obtained by random mutagenesis and screening for Na+-independent growth on succinate. All six mutants of the new class contained four mutations in the a subunit (S89P, K220R, V264E, I278N). Results from site-specific mutagenesis revealed that the substitutions K220R, V264E, and I278N were sufficient to create the new phenotype. The resulting E. coli mutant strain MPA762 could only grow in the absence but not in the presence of Na+ ions on succinate minimal medium. This effect of Na+ ions on growth correlated with a Na+-specific inhibition of the mutant ATPase. The Ki for NaCl was 1. 5 mM at pH 6.5, similar to the Km for NaCl in activating the parent hybrid ATPase at this pH. On the other hand, activation by Li+ ions was retained in the new mutant ATPase. In the absence of Na+ or Li+, the mutant enzyme had the same pH optimum at pH 6.5 and twice the specific activity as the parent hybrid ATPase. In accordance with the kinetic data, the reconstituted mutant ATPase catalyzed H+ or Li+ transport but no Na+ transport. These results show for the first time that the coupling ion selectivity of F1Fo ATPases is determined by structural elements not only of the c subunit but also of the a subunit.     

Bacterial proteases: production, isolation and utilization in animal nutrition

Under conditions of submerged fermentation of Bacillus licheniformis strain L-3 in 15-L MBR-Schulzer bioreactor, the maximum production of proteolytic enzymes was achieved in the nutrient medium which contained 1% milk powder, 0.3% yeast autolysate, 0.5% corn starch and malt (20 ml per 100 ml of the medium). The preparation obtained of extracellular alkaline bacterial Ser-protease of the subtilisin type is characterized by optimum pH 9.8-10.2 good up to a temperature stability (65 degrees C) and has molecular weight cca of 26 kDa. The use of chemical mutagens (HNO2, 5-bromouracil) has enabled to select new strains (L-3N and L-3U) whose protease activity is 1.8-2.2 times higher as compared to the original Bacillus licheniformis strain L-3. The addition of these enzymes to the fodder has a positive effect on the retention of nitrogen substances.     

PHR2 of Candida albicans encodes a functional homolog of the pH-regulated gene PHR1 with an inverted pattern of pH-dependent expression

Deletion of PHR1, a pH-regulated gene of Candida albicans, results in pH-conditional defects in growth, morphogenesis, and virulence evident at neutral to alkaline pH but absent at acidic pH. Consequently, we searched for a functional homolog of PHR1 active at low pH. This resulted in the isolation of a second pH-regulated gene, designated PHR2. The expression of PHR2 was inversely related to that of PHR1, being repressed at pH values above 6 and progressively induced at more acidic pH values. The predicted amino acid sequence of the PHR2 protein, Phr2p, was 54% identical to that of Phr1p. A PHR2 null mutant exhibited pH-conditional defects in growth and morphogenesis analogous to those of PHR1 mutants but manifest at acid rather than alkaline pH values. Engineered expression of PHR1 at acid pH in a PHR2 mutant strain and PHR2 at alkaline pH in a PHR1 mutant strain complemented the defects in the opposing mutant. Deletion of both PHR1 and PHR2 resulted in a strain with pH-independent, constitutive growth and morphological defects. These results indicate that PHR1 and PHR2 represent a novel pH-balanced system of functional homologs required for C. albicans to adapt to environments of diverse pH.     

Proton gradient-driven nickel uptake by vacuolar membrane vesicles of Saccharomyces cerevisiae

A vacuolar H+-ATPase-negative mutant of Saccharomyces cerevisiae was highly sensitive to nickel ion. Accumulation of nickel ion in the cells of this mutant of less than 60% of the value for the parent strain arrested growth, suggesting a role for this ATPase in sequestering nickel ion into vacuoles. An artificially imposed pH gradient (interior acid) induced transient nickel ion uptake by vacuolar membrane vesicles, which was inhibited by collapse of the pH difference but not of the membrane potential. Nickel ion transport into vacuoles in a pH gradient-dependent manner is thus important for its detoxification in yeast.     

