Among 39 species of microbial strains,Rhodococcus sp.AS 4.1147 possessed the ability to selectively acetylate puerarin（1） at C-6 position of the glucosyl moiety.The structure of the acetylated product,6’’-O-acetyl...Among 39 species of microbial strains,Rhodococcus sp.AS 4.1147 possessed the ability to selectively acetylate puerarin（1） at C-6 position of the glucosyl moiety.The structure of the acetylated product,6’’-O-acetylpuerarin（2） was determined by the analysis of MS,NMR spectroscopic data.The isolated yield of 2 was 22.2%.展开更多
为解决乙草胺残留问题,采用连续传代富集培养的方法,从长期生产乙草胺的农药厂污染土壤中分离筛选到1株乙草胺降解菌株AC-1。经过形态学特征、生理生化特征和16S rRNA基因序列系统发育分析,菌株AC-1被鉴定为红球菌属(Rhodococcus)。菌株...为解决乙草胺残留问题,采用连续传代富集培养的方法,从长期生产乙草胺的农药厂污染土壤中分离筛选到1株乙草胺降解菌株AC-1。经过形态学特征、生理生化特征和16S rRNA基因序列系统发育分析,菌株AC-1被鉴定为红球菌属(Rhodococcus)。菌株AC-1在48h内能将0.2 m M的乙草胺完全降解,但不能矿化乙草胺。借助LC-MS,确定乙草胺降解终产物为2-氯-N-(2-乙基-6-甲基苯基)乙酰胺(CMEPA)。菌株AC-1降解乙草胺的最适温度为30℃,最适p H值为7.5。0.1 m M的Cu2+和Hg2+对菌株AC-1降解乙草胺具有很强的抑制作用,而0.1 m M的Fe2+则对菌株AC-1降解乙草胺具有微弱的促进作用。菌株AC-1对乙草胺的降解效果与起始接种量呈正相关。菌株AC-1对甲草胺和丁草胺表现出良好的降解效果,但对异丙甲草胺的降解能力非常微弱。土壤降解试验表明,投加菌株AC-1可以明显促进土壤中乙草胺的降解。本研究为乙草胺的有效降解提供了数据和技术支持。展开更多
二苯并噻吩(DBT)及其衍生物微生物脱硫的4S途径需要4个酶(DszA,DszB,DszC and DszD)参与催化。其中DBT单加氧酶(DszC or DBT-MO)和DBT-砜单加氧酶(DszA or DBTO2-MO)都是黄素依赖型氧化酶,它们的催化反应需要菌体中还原型的黄素单核苷酸...二苯并噻吩(DBT)及其衍生物微生物脱硫的4S途径需要4个酶(DszA,DszB,DszC and DszD)参与催化。其中DBT单加氧酶(DszC or DBT-MO)和DBT-砜单加氧酶(DszA or DBTO2-MO)都是黄素依赖型氧化酶,它们的催化反应需要菌体中还原型的黄素单核苷酸(FMNH2),FMNH2由辅酶黄素还原酶(DszD)再生。因此,共表达DszA,DszB,DszC和DszD可以提高整个脱硫途径的速率。构建了两个不相容性表达载体pBADD和paN2并在大肠杆菌中实现了4个脱硫酶基因的共表达。DszA,DszB,DszC和DszD的可溶性蛋白表达量分别占菌体总蛋白质的7.6%,3.5%,3.1%和18%。共表达时的脱硫活性是单独用paN2表达时的5.4倍,并对工程菌休止细胞脱除模拟柴油中DBT的活性进行了研究。展开更多
Inducing expression and the reaction characteristic of nitrile hydratase （NHase） from Rhodococcus sp. SHZ-1 were investigated. The results showed that the expression of NHase was greatly enhanced with the cooperatio...Inducing expression and the reaction characteristic of nitrile hydratase （NHase） from Rhodococcus sp. SHZ-1 were investigated. The results showed that the expression of NHase was greatly enhanced with the cooperation of acrylonitrile and ammonium chloride as inducer in the medium and the specific activity of NHase was increased of 44%. Then the temperature, pH, concentration of acrylonitrile and acrylamide were evaluated, which affected the activity and reaction characteristic of NHase. It was found that the temperature and concentration of acrylarnide were the most important factors for the catalyzation of