Development of an activity-directed selection system enabled significant improvement of the carboxylation efficiency of Rubisco
文献类型：期刊论文
| 作者 | Cai, Zhen1; Liu, Guoxia1; Zhang, Junli2; Li, Yin1 |
| 刊名 | PROTEIN & CELL
 |
| 出版日期 | 2014-07-01 |
| 卷号 | 5期号:7页码:552-562 |
| 关键词 | carboxylation efficiency CO2 fixation directed evolution Rubisco Synechococcus sp PCC7002 |
| 英文摘要 | Photosynthetic CO2 fixation is the ultimate source of organic carbon on earth and thus is essential for crop production and carbon sequestration. Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) catalyzes the first step of photosynthetic CO2 fixation. However, the extreme low carboxylation efficiency of Rubisco makes it the most attractive target for improving photosynthetic efficiency. Extensive studies have focused on re-engineering a more efficient enzyme, but the effort has been impeded by the limited understanding of its structure-function relationships and the lack of an efficient selection system towards its activity. To address the unsuccessful molecular engineering of Rubisco, we developed an Escherichia coli-based activity-directed selection system which links the growth of host cell solely to the Rubisco activity therein. A Synechococcus sp. PCC7002 Rubisco mutant with E49V and D82G substitutions in the small subunit was selected from a total of 15,000 mutants by one round of evolution. This mutant showed an 85% increase in specific carboxylation activity and a 45% improvement in catalytic efficiency towards CO2. The small-subunit E49V mutation was speculated to influence holoenzyme catalysis through interaction with the large-subunit Q225. This interaction is conserved among various Rubisco from higher plants and Chlamydomonas reinhardtii. Knowledge of these might provide clues for engineering Rubisco from higher plants, with the potential of increasing the crop yield. |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
| 类目[WOS] | Cell Biology |
| 研究领域[WOS] | Cell Biology |
| 关键词[WOS] | RIBULOSE-BISPHOSPHATE CARBOXYLASE ; PLANT SMALL SUBUNITS ; RIBULOSE-1,5-BISPHOSPHATE CARBOXYLASE/OXYGENASE ; CATALYTIC EFFICIENCY ; CO2/O-2 SPECIFICITY ; THERMAL-STABILITY ; 1,5-BISPHOSPHATE CARBOXYLASE ; SPECTROPHOTOMETRIC ASSAY ; HEXADECAMERIC RUBISCO ; CRYSTAL-STRUCTURE |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000339333400008 |
| 源URL | [http://124.16.173.210/handle/834782/2268]  |
| 专题 | 天津工业生物技术研究所_总体研究部_期刊论文 |
| 作者单位 | 1.Chinese Acad Sci, Inst Microbiol, CAS Key Lab Microbial Physiol & Metab Engn, Beijing 100101, Peoples R China 2.Chinese Acad Sci, Tianjin Inst Ind Biotechnol, Tianjin 300308, Peoples R China |
| 推荐引用方式 GB/T 7714 | Cai, Zhen,Liu, Guoxia,Zhang, Junli,et al. Development of an activity-directed selection system enabled significant improvement of the carboxylation efficiency of Rubisco[J]. PROTEIN & CELL,2014,5(7):552-562. |
| APA | Cai, Zhen,Liu, Guoxia,Zhang, Junli,&Li, Yin.(2014).Development of an activity-directed selection system enabled significant improvement of the carboxylation efficiency of Rubisco.PROTEIN & CELL,5(7),552-562. |
| MLA | Cai, Zhen,et al."Development of an activity-directed selection system enabled significant improvement of the carboxylation efficiency of Rubisco".PROTEIN & CELL 5.7(2014):552-562. |
入库方式： OAI收割
来源：天津工业生物技术研究所
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