This paper describes the investigation of the secretion of extracellular polymeric substances(EPS)by an extremely thermoacidophilic archaea,Metallosphaera sedula(M.sedula),during the bioleaching of pyrite under differ...This paper describes the investigation of the secretion of extracellular polymeric substances(EPS)by an extremely thermoacidophilic archaea,Metallosphaera sedula(M.sedula),during the bioleaching of pyrite under different temperatures and discusses the relationship among the EPS secretion,its heat resistance,and its ability to bioleach pyrite.The investigation results indicate that the amount of extracellular proteins is significantly higher than the amount of extracellular polysaccharides in the extracted EPS whether free cells or attached cells;these results are quite different from the behavior of mesophilic Acidithiobacillus ferrooxidans.Although the growth of M.sedula is inhibited at 80℃,the bioleaching ability of M.sedula is only slightly lower than that at the optimum growth temperature of 72℃ because of the heat resistance mechanism based on EPS secretion.The secretion of more extracellular proteins is an important heat resistance mechanism of M.sedula.展开更多
Lipid-producing microalgae are emerging as the leading platform for producing alternative biofuels in response to diminishing petroleum reserves. Optimization of fatty acid production is required for efficient convers...Lipid-producing microalgae are emerging as the leading platform for producing alternative biofuels in response to diminishing petroleum reserves. Optimization of fatty acid production is required for efficient conversion of microalgal fatty acids into usable transportation fuels. Microbial lipases/esterases can be used to enhance fatty acid production because of their efficacy in catalyzing hydrolysis of esters into alcohols and fatty acids while minimizing the potential poisoning of catalysts needed in the biofuel production process. Although studies have extensively focused on lipases/esterases produced by mesophilic organisms, an understanding of lipases/esterases produced by thermophilic, acidic tolerant microbes, such as Metallosphaera sedula, is limited. In this work, the carboxylesterase from Metallosphaera sedula DSM5348 encoded by Msed_1072 was recombinantly expressed in Escherichia coli strain BL21 (λDE3). The purified enzyme either with a hexahistidine (His6)-tag (Msed_1072Nt and Msed_1072Ct) or without the hexahistidine (His6)-tag (Msed_1072) was biochemically characterized using a variety of substrates over a range of temperatures and pH and in the presence of metal ions, organic solvents, and detergents. In this study, the fusion of the protein with a hexahistidine (His6)-tag did not result in a change in substrate specificity, but the findings provide information on which enzyme variant can hydrolyze fatty acid esters in the presence of various chemicals, and this has important implication for their use in industrial processes. It also demonstrates that Metallosphaera sedula Msed_1072 can have application in microalgae-based biofuel production systems.展开更多
为更好地了解勤奋金属球菌(Metallosphaera sedula)作用下沉淀的累积进程及沉淀的特性,将一株在我国首次分离鉴定的勤奋金属球菌(YN23)接种在以Fe^(2+)为能源的培养基中,于该菌最佳生长条件(pH1.5,53℃,0.2g·l_(-1)酵母提取物,30...为更好地了解勤奋金属球菌(Metallosphaera sedula)作用下沉淀的累积进程及沉淀的特性,将一株在我国首次分离鉴定的勤奋金属球菌(YN23)接种在以Fe^(2+)为能源的培养基中,于该菌最佳生长条件(pH1.5,53℃,0.2g·l_(-1)酵母提取物,30g·1_(-1)Fe_2SO_4·7H_2O and 170rpm)下培养。接种25h后,当Fe^(2+)氧化率达到90%时,开始出现沉淀,pH也达到1.92的最高值;到第95h,当沉淀累积到7.9g·1^(-1),时,沉淀反应停止,此时pH达到1.32的最低点。菌群密度随着Fe^(2+)的氧化,前期快速增长;当沉淀出现以后,随着沉淀的累积,逐渐降低。X衍射图谱、红外吸收光谱、能谱和扫描电镜数据揭示,YN23菌株合成的沉淀是黄钾铁矶和黄铵铁矾的混合物,形态特征更接近于后者。展开更多