NHase. The optimal catalysis temperature of NHase from Rhodococcus sp. SHZ-1 was 30℃, and the activation energy of the hydration of NHase was 90.2kJ·mol^-1 in the temperature range from 5℃ to 30℃. Kmof NHase was 0.095mol·L^-1 using acrylonitrile（AN） as substrate, and NHase activity was inhibited seriously when acrylonitrile concentration was up to 40g·L^-1, the substrate inhibition constant Ki is 0.283mol·L^-1. Moreover, the NHase from Rhodococcus sp. SHZ-1 had very strong tolerance to acrylamide, in which the final concentration of acrylamide reached to 642g·L^-1 and the residual activity of NHase still maintained 8.6% of the initial enzyme activity.展开更多
The biodegradation characteristic and potential metabolic pathway for removal of environmental N,N-dimethylacetamide （DMAC） by Rhodococcus sp. strain B83 was studied. Rhodococcus sp. strain B83 was isolated from the...The biodegradation characteristic and potential metabolic pathway for removal of environmental N,N-dimethylacetamide （DMAC） by Rhodococcus sp. strain B83 was studied. Rhodococcus sp. strain B83 was isolated from the rhizosphere of a pagoda tree and proved capable of utilizing DMAC as sole source of carbon and nitrogen. Batch culture studies showed that strain B83 could tolerate up to 25 g/L DMAC and showed distinct growth on possible catabolic intermediates except for acetate. The nitrogen balance analysis revealed that approximately 71% of the initial nitrogen was converted to organic nitrogen. DMAC degradation has led to accumulation of acetate and organic nitrogen, meanwhile traces of nitrate and ammonia was build-up but without nitrite. The growth of strain B83 could be inhibited by adding exogenous acetate. By means of the assay of enzymatic degradation of DMAC, several catabolic intermediates at different intervals were observed and identified. Based on the results obtained from culture solution and enzymatic degradation assay, a detailed pathway is proposed for DMAC biodegradation.展开更多
基金The Science & Technology Project of Guangdong Province (Grant No. 2011A080403020)the Program for New Century Excellent Talents in University (Grant No. NCET-06-0155)
文摘Among 39 species of microbial strains,Rhodococcus sp.AS 4.1147 possessed the ability to selectively acetylate puerarin（1） at C-6 position of the glucosyl moiety.The structure of the acetylated product,6’’-O-acetylpuerarin（2） was determined by the analysis of MS,NMR spectroscopic data.The isolated yield of 2 was 22.2%.