Gap junctional communication and the tyrosine phosphorylation of connexin 43 in interaction between breast cancer and endothelial cells

An antigenic double mutant of rabies virus (challenge virus standard [CVS] strain) was selected by successive use of two neutralizing antiglycoprotein monoclonal antibodies, both specific for antigenic site III. This mutant differed from the original virus strain by two amino acid substitutions in the ectodomain of the glycoprotein. The lysine in position 330 and the arginine in position 333 were replaced by asparagine and methionine, respectively. This double mutant was not pathogenic for adult mice. When injected intramuscularly into the forelimbs of adult mice, this virus could not penetrate the nervous system, either by the motor or by the sensory route, while respective single mutants infected motoneurons in the spinal cord and sensory neurons in the dorsal root ganglia. In vitro experiments showed that the double mutant was able to infect BHK cells, neuroblastoma cells, and freshly prepared embryonic motoneurons, albeit with a lower efficiency than the CVS strain. Upon further incubation at 37 degrees C, the motoneurons became resistant to infection by the mutant while remaining permissive to CVS infection. These results suggest that rabies virus uses different types of receptors: a molecule which is ubiquitously expressed at the surface of continuous cell lines and which is recognized by both CVS and the double mutant and a neuron-specific molecule which is not recognized by the double mutant.     

The Feigenbaum scenario as a model of the limits of conscious information processing

Helicobacter pylori persists in the human stomach where it may encounter a variety of DNA-damaging conditions, including gastric acidity. To determine whether the nucleotide excision repair (NER) pathway contributes to the repair of acid-induced DNA damage, we have cloned the putative H. pylori NER gene, uvrB. Degenerate oligonucleotide primers based on conserved amino acid residues of bacterial UvrB proteins were used in PCR with genomic DNA from H. pylori strain 84-183, and the 1.3-kb PCR product from this reaction was used as a probe to clone uvrB from an H. pylori genomic library. This plasmid clone had a 5.5-kb insert containing a 2.0-kb ORF whose predicted product (658 amino acids; 75.9 kDa) exhibited 69.5% similarity to E. coli UvrB. We constructed an isogenic H. pylori uvrB mutant by inserting a kanamycin-resistance cassette into uvrB and verified its proper placement by Southern hybridization. As with uvrB mutants of other bacteria, the H. pylori uvrB mutant showed a greatly increased sensitivity to the DNA-damaging agents methylmethane sulfonate and ultraviolet radiation. The uvrB mutant also was significantly more sensitive than the wild-type strain to killing by low pH, suggesting that the H. pylori nucleotide excision repair (NER) pathway is involved in the repair of acid-induced DNA damage.     

Photosynthetic electron transport involved in PxcA-dependent proton extrusion in Synechocystis sp. Strain PCC6803: effect of pxcA inactivation on CO2, HCO3-, and NO3- uptake

The product of pxcA (formerly known as cotA) is involved in light-induced Na+-dependent proton extrusion. In the presence of 2, 5-dimethyl-p-benzoquinone, net proton extrusion by Synechocystis sp. strain PCC6803 ceased after 1 min of illumination and a postillumination influx of protons was observed, suggesting that the PxcA-dependent, light-dependent proton extrusion equilibrates with a light-independent influx of protons. A photosystem I (PS I) deletion mutant extruded a large number of protons in the light. Thus, PS II-dependent electron transfer and proton translocation are major factors in light-driven proton extrusion, presumably mediated by ATP synthesis. Inhibition of CO2 fixation by glyceraldehyde in a cytochrome c oxidase (COX) deletion mutant strongly inhibited the proton extrusion. Leakage of PS II-generated electrons to oxygen via COX appears to be required for proton extrusion when CO2 fixation is inhibited. At pH 8.0, NO3- uptake activity was very low in the pxcA mutant at low [Na+] (approximately 100 microM). At pH 6.5, the pxcA strain did not take up CO2 or NO3- at low [Na+] and showed very low CO2 uptake activity even at 15 mM Na+. A possible role of PxcA-dependent proton exchange in charge and pH homeostasis during uptake of CO2, HCO3-, and NO3- is discussed.