耐铅镉菌株的分离鉴定及其吸附能力

堆肥中添加生物钝化剂是当前降低粪便中重金属生物毒性最为有效的方式之一,为了进一步提高其钝化重金属的能力,文中获得了复合重金属高耐性的钝化剂菌株,并探究其生物学特性和吸附特征。采集猪粪堆肥样品并在改良的牛肉膏培养基中分离和筛选耐铅又耐镉的高耐性菌株,通过形态结合分子生物学鉴定该菌株。该菌株分别在不同pH、温度和盐浓度条件下培养获得其最适的生长条件,进而在该条件下分析其对铅镉吸附的特性。结果获得一株耐铅浓度为600 mg/L、镉浓度为120 mg/L的铅镉复合耐性菌株SC19,该菌株为西地西菌属,其最适生长环境为pH值7.0、温度37℃、盐浓度0.5%。培养36 h的稳定期SC19菌株在吸附时间30min时铅的去除率最高,对铅的最大去除率和吸附量分别为60.7%和329.13mg/g;培养8h的对数期在吸附时间30min时镉的去除率最高,对镉的最大去除率和吸附量分别为51.0%和126.19 mg/g。红外光谱分析显示,SC19菌株对铅镉离子的吸附主要是细胞表面的多种活性基团与金属离子发生络合作用。该研究揭示了SC19菌株有较好的二价态铅镉离子吸附能力,可为生物钝化重金属提供重要的微生物种质资源。     

整肠生菌纯培养物的环境动力学研究

采用振荡培养研究了温度、PH值和不同培养基的环境因素对整肠生菌（地衣芽孢杆菌）消长的影响．通过对整肠生菌的动力学基础数据按计算作图的动力学研究结果表明：整肠生菌最适生长温度在35～37℃时，活菌数15─17.2×10~7／ml,是30℃和40℃时的1.26倍和1.2倍．最适PH值在5.5～7.5时,活菌数为4.15─7.55×10~8／ml,是PH值4.5和9.5时的13.l倍和13.2倍．ABC三种培养基的结果表明,培养基C最适合地衣芽孢杆菌生长，菌数最高，为69.05×10~8／ml．对数期的世代时间也最短，G=20min,比AB号培养基中培养时缩短2─6min．生长比速度（μ=0.035)最大，是AB培养基培养时的1.27倍．残废率较低,5.2％．环境因素与整肠生菌消长的这种动力学关系，为利用地衣芽孢杆菌发酵生产“整肠生”制剂的调控以及该菌剂的治疗机制方面研究提供了科学依据．     

一株刚果红降解菌的筛选、鉴定和性质的初步研究

通过稀释划线的方法,从土壤中分离到一株能降解刚果红的菌株(T1),从菌落和孢子形态来判断该菌为放线菌的链霉菌属。T1菌在含刚果红(100 mg.L-1)高氏液体培养基中培养6 d后,脱色率高达90%。对培养液进行200~800 nm波长扫描的结果表明,培养基中部分刚果红被T1菌降解,其余部分被菌体吸附。活菌体对染料的吸附效率比死菌体高。T1菌对刚果红的脱色主要是通过生物降解和菌体吸附作用来完成。     

不同钾浓度培养条件对栖热茵TibetanG6菌株吸附铯的影响

研究了不同钾离子浓度培养体系下栖热菌TibetanG6菌株(Thermus sp．TibetanG6)对铯的吸附规律以及pH和钠离子对吸附的影响．结果表明,不同钾离子浓度培养条件在TibetanG6菌株对铯的吸附过程中扮演了重要的角色,由培养基中不加钾而诱导的钾缺失细胞对铯的吸附量(24 h)明显高于正常培养基中加过量钾产生的钾过量体系中的吸附量．在不同的钾离子细胞中,pH和钠离子对菌体吸附铯产生了不同的影响．菌体的红外光谱分析也表明：菌株TibetanG6细胞对铯的吸附是一个动态平衡过程．最后对pH和钠离子对铯吸附的作用机制进行了讨论．     