基金financially supported by the National Natural Science Foundation of China (Nos. 31470230, 51320105006, 51604308, and 31100173)the Youth Talent Foundation of Hunan Province of China (No. 2017RS3003)the National Science Foundation of Hunan Province of China (No. 2018JJ2486)
文摘This paper describes the investigation of the secretion of extracellular polymeric substances(EPS)by an extremely thermoacidophilic archaea,Metallosphaera sedula(M.sedula),during the bioleaching of pyrite under different temperatures and discusses the relationship among the EPS secretion,its heat resistance,and its ability to bioleach pyrite.The investigation results indicate that the amount of extracellular proteins is significantly higher than the amount of extracellular polysaccharides in the extracted EPS whether free cells or attached cells;these results are quite different from the behavior of mesophilic Acidithiobacillus ferrooxidans.Although the growth of M.sedula is inhibited at 80℃,the bioleaching ability of M.sedula is only slightly lower than that at the optimum growth temperature of 72℃ because of the heat resistance mechanism based on EPS secretion.The secretion of more extracellular proteins is an important heat resistance mechanism of M.sedula.
文摘Lipid-producing microalgae are emerging as the leading platform for producing alternative biofuels in response to diminishing petroleum reserves. Optimization of fatty acid production is required for efficient conversion of microalgal fatty acids into usable transportation fuels. Microbial lipases/esterases can be used to enhance fatty acid production because of their efficacy in catalyzing hydrolysis of esters into alcohols and fatty acids while minimizing the potential poisoning of catalysts needed in the biofuel production process. Although studies have extensively focused on lipases/esterases produced by mesophilic organisms, an understanding of lipases/esterases produced by thermophilic, acidic tolerant microbes, such as Metallosphaera sedula, is limited. In this work, the carboxylesterase from Metallosphaera sedula DSM5348 encoded by Msed_1072 was recombinantly expressed in Escherichia coli strain BL21 (λDE3). The purified enzyme either with a hexahistidine (His6)-tag (Msed_1072Nt and Msed_1072Ct) or without the hexahistidine (His6)-tag (Msed_1072) was biochemically characterized using a variety of substrates over a range of temperatures and pH and in the presence of metal ions, organic solvents, and detergents. In this study, the fusion of the protein with a hexahistidine (His6)-tag did not result in a change in substrate specificity, but the findings provide information on which enzyme variant can hydrolyze fatty acid esters in the presence of various chemicals, and this has important implication for their use in industrial processes. It also demonstrates that Metallosphaera sedula Msed_1072 can have application in microalgae-based biofuel production systems.
基金Foundation item:Chinese Science Foundation for Distinguished Group(50621063)National Basic Research Program(973 Program)of P.R.China(2004CB619201)
文摘为更好地了解勤奋金属球菌(Metallosphaera sedula)作用下沉淀的累积进程及沉淀的特性,将一株在我国首次分离鉴定的勤奋金属球菌(YN23)接种在以Fe^(2+)为能源的培养基中,于该菌最佳生长条件(pH1.5,53℃,0.2g·l_(-1)酵母提取物,30g·1_(-1)Fe_2SO_4·7H_2O and 170rpm)下培养。接种25h后,当Fe^(2+)氧化率达到90%时,开始出现沉淀,pH也达到1.92的最高值;到第95h,当沉淀累积到7.9g·1^(-1),时,沉淀反应停止,此时pH达到1.32的最低点。菌群密度随着Fe^(2+)的氧化,前期快速增长;当沉淀出现以后,随着沉淀的累积,逐渐降低。X衍射图谱、红外吸收光谱、能谱和扫描电镜数据揭示,YN23菌株合成的沉淀是黄钾铁矶和黄铵铁矾的混合物,形态特征更接近于后者。