Conversion of fatty aldehydes into alk (a/e)nes by in vitro reconstituted cyanobacterial aldehyde-deformylating oxygenase with the cognate electron transfer system

Biotechnology for Biofuels

Jingjing Zhang¹  Xuefeng Lu²  Jian-Jun Li² 

[1] University of Chinese Academy of Sciences, Beijing, 100049, China

[2] Key Laboratory of Biofuels, Shandong Provincial Key Laboratory of Energy Genetics, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, No. 189 Songling Road, Qingdao, 266101, China

关键词: The cognate reducing system;    Ferredoxin-NADP⁺ reductase;    Ferredoxin;    Aldehyde-deformylating oxygenase;    Synechococcus elongatus PCC7942;    Fatty alk(a/e)ne;    Biofuels;   

Others  :  798018
DOI  :  10.1186/1754-6834-6-86

 received in 2013-02-04, accepted in 2013-06-03,  发布年份 2013

【 摘 要 】

Background

Biosynthesis of fatty alk(a/e)ne in cyanobacteria has been considered as a potential basis for the sunlight-driven and carbon-neutral bioprocess producing advanced solar biofuels. Aldehyde-deformylating oxygenase (ADO) is a key enzyme involved in that pathway. The heterologous or chemical reducing systems were generally used in in vitro ADO activity assay. The cognate electron transfer system from cyanobacteria to support ADO activity is still unknown.

Results

We identified the potential endogenous reducing system including ferredoxin (Fd) and ferredoxin-NADP⁺ reductase (FNR) to support ADO activity in Synechococcus elongatus PCC7942. ADO (Synpcc7942_1593), FNR (SynPcc7942_0978), and Fd (SynPcc7942_1499) from PCC7942 were cloned, overexpressed, purified, and characterized. ADO activity was successfully supported with the endogenous electron transfer system, which worked more effectively than the heterologous and chemical ones. The results of the hybrid Fd/FNR reducing systems demonstrated that ADO was selective against Fd. And it was observed that the cognate reducing system produced less H₂O₂ than the heterologous one by 33% during ADO-catalyzed reactions. Importantly, k_cat value of ADO 1593 using the homologous Fd/FNR electron transfer system is 3.7-fold higher than the chemical one.

Conclusions

The cognate electron transfer system from cyanobacteria to support ADO activity was identified and characterized. For the first time, ADO was functionally in vitro reconstituted with the endogenous reducing system from cyanobacteria, which supported greater activity than the surrogate and chemical ones, and produced less H₂O₂ than the heterologous one. The identified Fd/FNR electron transfer system will be potentially useful for improving ADO activity and further enhancing the biosynthetic efficiency of hydrocarbon biofuels in cyanobacteria.

【 授权许可】

   
2013 Zhang et al.; licensee BioMed Central Ltd.

【 图 表 】

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