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学科主题: 生命有机化学
题名: Properties of hybrid enzymes between synechococcus large subunits and higher plant small subunits of ribulose-1,5-bisphosphate carboxylase/oxygenase in escherichia coli
其他题名: 大肠杆菌中由巨球藻大亚基和高等植物小亚基形成杂化的核酮糖-1,5-磷酸羟化酶加氧酶的性质
作者: Wang YL(汪有力) ; Zhou JH(周佳海) ; Wang YF(王燕芳) ; Bao JS(鲍建绍) ; Chen HB(陈海宝)
通讯作者: 陈海宝
刊名: Arch. Biochem. Biophys.
发表日期: 2001
卷: 396, 期:1, 页:35-42
收录类别: SCI
部门归属: 中国科学院上海有机化学研究所生命有机国家重点实验室
英文摘要: To explore the function of small subunits of rubisco, three hydrid enzymes were synthesized in escherichia coli by construction of a transcriptionally coupled expression system in which the synthetic small subunit gene of rice, tobacco, and wheat, respectively, was cloned downstream from the large subunit gene of synechococcus sp. PCC6301. These coexpression products were detected by utilizing SDS-PAGE and confirmed by immunoblotting. The amount of carboxylase activity from the intact cells revealed that each higher plant small subunit was able to assemble with the synechococcus large subunit octamer core to form an active heterologous enzyme in E. Coli. However, in these heterologous enzymes. The interaction between large subunits and small subunits was very weak, the small subunit readily dissociated from the large subunit octamer core. A detailed kinetic assay was carried out with the partially purified hybrid enzymes. Compared to synechococcus rubisco, the activity of rice, tobacco, and whear hybrid rubisco decreased to 37, 61, and 37% of the original activity, respectively. These hybrid enzymes showed a greater affinity for CO^2 and RuBP than synechococcus rubisco. The specificity factor of the three hybrid rubiscos was 98, 84, and 76%, respectively, of the original. These results indicate for the first time that the small subunit contributes to the stability, catalytic efficiency, and CO^2/O^2 specificity of rubisco together, which suggests that small subunits may befruitful targets for engineering an improved rubisco. Meanwhile, we found that sorbitol in the culture of induced cells promoted the production of active assembled enzyme and shortened the time to reach maximal expression.
语种: 英语
相关网址: 查看原文
WOS记录号: WOS:000172578800005
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内容类型: 期刊论文
URI标识: http://ir.sioc.ac.cn/handle/331003/25749
Appears in Collections:生命有机化学国家重点实验室_期刊论文

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Recommended Citation:
Wang YL,Zhou JH,Wang YF,et al. Properties of hybrid enzymes between synechococcus large subunits and higher plant small subunits of ribulose-1,5-bisphosphate carboxylase/oxygenase in escherichia coli[J]. Arch. Biochem. Biophys.,2001,396(1):35-42.
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