| 产碱杆菌DN25的氰降解代谢途径分析与产酶条件优化 |
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| 引用本文： | 王顺成,刘幽燕,李青云,童张法,覃益民,许建和.产碱杆菌DN25的氰降解代谢途径分析与产酶条件优化[J].化工学报,2011,62(2):482-489. |
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| 作者姓名： | 王顺成 刘幽燕 李青云 童张法 覃益民 许建和 |
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| 作者单位： | 广西大学化学化工学院；华东理工大学生物反应器工程国家重点实验室；广西生物炼制重点实验室 |
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| 摘 要： | 本实验室分离保藏的一株产碱杆菌Alcaligenes sp.DN25具有较高降氰活性,通过分析氰降解代谢产物确定了其降解途径,并根据降氰酶催化活性部位特征对产酶条件进行了优化.结果显示,氰的降解代谢途径可推断为由氰水解酶、氰水合酶和酰胺水解酶共同作用的水解途径,而其中氰水解酶的活性占主要作用;在培养基中分别添加4种含硫...
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| 关 键 词： | 产碱杆菌 氰水解酶 氰水合酶 降解代谢途径 产酶条件优化 |
Analysis of cyanide-degrading metabolism and optimization of culture condition for cyanide-degrading enzyme production from Alcaligenes sp.DN25 |
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| WANG Shuncheng,LIU Youyan,LI Qingyun,TONG Zhangfa,QIN Yimin,XU Jianhe.Analysis of cyanide-degrading metabolism and optimization of culture condition for cyanide-degrading enzyme production from Alcaligenes sp.DN25[J].Journal of Chemical Industry and Engineering(China),2011,62(2):482-489. |
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| Authors: | WANG Shuncheng LIU Youyan LI Qingyun TONG Zhangfa QIN Yimin XU Jianhe |
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| Abstract: | A strain of Alcaligenes sp., DN25, isolated and preserved in the authors’ lab, shows a high cyanide-degrading activity and thus has a potential of industrial application.The cyanide-degrading routes were determined based on the analysis of degradation products and time curves.The experiments on optimization of culture conditions for the cyanide-degrading enzyme were carried out according to the characteristics of enzyme active site.The results demonstrated that the reaction was through a hydrolytic pathway co-functioned by a cyanide hydratase, an amidase and a cyanide hydrolase, among which cyanide hydrolase played a major role.Four sulfur-containing substances were added to the medium to test the effect on specific activity of culture.DL-cysteine was found to be beneficial for the enzyme production and DL-methionine for both enzyme production and cell yield, resulting in increased specific activity of culture of 1.26 U·ml-1 and 2.10 U·ml-1，when 0.8% DL-cysteine and DL-methionine was added into fermentation culture respectively.The experiments on optimization of liquid culture conditions in 1L shake flask gave the optimal conditions as 100ml of liquid loading, pH of 9.0, inoculation size of 8%, and 0.8% DL-methionine added into the medium.Under such conditions, the specific activity of culture could reach 2.18 U·ml-1, increased by 3.46 fold compared to that before optimization.The cell growth curve had a similar trend to that of enzyme production and the stabilized phase could be reached within 40 h.The current work could lay the basis for further application and purification of the cyanide-degradating enzyme. |
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