Intracellular recovery of gold by microbial reduction of AuCl4 ions using the anaerobic bacterium Shewanella algae

Microbial reduction and intracellular precipitation of gold was achieved at 25 °C and pH 7 by using the mesophilic anaerobic bacterium Shewanella algae with H₂ as the electron donor. The reductive precipitation of gold by S. algae was a fast process: 0.1–1 mol/m³ AuCl₄⁻ ions were completely reduced to insoluble gold within 30 min. The biogenic precipitates were crystalline gold nanoparticles of 10–20 nm present in the periplasmic space. The reducing power of S. algae at 3.2 × 10¹⁵ cells/m³ and 25 °C was comparable to that of aqueous citric acid solution (chemical reductant) at 20 mol/m³ and 50 °C. The intracellular recovery of gold is potentially attractive as an environmentally friendly alternative to conventional methods.     

Spoilage of refrigerated (4 °C) Litopenaeus vannamei: cooperation between Shewanella species and contribution of cyclo‐(L‐Pro‐L‐Leu)‐dependent quorum sensing

The spoilage potential of Shewanella putrefaciens and S. baltica isolated from spoiled refrigerated Litopenaeus vannamei was evaluated by in vitro assays for trimethylamine oxide reduction, extracellular hydrolytic enzymes and biofilm formation, and in vivo inoculation into surface‐sterilised shrimp followed by microbial, biochemical and sensory analyses during storage for 5 days at 4 °C. S. baltica displayed higher spoilage potential than S. putrefaciens both in vitro and in vivo. Shrimp co‐inoculated with them had one‐day shorter shelf life than those mono‐inoculated, based on the results of bacterial density, volatile base nitrogen, trimethylamine, volatile organic compounds and sensory analysis, which strongly suggests cooperation of Shewanella species in shrimp spoilage. Exogenous cyclo‐(L‐Pro‐L‐Leu) boosted bacterial growth, extracellular protease and collagenase activities, and biofilm formation of S. putrefaciens and S. baltica at least before they entered the stationary phase, indicating that cyclo‐(L‐Pro‐L‐Leu)‐dependent quorum sensing, a recently suggested communication mechanism between them, contributes to the cooperation.     

Three Microbial Musketeers of the Seas: Shewanella baltica,Aliivibrio fischeri and Vibrio harveyi,and Their Adaptation to Different Salinity Probed by a Proteomic Approach

Osmotic changes are common challenges for marine microorganisms. Bacteria have developed numerous ways of dealing with this stress, including reprogramming of global cellular processes. However, specific molecular adaptation mechanisms to osmotic stress have mainly been investigated in terrestrial model bacteria. In this work, we aimed to elucidate the basis of adjustment to prolonged salinity challenges at the proteome level in marine bacteria. The objects of our studies were three representatives of bacteria inhabiting various marine environments, Shewanella baltica, Vibrio harveyi and Aliivibrio fischeri. The proteomic studies were performed with bacteria cultivated in increased and decreased salinity, followed by proteolytic digestion of samples which were then subjected to liquid chromatography with tandem mass spectrometry analysis. We show that bacteria adjust at all levels of their biological processes, from DNA topology through gene expression regulation and proteasome assembly, to transport and cellular metabolism. The finding that many similar adaptation strategies were observed for both low- and high-salinity conditions is particularly striking. The results show that adaptation to salinity challenge involves the accumulation of DNA-binding proteins and increased polyamine uptake. We hypothesize that their function is to coat and protect the nucleoid to counteract adverse changes in DNA topology due to ionic shifts.     

凡纳滨对虾优势腐败菌鉴定及其致腐能力的初步研究

分析鉴定了凡纳滨对虾0℃与20℃贮藏条件下的菌相组成与优势腐败菌,并对优势腐败菌16SrDNA、生长动力学、致腐能力与菌落数的变化进行了测定。结果表明,0℃与20℃贮藏条件下,对虾优势腐败菌分别是希瓦氏菌(30%)、不动杆菌(16.7%)与希瓦氏菌(46.5%)、发光杆菌(17.7%)。7℃条件下,将一定浓度的希瓦氏菌与不动杆菌菌悬液接种到无菌对虾上,结果显示接种希瓦氏菌的样品其腐败代谢产物产量因子YTVB-N/CFU、YTMA/CFU分别为12.44×10-9、6.193×10-10,而接种不动杆菌的样品其YTVB-N/CFU、YTMA/CFU分别为8.937×10-9、5.548×10-10。结果表明,7℃条件下,希瓦氏菌的致腐能力强于不动杆菌,希瓦氏菌在对虾腐败过程中占主导作用,其分析结果与对虾菌相组成的鉴定结果相一致。     