文摘为解决乙草胺残留问题,采用连续传代富集培养的方法,从长期生产乙草胺的农药厂污染土壤中分离筛选到1株乙草胺降解菌株AC-1。经过形态学特征、生理生化特征和16S rRNA基因序列系统发育分析,菌株AC-1被鉴定为红球菌属(Rhodococcus)。菌株AC-1在48h内能将0.2 m M的乙草胺完全降解,但不能矿化乙草胺。借助LC-MS,确定乙草胺降解终产物为2-氯-N-(2-乙基-6-甲基苯基)乙酰胺(CMEPA)。菌株AC-1降解乙草胺的最适温度为30℃,最适p H值为7.5。0.1 m M的Cu2+和Hg2+对菌株AC-1降解乙草胺具有很强的抑制作用,而0.1 m M的Fe2+则对菌株AC-1降解乙草胺具有微弱的促进作用。菌株AC-1对乙草胺的降解效果与起始接种量呈正相关。菌株AC-1对甲草胺和丁草胺表现出良好的降解效果,但对异丙甲草胺的降解能力非常微弱。土壤降解试验表明,投加菌株AC-1可以明显促进土壤中乙草胺的降解。本研究为乙草胺的有效降解提供了数据和技术支持。
文摘二苯并噻吩(DBT)及其衍生物微生物脱硫的4S途径需要4个酶(DszA,DszB,DszC and DszD)参与催化。其中DBT单加氧酶(DszC or DBT-MO)和DBT-砜单加氧酶(DszA or DBTO2-MO)都是黄素依赖型氧化酶,它们的催化反应需要菌体中还原型的黄素单核苷酸(FMNH2),FMNH2由辅酶黄素还原酶(DszD)再生。因此,共表达DszA,DszB,DszC和DszD可以提高整个脱硫途径的速率。构建了两个不相容性表达载体pBADD和paN2并在大肠杆菌中实现了4个脱硫酶基因的共表达。DszA,DszB,DszC和DszD的可溶性蛋白表达量分别占菌体总蛋白质的7.6%,3.5%,3.1%和18%。共表达时的脱硫活性是单独用paN2表达时的5.4倍,并对工程菌休止细胞脱除模拟柴油中DBT的活性进行了研究。
基金Supported by the National Natural Science Foundation of China （No.20466002）, the Program for New Century Excellent Talents in University （NCET-04-089） and the Key Research Projects in the Uygur Autonomous Region of Xinjiang （No.200332109）.
文摘Inducing expression and the reaction characteristic of nitrile hydratase （NHase） from Rhodococcus sp. SHZ-1 were investigated. The results showed that the expression of NHase was greatly enhanced with the cooperation of acrylonitrile and ammonium chloride as inducer in the medium and the specific activity of NHase was increased of 44%. Then the temperature, pH, concentration of acrylonitrile and acrylamide were evaluated, which affected the activity and reaction characteristic of NHase. It was found that the temperature and concentration of acrylarnide were the most important factors for the catalyzation of NHase. The optimal catalysis temperature of NHase from Rhodococcus sp. SHZ-1 was 30℃, and the activation energy of the hydration of NHase was 90.2kJ·mol^-1 in the temperature range from 5℃ to 30℃. Kmof NHase was 0.095mol·L^-1 using acrylonitrile（AN） as substrate, and NHase activity was inhibited seriously when acrylonitrile concentration was up to 40g·L^-1, the substrate inhibition constant Ki is 0.283mol·L^-1. Moreover, the NHase from Rhodococcus sp. SHZ-1 had very strong tolerance to acrylamide, in which the final concentration of acrylamide reached to 642g·L^-1 and the residual activity of NHase still maintained 8.6% of the initial enzyme activity.
基金supported by the National Natural Science Foundation of China(No.51308440)the Natural Science Foundation of Shaanxi Province(No.2014JM7260)Shaanxi Provincial Department of Education Fund(No.15JK1439)are gratefully acknowledged
文摘The biodegradation characteristic and potential metabolic pathway for removal of environmental N,N-dimethylacetamide （DMAC） by Rhodococcus sp. strain B83 was studied. Rhodococcus sp. strain B83 was isolated from the rhizosphere of a pagoda tree and proved capable of utilizing DMAC as sole source of carbon and nitrogen. Batch culture studies showed that strain B83 could tolerate up to 25 g/L DMAC and showed distinct growth on possible catabolic intermediates except for acetate. The nitrogen balance analysis revealed that approximately 71% of the initial nitrogen was converted to organic nitrogen. DMAC degradation has led to accumulation of acetate and organic nitrogen, meanwhile traces of nitrate and ammonia was build-up but without nitrite. The growth of strain B83 could be inhibited by adding exogenous acetate. By means of the assay of enzymatic degradation of DMAC, several catabolic intermediates at different intervals were observed and identified. Based on the results obtained from culture solution and enzymatic degradation assay, a detailed pathway is proposed for DMAC biodegradation.
