WP2深海细菌基因组文库的构建和克隆子测序比对分析

以pUC19质粒DNA作为载体,用BamH1酶切、碱性磷酸酶处理后,分别与Sau3AⅠ部分酶切的Shewanella sp.(WP2) 基因组DNA 2-6kb的DNA片段连接,电击转化大肠杆菌DH5 α感受态,在含有Amp的LB平板上筛选重组子,得到转化子约5～6×103;随机挑取100个克隆子,经DNA测序,通过在网络数据库中进行BLASTX分析,得到一系列相对保守蛋白的比对结果.这些结果为遗传背景不清楚的细菌的研究,提供了一条从分子水平上探讨的技术路线.     

Shewanella algae对纯锌腐蚀性能的影响

【目的】在室内模拟条件下研究海藻希瓦氏菌(Shewanella algae)对纯锌牺牲阳极腐蚀的影响。【方法】利用微生物分析、交流阻抗测试技术、扫描电镜及荧光显微等方法,测试Shewanella algae的生长曲线、开路电位、电化学阻抗、表面形貌等。【结果】在含细菌体系中,细菌通过代谢活动消耗掉溶解氧,在试样表面形成一层生物膜。该生物膜阻挡了腐蚀介质与试样表面的接触,从而使试样的腐蚀受到抑制。荧光显微镜观察结果表明,浸泡7d,试样表面形成完整的生物膜,但随着营养物质及氧气的消耗,生物膜逐渐脱落。【结论】该细菌可以使纯锌试样的腐蚀电位升高,并且显著抑制试样的腐蚀。该细菌生物膜的形成需要一定的时间及充足的营养物质和氧气。     

Interactively interfacial reaction of iron-reducing bacterium and goethite for reductive dechlorination of chlorinated organic compounds

The interactively interfacial reactions between the iron-reducing bacterium （Shewanella decolorationis, S12） and iron oxide （α-FeOOH） were investigated to determine reductive dechlorination transformation of chlorinated organic compounds （chloroform and pentachlorophenol）. The results showed that the interactive system of S12＋ α-FeOOH exhibited relatively high dechlorination rate. By comparison, the S12 biotic system alone had no obvious dechlorination, and the α-FeOOH abiotic system showed low dechlorination rate. The enhanced dechlorination of chloroform and pentachlorophenol in the interactive system of S12＋α-FeOOH was derived from the promoted generation of adsorbed Fe（Ⅱ） by S12. A decrease in redox potential of the Fe （Ⅲ）/Fe （Ⅱ） couple in the interactive reaction system was determined by cyclic voltammetry. Our results will give new insight into interactively interfacial reaction between iron-reducing bacterium and iron oxides for degradation of chlorinated organic compounds under anaerobic condition.     

Shewanella sp.RQs-106还原Fe(Ⅱ)EDTA-NO产氨过程影响因素

考察了菌株Shewanella sp. RQs-106还原Fe(Ⅱ)EDTA-NO产氨过程的影响因素,在此基础上确定了适宜操作条件. 结果表明,乙酸钠和柠檬酸钠不能作为菌株RQs-106的电子供体,较适宜的电子供体为乳酸钠,在36 h内产氨率可达到99%. C/N质量比值过高或过低都会对菌株的Fe(Ⅱ)EDTA-NO降解率和产氨率产生影响,最适宜的C/N质量比值为20. Fe(Ⅱ)EDTA-NO浓度过高会抑制菌株活性,导致Fe(Ⅱ)EDTA-NO难以被降解,本实验条件下宜控制Fe(Ⅱ)EDTA-NO浓度为1 mmol/L. 当初始pH为5.0~6.0时,菌株RQs-106不能还原Fe(Ⅱ)EDTA-NO,当初始pH值升高到7.2~9.0时,可将Fe(Ⅱ)EDTA-NO还原为氨氮,但碱性过大对菌株RQs-106活性有抑制,较适宜的pH值为8.0. 菌株可在20~35 ℃内还原Fe(Ⅱ)EDTA-NO并产氨,最适温度为30 ℃.     

Screening and optimization of EPA-producing antarctic psychrophilic bacterium Shewanella sp. NJ136

Two hundred strains of bacteria from Antarctic sea ice were collected and screened for their ability of producing eieosapentaenoic acid （EPA, 20： 5ω3） by means of Gas Chromatography/Mass Spetmmetry （GC/MS）. Eight strains of bacteria containing EPA were investigated, among which the outstanding one was recorded as NJ136. This bacterium was identified as Shewanella by the biochemical characteristics and 16S rRNA sequence. Response surface methodology （RSM） was applied to optimize the medium ingredients. A 24 full factorial central composite design （FFCCD） was employed to determine the naximum EPA production at optimum levels of pH, NaC1, glucose and yeast extract. The predicted optimal combination of media constituents for maximum 14.02 mg/g （ about 1.7-fold increase） EPA production were determined as 30.15%e （m/v） NaC1, 9.98 g/L glucose, 4.42 g/L yeast extract and pH 6.08. The actual experimental results were in agreement with the prediction.     

氧化还原介体催化强化Shewanellaoneidensis MR-1还原亚硒酸盐的研究

针对微生物还原亚硒酸盐过程普遍存在的时间较长的问题,考察了4种水溶性醌类介体(α-AQS,AQS,1,5-AQDS和AQDS)对奥奈达希瓦氏菌(Shewanella oneidensis MR-1)还原亚硒酸盐过程的加速作用,通过单因素试验优化了培养条件,对硒纳米颗粒的Zeta电位和粒径进行了表征。结果表明,4种醌类介体都加速了亚硒酸盐的还原,其中,AQDS的加速效果最显著;在pH值为8.0,温度为30℃,AQDS浓度为0.2mmol/L条件下,48h时亚硒酸盐的转化率达到100%;加入AQDS后会生成更大尺寸的硒纳米颗粒,并可能使硒纳米颗粒表面包裹的有机物质成分及含量发生改变。研究结果为Shewanella oneidensis MR-1修复亚硒酸盐污染的实际应用提供了理论依据。     

Shewanella haliotis BP-1海藻酸裂解酶基因的克隆表达

【目的】了解海洋细菌Shewanella haliotis BP-1中海藻酸裂解酶降解海藻酸钠的生物活性。【方法】应用基因克隆和大肠杆菌异源表达技术,过量表达海藻酸裂解酶,将粗酶液通过DEAE Sepharose FF柱分离纯化后检测其酶活性。【结果】从S.haliotis BP-1菌株的基因组DNA中克隆得到一个大小为2 157bp的海藻酸裂解酶基因Alg17S,该基因编码的海藻酸裂解酶Alg17S属于PL17家族的蛋白,大小为79 726Da,其中包括N端26个氨基酸的信号肽,与Saccharophagus degradans 2-40菌株产生的海藻酸裂解酶Alg17C具有高度同源性,相似性为52%。经纯化后获得的重组酶Alg17S和△snAlg17S(N端不含26个氨基酸的信号肽)均具有降解海藻酸钠的活性,但△snAlg17S对海藻酸钠的催化活性比Alg17S高,其酶比活力高达9 635U/mg。【结论】重组海藻酸裂解酶△snAlg17S兼具高表达水平及高酶活性,是进一步研究海藻酸盐糖化和生物燃料生产的潜在的优势酶。