出芽短梗霉菌JB16铅镉吸附特性的研究北大核心

【目的】本文对污染土壤中的耐重金属菌株进行分离鉴定,研究菌株在不同条件下对吸附铅镉的影响因素。【方法】通过生理生化特征及ITS序列分析确定菌株种属,采用平板划线法确定最大耐铅镉浓度并探究菌株吸附的最佳条件;通过准二级动力学、Langmuir和Freundlich等温吸附的模型及红外光谱探究吸附过程。【结果】菌落形态和ITS序列分析鉴定表明,筛选分离的JB16为出芽短梗霉菌(Aureobasidium pullulans),最大耐铅浓度达1500 mg/L,最大耐镉浓度达750 mg/L,最大耐铅镉混合浓度达1500 mg/L和300 mg/L。通过单因素实验(温度、时间、菌龄、pH、湿菌体浓度和初始重金属浓度)得出结论,在温度30℃、时间2 h、菌龄72 h、pH 6、湿菌体浓度5 g/L和初始铅浓度150 mg/L的最佳条件下,菌体对铅的吸附率为88.5%;在温度30℃、时间1 h、菌龄96 h、pH 6、湿菌体浓度5 g/L和初始镉浓度20 mg/L的最佳条件下,菌体对镉的吸附率为59.4%。菌株吸附铅镉过程符合Langmuir吸附模型和准二级动力学模型,为表面单分子层吸附。扫描电镜和红外光谱分析表明,重金属离子对菌体造成影响,吸附前后形态发生变化,细胞表面的羟基、羧基、饱和C−H键和酰胺基等基团参与了吸附过程。【结论】菌株JB16具有一定的铅镉吸附效果,为修复重金属铅镉污染的水体和土壤提供宝贵的菌种资源和数据支持。     

植酸钠对酿酒酵母乙醇发酵的促进效应

本文研究了植酸钠对酿酒酵母发酵乙醇生成量的促进效应。在酿酒酵母（ＡＳ２．４３１）的合成培养基中添加０．０８％本室自制的植酸钠后,发现其乙醇生成量提高３５．４％。植酸钠使该菌耐酒精能力增加,但对生物量影响不显著。植酸钠促进效应的有效成分为其植酸根。植酸钠对Ｋ酵母天然培养基发酵的乙醇生成量亦有极显著的促进效应,使其提高达８７．９％。     

一株耐铅镉真菌的分离鉴定及其吸附特性的研究

【目的】以扎龙湿地污染的土壤为材料,进行耐铅镉菌株的分离鉴定,研究不同条件对菌株吸附铅镉的影响。【方法】采用平板划线法,逐级驯化,筛选出一株耐铅镉菌株,通过生理生化特征及ITS序列分析对菌株进行鉴定,探究该菌吸附的最佳条件,并进行Langmuir和Freundlich等温吸附模型拟合。【结果】本研究分离得到一株菌株JB15,最高耐受浓度为Pb²⁺1200 mg/L、Cd²⁺200 mg/L,经鉴定为球孢白僵菌,最佳吸附条件温度为30°C,pH为7.0,接菌量为8.0 g/L,吸附时间为60 min,铅镉吸附率分别为52.27%和62.38%;铅镉吸附量分别为19.60 mg/g和3.98 mg/g,符合Langmuir等温吸附模型。【结论】菌株JB15具有较好的吸附效果,可为微生物修复重金属土壤污染提供理论基础。     

海洋破囊壶菌Δ4-脂肪酸脱饱和酶基因在酿酒酵母中的表达

以质粒pGEM-TFAD4为模板,扩增获得1．6kb的△^4-脂肪酸脱饱和酶基因（FAD4）。将FAD4酶切后连接到hin dⅢ,Xba I处理过的pYEs2．0载体,构建重组表达质粒pYFAD4。转化酿酒酵母缺陷型菌INVScl,通过SC-U选择性培养基筛选阳性克隆子。添加外源脂肪酸C22：5底物,半乳糖诱导表达。气相色谱分析表明阳性克隆子总脂肪酸中出现了二十二碳六烯酸C22：6（占酵母总脂肪含量的41．13％）,△^4-脂肪酸脱饱和酶基因在酿酒酵母中得到了表达。     

