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短密青霉NRRL 864异戊烯基转移酶PbPT的克隆表达及功能分析
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作者 孙靖然 刘长宁 +2 位作者 李荣贵 张伟 李盛英 《青岛大学学报(工程技术版)》 CAS 2016年第2期107-115,共9页
在对丝状真菌短密青霉(penicillium brevicompactum NRRL 864)的全基因组进行分析时,发现该真菌可能表达一种异戊烯基转移酶PbPT。通过反转录RT-PCR(Reverse Transcription,RT-PCR)获得真菌cDNA,并以cDNA为模版,将编码PbPT的开放阅读框... 在对丝状真菌短密青霉(penicillium brevicompactum NRRL 864)的全基因组进行分析时,发现该真菌可能表达一种异戊烯基转移酶PbPT。通过反转录RT-PCR(Reverse Transcription,RT-PCR)获得真菌cDNA,并以cDNA为模版,将编码PbPT的开放阅读框克隆到pET-28b构建表达载体pET28b-PbPT,将pET28b-PbPT转化Escherichia coli BL21(DE3)获得表达菌株,异丙基-β-D-硫代半乳糖苷(Isopropyl-β-D-thiogalactoside,IPTG)诱导高效表达PbPT,利用Ni-NTA树脂亲和纯化重组PbPT活性蛋白,并对该蛋白进行了底物特异性分析、产物结构鉴定以及酶促动力学分析。分析结果表明,以二甲基丙烯基二磷酸(dimethylallyl pyrophosphate,DMAPP)为供体,PbPT特异性催化异戊烯基转移至Brevianamide F的α位C上,生成Deoxybrevianamide E。Brevianamide F是合成真菌生物碱类杀虫抗生素brevianamides的重要中间体,真菌吲哚类异戊烯基转移酶家族新成员PbPT的发现。该研究为进一步寻找和阐明真菌生物碱类杀虫抗生素brevianamides的生物合成基因簇和合成途径奠定了基础。 展开更多
关键词 短密青霉 异戊烯基转移酶 丝状真菌 dmapp 功能分析
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Corrigendum to"Complete biosynthesis of the potential medicine icaritin by engineered Saccharomyces cerevisiae and Escherichia coli"[Sci.Bull.66(18)(2021)1906-1916]
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作者 Pingping Wang Chaojing Li +8 位作者 Xiaodong Li Wenjun Huang Yan Wang Jiali Wang Yanjun Zhang Xiaoman Yang Xing Yan Ying Wang Zhihua Zhou 《Science Bulletin》 2025年第11期1891-1891,共1页
The authors would like to apologize that,due to their neglect,some mistakes in the"2.Materials and methods"section need to be corrected.(1)The unit of DMAPP and flavonoid substrates in the“2.3.Enzymatic ass... The authors would like to apologize that,due to their neglect,some mistakes in the"2.Materials and methods"section need to be corrected.(1)The unit of DMAPP and flavonoid substrates in the“2.3.Enzymatic assays of prenyltransferases"section was not correctly written.The original description“200 mmol/L dimethylallyl pyrophosphate(DMAPP),200 mmol/L naringenin,kaempferol,or kaempferide as substrate"should be corrected to“200μmol/L dimethylallyl pyrophosphate(DMAPP),200μmol/L naringenin,kaempferol,or kaempferide as substrate"(2)The concentration of acetate acid in“2.8.Chemical analysis"was not correctly written.The original description“0.01%acetate acid(A)"should be corrected to“0.1%acetate acid(A)".The authors declare that these corrections do not change the results or conclusions of this paper.The authors sincerely apologize for this omission. 展开更多
关键词 BIOSYNTHESIS Escherichia coli flavonoid substrates Saccharomyces cerevisiae enzymatic assays dimethylallyl pyrophosphate dmapp mol l ICARITIN correction
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