Optimization of alkaline protease production from Shewanella oneidensis MR‐1 by response surface methodology

BACKGROUND: The proteases are among the most important groups of enzymes. Therefore, it is important to produce inexpensive and optimized media for large‐scale commercial production. In the present work, three different Shewanella species were screened on skim milk agar medium for their ability to produce alkaline protease. The effects of different culture conditions were optimized for alkaline protease production by S. oneidensis MR‐1 using a Box–Behnken design combined with response surface methodology (RSM). RESULTS: Highest yield (112.90 U mL^?1) of protease production was obtained at pH 9.0, a temperature of 30 °C, glucose (12.5 g L^?1), tryptone (12.5 g L^?1) and an incubation period of 36 h. A second‐order polynomial regression model was used for analysis of the experiment. The experimental values were in good agreement with predicted values, with correlation coefficient 0.9996. CONCLUSION: Carbon and nitrogen, pH, temperature and incubation period were chosen as the main factors to be used in an experimental design for optimization to produce low‐cost enzymes, potentially for use on an industrial scale. A 60% increase in enzyme activity was achieved in the optimized medium compared with the original medium. Copyright © 2008 Society of Chemical Industry     

不同细菌胞外产物对腐败希瓦氏菌低温下生长代谢的影响

为研究荧光假单胞菌(Pseudomonas fluorescens,P.fluorescens)、嗜水气单胞菌(Aeromonas hydrophila,A.hydrophila)、温和气单胞菌(Aeromonas sobria,A.sobria)的胞外产物对腐败希瓦氏菌(Shewanella putrefaciens,S.putrefaciens)低温下生长的影响,提取这3株菌株的胞外产物与无菌脑心浸肉汤(Brain Heart Infusion,BHI)培养基混合,再将S.putrefaciens分别接种至混合培养液中,于(4±1)℃下培养12d,测定培养液中S.putrefaciens的菌落总数及胞外蛋白酶、生物膜、挥发性盐基氮(TVB-N)、总氨基酸和腐胺的变化。结果表明:P.fluorescens、A.hydrophila和A.sobria的胞外产物对S.putrefaciens的延滞期和对数期几乎没有影响,但是A.sobria的胞外产物可以抑制S.putrefaciens稳定期的生长,而且随着贮藏时间的延长,相比于P.fluorescens、A.hydrophila,A.sobria的胞外产物对S.putrefaciens的生物膜、胞外蛋白酶形成以及致腐败能力的抑制作用更强。     

大黄鱼来源波罗的海希瓦氏菌SB-19的hfq基因功能分析

为分析大黄鱼来源波罗的海希瓦氏菌（Shewanella baltica）中分子伴侣Hfq的功能,对具有强致腐能力的波罗的海希瓦氏菌SB-19中hfq基因进行敲除,比较野生株（wide type,WT）和敲除株（knock out,KO）在生长速率、群体感应、抗逆境以及对灭菌鱼汁致腐效果的差异。结果表明：波罗的海希瓦氏菌SB-19中存在1 个hfq基因,其基因表达水平随着细胞密度的增加而升高;在30 ℃培养条件下,WT株和KO株的生长速率接近,但在4 ℃培养条件下,与WT株相比,KO株的生长出现明显迟滞且稳定期的细胞密度较低;WT株与KO株分泌信号分子的能力相近,但KO株表现出较低的生物被膜形成能力和胞外蛋白酶活性,而且接种KO株的灭菌鱼汁也产生较少的挥发性盐基氮和三甲胺;此外,KO株对盐分、营养、重金属、消毒剂等胁迫条件更为敏感。上述结果表明Hfq参与波罗的海希瓦氏菌SB-19株的多种代谢调控,影响细菌的生长、群体感应、致腐能力、抗逆境能力等,即Hfq是一种全局性调控因子,整体调节波罗的海希瓦氏菌SB-19的诸多生理活动。本实验为波罗的海希瓦氏菌的基础生物学研究和海产品中微生物的致腐机制提供理论基础。     