非生长产黄青霉吸附铅的研究

本文报道非生长产黄青霉(Penicillium chrysogenum)对溶液中铅离子的吸附。菌体对铅的吸附依赣于溶液的pH值,在pH4-5范围内,其饱和吸附量(116mgPb2+／g干菌)高于活性炭及文献中所报道的蕈状芽抱仟菌(Bacillus mycoidrs),小刺青(Penicillium spinulosum)及长木链霉(Streptomyces longwoodensis)等。当pH4．5时,产黄青霉对铅的选择性高于Zn2+、Cd2+,Cu2+和As3+Cu2+、As3+的存在对铅的吸附无明显影响。Cd2+的存在使铅的吸附量有所提高．而Zn2+的存在则使之降低。在小试规模下,经本法处理过的含铅工业废水,铅含量降至1.0mg／1．以下,达到国家规定的工业废水铅含量的排放标准。     

球衣菌对工业废水中Cu2+吸附条件的研究

利用球衣菌FQ32制备生物吸附剂,研究其对Cu^2＋的耐受能力,并以连江某工厂含Cu^2＋废水为样液,对其进行吸附条件试验,分析了影响吸附率和吸附量的因素,结果表明,球衣菌FQ32对Cu^2＋的耐受能力大于150mg／L。在工业废水中Cu^2＋的浓度为23．1mg／L,吸附剂用量为0．2g／L,30℃,pH值6．0,吸附时间10min的条件下,该吸附剂对Cu^2＋的吸附率达到85．2％,吸附量达91．21mg／g。     

Biosorption of heavy metals by Saccharomyces cerevisiae: a review

Heavy metal pollution has become one of the most serious environmental problems today. Biosorption, using biomaterials such as bacteria, fungi, yeast and algae, is regarded as a cost-effective biotechnology for the treatment of high volume and low concentration complex wastewaters containing heavy metal(s) in the order of 1 to 100 mg/L. Among the promising biosorbents for heavy metal removal which have been researched during the past decades, Saccharomyces cerevisiae has received increasing attention due to the unique nature in spite of its mediocre capacity for metal uptake compared with other fungi. S. cerevisiae is widely used in food and beverage production, is easily cultivated using cheap media, is also a by-product in large quantity as a waste of the fermentation industry, and is easily manipulated at molecular level. The state of the art in the field of biosorption of heavy metals by S. cerevisiae not only in China, but also worldwide, is reviewed in this paper, based on a substantial number of relevant references published recently on the background of biosorption achievements and development. Characteristics of S. cerevisiae in heavy metal biosorption are extensively discussed. The yeast can be studied in various forms for different purposes. Metal-binding capacity for various heavy metals by S. cerevisiae under different conditions is compared. Lead and uranium, for instances, could be removed from dilute solutions more effectively in comparison with other metals. The yeast biosorption largely depends on parameters such as pH, the ratio of the initial metal ion and initial biomass concentration, culture conditions, presence of various ligands and competitive metal ions in solution and to a limited extent on temperature. An assessment of the isotherm equilibrium model, as well as kinetics was performed. The mechanisms of biosorption are understood only to a limited extent. Elucidation of the mechanism of metal uptake is a real challenge in the field of biosorption. Various mechanism assumptions of metal uptake by S. cerevisiae are summarized.     

Biosorption with algae: a statistical review

The state of the art in the field of biosorption using algae as biomass is reviewed. The available data of maximum sorption uptake (qmax) and biomass-metal affinity (b) for Cd2 +, Cu2 +, Ni2 +, Pb2 + and Zn2 + were statistically analyzed using 37 different algae (20 brown algae, 9 red algae and 8 green algae). Metal biosorption research with algae has used mainly brown algae in pursuit of treatments, which improve its sorption uptake. The information available in connection with multimetallic systems is very poor. Values of qmax were close to 1 mmol/g for copper and lead and smaller for the other metals. Metal recovery performance was worse for nickel and zinc, but the number of samples for zinc was very small. All the metals except lead present a similar affinity for brown algae. The difference in the behavior of lead may be due to a different uptake mechanism. Brown algae stand out as very good biosorbents of heavy metals. The best performer for metal biosorption is lead.     

Functional expression of the voltage-gated neuronal mammalian potassium channel rat ether à go-go1 in yeast.

