Discovery of Five New Ethylene-Forming Enzymes for Clean Production of Ethylene in E. coli

Autor: Yixuan Cui, Ying Jiang, Meng Xiao, Muhammad Zeeshan Munir, Sadaf Riaz, Faiz Rasul, Maurycy Daroch
Jazyk: angličtina
Rok vydání: 2022
Předmět:
Zdroj: International Journal of Molecular Sciences, Vol 23, Iss 9, p 4500 (2022)
Druh dokumentu: article
ISSN: 1422-0067
1661-6596
DOI: 10.3390/ijms23094500
Popis: Ethylene is an essential platform chemical with a conjugated double bond, which can produce many secondary chemical products through copolymerisation. At present, ethylene production is mainly from petroleum fractionation and cracking, which are unsustainable in the long term, and harmful to our environment. Therefore, a hot research field is seeking a cleaner method for ethylene production. Based on the model ethylene-forming enzyme (Efe) AAD16440.1 (6vp4.1.A) from Pseudomonas syringae pv. phaseolicol, we evaluated five putative Efe protein sequences using the data derived from phylogenetic analyses and the conservation of their catalytic structures. Then, pBAD expression frameworks were constructed, and relevant enzymes were expressed in E. coli BL21. Finally, enzymatic activity in vitro and in vivo was detected to demonstrate their catalytic activity. Our results show that the activity in vitro measured by the conversion of α-ketoglutarate was from 0.21–0.72 μmol ethylene/mg/min, which varied across the temperatures. In cells, the activity of the new Efes was 12.28–147.43 μmol/gDCW/h (DCW, dry cellular weight). Both results prove that all the five putative Efes could produce ethylene.
Databáze: Directory of Open Access Journals
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