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在含有化学计量的大亚基和小亚基的大肠杆菌中活性蓝藻1,5-二磷酸核酮糖羧化酶/加氧酶的表达与组装

Expression and assembly of active cyanobacterial ribulose-1,5-bisphosphate carboxylase/oxygenase in Escherichia coli containing stoichiometric amounts of large and small subunits.

作者信息

Tabita F R, Small C L

出版信息

Proc Natl Acad Sci U S A. 1985 Sep;82(18):6100-3. doi: 10.1073/pnas.82.18.6100.

DOI:10.1073/pnas.82.18.6100
PMID:3929249
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC390707/
Abstract

The genes for the large and small subunits of ribulose-1,5-bisphosphate carboxylase/oxygenase from the cyanobacterium Anacystis nidulans were subcloned into plasmid pUC9. After induction, both genes were expressed in Escherichia coli and the subunits were assembled into an active holoenzyme. The enzyme was purified from E. coli to high specific activity and was found to contain equimolar amounts of large and small subunits. The assembly of the hexadecameric ribulose bisphosphate carboxylase/oxygenase in E. coli should provide the basis for studies on the mechanism of assembly and the role of small subunits in catalysis.

摘要

将来自蓝藻集胞藻6803的1,5-二磷酸核酮糖羧化酶/加氧酶的大亚基和小亚基基因亚克隆到质粒pUC9中。诱导后，这两个基因在大肠杆菌中表达，并且亚基组装成活性全酶。该酶从大肠杆菌中纯化至高比活性，发现含有等摩尔量的大亚基和小亚基。在大肠杆菌中十六聚体1,5-二磷酸核酮糖羧化酶/加氧酶的组装应为研究组装机制和小亚基在催化中的作用提供基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0461/390707/c4f021d25a79/pnas00358-0086-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0461/390707/c4f021d25a79/pnas00358-0086-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0461/390707/c4f021d25a79/pnas00358-0086-a.jpg

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1
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本文引用的文献

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Active-site carbamate formation and reaction-intermediate-analog binding by ribulosebisphosphate carboxylase/oxygenase in the absence of its small subunits.在缺乏其小亚基的情况下，核酮糖二磷酸羧化酶/加氧酶的活性部位氨基甲酰化形成和反应中间类似物结合。
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Active-site histidines in recombinant cyanobacterial ribulose-1,5-bisphosphate carboxylase/oxygenase examined by site-directed mutagenesis.通过定点突变研究重组蓝藻核酮糖-1,5-二磷酸羧化酶/加氧酶的活性部位组氨酸。
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Synthesis of active Olisthodiscus luteus ribulose-1,5-bisphosphate carboxylase in Escherichia coli.在大肠杆菌中合成活性的 Olisthodiscus luteus 核酮糖-1,5-二磷酸羧化酶。
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A Rubisco mutant that confers growth under a normally "inhibitory" oxygen concentration.一种在正常“抑制性”氧浓度下仍能实现生长的核酮糖-1,5-二磷酸羧化酶/加氧酶突变体。
Biochemistry. 2009 Sep 29;48(38):9076-83. doi: 10.1021/bi9006385.
在蓝细菌集胞藻6301中，1,5 - 二磷酸核酮糖羧化酶/加氧酶小亚基的基因位于大亚基基因附近。
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Ribulose 1,5-bisphosphate carboxylase from the halophilic cyanobacterium Aphanothece halophytica.来自嗜盐蓝藻植物盐生隐球藻的1,5-二磷酸核酮糖羧化酶。
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The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers.pUC质粒，一种源自M13mp7的用于插入诱变和使用合成通用引物进行测序的系统。
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