It has been shown previously that heterologous expression of inwardly rectifying potassium channels (K+-channels) from plants and mammals in K+-transport defective yeast mutants can restore the ability of growth in media with low [K+]. In this study, the functional expression of an outward rectifying mammalian K+-channel in yeast is presented for the first time. The outward-rectifying mammalian neuronal K+-channel rat ether à go-go channel 1 (rEAG1, Kv 10.1) was expressed in yeast (Saccharomyces cerevisiae) strains lacking the endogenous K+-uptake systems and/or alkali-metal-cation efflux systems. It was found that a truncated channel version, lacking almost the complete intracellular N-terminus (rEAG1 Delta 190) but not the full-length rEAG1, partially complemented the growth defect of K+-uptake mutant cells (trk1,2 Delta tok1 Delta) in media containing low K+ concentrations. The expression of rEAG1 Delta 190 in a strain lacking the cation efflux systems (nha1 Delta ena1-4 Delta) increased the sensitivity to high monovalent cation concentrations. Both phenotypes were observed, when rEAG1 Delta 190 was expressed in a trk1,2 Delta and nha1, ena1-4 Delta mutant strain. In the presence of K+-channel blockers (Cs+, Ba2+ and quinidine), the growth advantage of rEAG1 Delta 190 expressing trk1,2 tok1 Delta cells disappeared, indicating its dependence on functional rEAG1 channels. The results demonstrate that S. cerevisiae is a suitable expression system even for voltage-gated outward-rectifying mammalian K+-channels.     

Characterization of acetone-washed yeast biomass functional groups involved in lead biosorption

The mechanism of lead cation biosorption by acetone-washed biomass of Saccharomyces uvarum was investigated by chemical modifications and spectroscopic monitoring of the cell components. Reacting the carboxyl groups with propylamine, which neutralizes these anions, considerably decreased the metallic ion uptake, indicating that negatively charged carboxyl groups play an important role in lead bisorption due to electrostatic attraction. After lead biosorption the photoacoustic Fourier transform infrared spectroscopy showed a change in the symmetrical stretch of the carboxylate groups of the acetone-washed yeast biomass, and the X-ray photoelectron spectroscopy oxygen peak was also found to be shifted. These findings support the hypothesis that lead uptake occurs mainly through binding to the carboxyl group. In X-ray photoelectron spectroscopy the nitrogen peak decreased after the biosorption of lead, suggesting that nitrogen-containing groups are also involved in the biosorption process. Acylation of amino groups was shown to increase the lead biosorption capacity. The acylation reaction converts the positively charged amino group to an amide capable of coordination to lead cations. Deproteination by boiling the biosorbent with NaOH increased the lead uptake. The acetone-washed biomass uptake of lead from an aqueous solution at ph 5.5 was 48.9 mg/g dry weight. Pure chitin adsorbed 48.8 mg lead/g dry weight. Mannan isolated from S. uvarum did not adsorb lead at all. Electrostatic attraction of the carboxyl groups and other anions present in the acetone-washed biomass, and complexation with nitrogen atoms, especially in chitin, appear to be the main mechanisms involved in lead cation biosorption. (c) 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 55: 1-10, 1997.     

Biosorption with Algae: A Statistical Review

ABSTRACT

The state of the art in the field of biosorption using algae as biomass is reviewed. The available data of maximum sorption uptake (q_max) and biomass-metal affinity (b) for Cd^{2 +}, Cu^{2 +}, Ni^{2 +}, Pb^{2 +} and Zn^{2 +} were statistically analyzed using 37 different algae (20 brown algae, 9 red algae and 8 green algae). Metal biosorption research with algae has used mainly brown algae in pursuit of treatments, which improve its sorption uptake. The information available in connection with multimetallic systems is very poor. Values of q_max were close to 1 mmol/g for copper and lead and smaller for the other metals. Metal recovery performance was worse for nickel and zinc, but the number of samples for zinc was very small. All the metals except lead present a similar affinity for brown algae. The difference in the behavior of lead may be due to a different uptake mechanism. Brown algae stand out as very good biosorbents of heavy metals. The best performer for metal biosorption is lead.     

Changes in 45Ca and 109Cd uptake, membrane potential and cell pH in Saccharomyces cerevisiae provoked by Cd2+

The effect of Cd2+ poisoning of Saccharomyces cerevisiae on 45Ca, 109Cd and [14C]tetraphenylphosphonium (TPP) uptake and cell pH was examined. At Cd2+ concentrations that produced substantial K+ efflux the rates of uptake of 45Ca, 109Cd and [14C]TPP increased progressively during incubation of the cells with Cd2+, and the cell pH was lowered concomitantly. The initial rates of uptake of the divalent cations and of TPP were increased in cells pre-loaded with Cd2+, which shows that stimulation of the ion fluxes was exerted by the Cd2+ that accumulated in the cells. The distribution ratio of TPP between cells and medium, however, was decreased by Cd2+. Although hyperpolarization of the cell membrane by Cd2+ cannot be excluded, it is argued that Cd2+ primarily stimulated divalent cation uptake by increasing the cation permeability of the cell membrane allowing the cations to enter the cells more easily.     