胞外聚合物在腐败希瓦氏菌处理铀污染地下水中的作用

以模拟铀污染地下水为对象,采用腐败希瓦氏菌对其进行了固定铀实验,利用单粒子 电感耦合等离子体质谱（SP ICP MS）、3种胞外聚合物（EPS）提取方法和碳酸氢钠 硝酸分级提取法表征了腐败希瓦氏菌和EPS中铀的含量和赋存形态,并采用傅立叶红外光谱（FT IR）技术分析了EPS与铀反应前后的变化。结果表明,阳离子交换树脂（CER）法适用于表征EPS中的铀含量及形态,EPS中的铀可占腐败希瓦氏菌固定铀总量的(140±10)%。SP ICP MS分析表明,CER法获得的EPS中颗粒铀是其主要形态,EPS在固定铀的过程中,既涉及到生物吸附,也涉及到生物矿化。FT IR分析进一步说明生物吸附时,铀可与EPS中的羧基官能团结合。以上结果表明,EPS在腐败希瓦氏菌固定铀的过程中发挥了重要作用。     

原位合成纳米SiOx/LZM/TP-CS涂膜对腐败希瓦氏菌的抑菌机理

目的 为拓展壳聚糖(CS)保鲜涂膜在水产品中的应用,并为水产品保鲜机制的研究提供理论支持.方法 以原位合成纳米SiOx改性的CS为成膜基质,以溶菌酶(LZM)和茶多酚(TP)为添加剂,采用流延法制备复合保鲜涂膜.以水产品中优势腐败菌——腐败希瓦氏菌为抑菌对象,研究涂膜的抑菌性能,用扫描电子显微镜(SEM)观测复合保鲜涂膜处理前后菌体的微观形貌变化,测定复合保鲜涂膜处理一定时间后菌悬液的电导率、OD260值、Na+/K+-ATP酶和AKP酶活性的变化.结果 各涂膜均表现出了抑菌性能,涂膜处理后菌体变形,且其菌悬液的电导率、OD260值、Na+/K+-ATP酶和AKP酶活性明显高于同期未处理样品.CS经纳米SiOx改性后,涂膜的抑菌性能略有提高,菌体多处塌陷,菌悬液电导率、OD260值、Na+/K+-ATP酶和AKP酶活性显著增加;继续加入LZM或TP后,涂膜的抑菌性能继续提高,且原位合成纳米SiOx/TP-CS涂膜抑菌性能更优.LZM和TP同时加入经纳米SiOx改性的CS涂膜中后,复合涂膜处理的菌体细胞壁溶解,损伤严重,同期处理的菌悬液电导率、OD260值最高,并对Na+/K+-ATP酶、AKP酶活性有抑制作用.结论 结合前期的研究结果,SiOx/LZM/TP-CS复合保鲜涂膜具有良好的保鲜、抑菌性能,可为可食性保鲜涂膜的应用提供理论指导和技术支持.     

培养温度对腐败希瓦氏菌DSM6067生长动力学及细胞膜理化特性的影响

低温是保障水产品质量安全的重要方法,然而仍有部分耐冷菌能适应低温环境,继续生长繁殖并最终导致水产品腐败变质。为了解不同培养温度对腐败希瓦氏菌生长动力学参数及细胞膜理化特性变化的影响,以水产品中典型的耐冷腐败微生物腐败希瓦氏菌模式菌株DSM6067为研究对象,对细菌的生长情况、细胞膜脂肪酸组成、膜蛋白含量、细胞微观结构和细胞膜流动性进行分析。结果表明,腐败希瓦氏菌模式菌株DSM6067分别在30,10℃和4℃培养条件下经3.35,25.94 h和122.03 h的延滞期后进入对数生长期;腐败希瓦氏菌虽在低温环境下培养增速较为缓慢,但能够适应低温环境继续生长。腐败希瓦氏菌在相应培养温度下的对数生长中期,30℃条件下细菌细胞膜不饱和脂肪酸总量仅43.36%,其中C16:1(棕榈油酸)为26.62%,而4℃培养下细菌细胞膜不饱和脂肪酸总量为51.04%,其中单不饱和脂肪酸C16:1含量高达46.66%,低温培养下细菌细胞膜中C16:1含量显著增加;细菌细胞膜蛋白表达量随培养温度的降低而有所上升;腐败希瓦氏菌微观结构观察显示,低温培养下的细菌细胞膜较为平滑,胞内空泡较少,细胞体积略大;在低温适应过程中,细菌细胞膜流动性增强。腐败希瓦氏菌细胞膜理化特性所产生的变化为细菌更好地适应低温环境生长提供了一定的物质基础。