Static magnetic fields enhancement of Saccharomyces cerevisae ethanolic fermentation

Magnetic effects induced in ethanolic fermentation by Saccharomyces cerevisiae strain DAUFPE-1012 were studied during a 24 h exposure to 220 mT steady magnetic fields (SMF) at 23 +/- 1 degrees C, produced by NdFeB rod magnets. The magnets were attached diametrically opposed (N to S) to a cylindrical tube reactor. The biomass growth in the reactor culture media (yeast extract + glucose 2%) during 24 h was monitored by measurements of optical density, which was correlated to cell dry weight. Ethanol concentration and glucose level were measured every 2 h. The pH of the culture media was maintained between 4 and 5. As a result, biomass (g/L) increased 2.5-fold and ethanol concentration 3.4-fold in magnetized cultures (n = 8) as compared with SMF nonexposed cultures (n = 8). Glucose consumption was higher in magnetized cultures, which correlated to the ethanol yield.     

转HAL1基因番茄的耐盐性

利用农杆菌介导的叶盘法,把HAL1 基因转入番茄,Southern杂交检测得到转基因植株.耐盐实验表明, T1代转基因番茄在150 mmol/L的NaCl胁迫下仍有43%的发芽率,200 mmol/L的NaCl胁迫下发芽率为6%,而对照种子在100和150 mmol/L的NaCl胁迫下发芽率分别为11.0%和0.转基因番茄的电解质相对外渗率小于对照,而根冠比和叶绿素含量大于对照,转HAL1基因显著提高了番茄的耐盐性.盐胁迫下Na 、K 的累积状况表明,转基因番茄根、茎、叶的K /Na 均有所提高,根系的SK/Na增大,茎、叶的RSK/Na和RLK/Na减小,说明根系对K /Na 离子的选择吸收和运输能力加强.不但选择吸收K /Na ,而且表现出整株水平上的有利于耐盐的K /Na 区域化分配.     

The Influence of Calcium or Manganese on the Resistance to EDTA, Polymyxin B or Cold Shock, and the Composition of Pseudomonas aeruginosa Grown in Glucose- or Magnesium-depleted Batch Cultures

Cultures were batch grown in simple salts media in which growth was limited either by depletion of glucose and magnesium (C/Mg-dep) or by glucose alone (C-dep). Cultures were also grown in these media supplemented by calcium and/or manganese.
All cultures grown in the C-dep media were sensitive to ethylenediaminetetraacetic acid (EDTA), polymyxin and also to cold shock but were relatively resistant to ethyleneglycol-bis(2-aminoethyl ether)-N, N-tetraacetic acid (EGTA). Inclusion of calcium or manganese in the growth medium enhanced lysis by EDTA. Cultures grown in the basic C/Mg-dep media were resistant to EDTA, EGTA, polymyxin and to cold shock. Sensitivity to these agents was retained by cultures grown in C/Mg-dep media supplemented with Ca²⁺ and/or Mn²⁺. Cells grown in C/Mg-dep media with added Mn²⁺ were more sensitive to EDTA and polymyxin than those from the unsupplemented C/Mg-dep media but still resistant compared with C-dep cultures. All cultures from supplemented C/Mg-dep media were more sensitive to EGTA than those from any of the C-dep media.
Whole cells and cell walls from these various media had differing amounts of cell wall, phosphorus, amino sugar, carbohydrates, readily extractable lipid (REL), total phospholipid (PL), and especially differences in cell wall divalent metal cation content.
The differences in PL, REL and amino sugars and carbohydrate did not correlate with the response of C-dep and C/Mg-dep bacteria to EDTA, EGTA or polymyxin. The results are discussed in relation to the hypothesis that the sensitivity of Pseudomonas aeruginosa to polymyxin and EDTA is more dependent on outer membrane cation content rather than on other components, e.g. PL and lipopolysaccharide.