期刊
ACS SUSTAINABLE CHEMISTRY & ENGINEERING
卷 8, 期 14, 页码 5558-5565出版社
AMER CHEMICAL SOC
DOI: 10.1021/acssuschemeng.9b07457
关键词
Linoleic acid; Biotransformation; 13S-Hydroxyoctadecadienoic acid; Lipoxygenase; Cyanobacteria
资金
- National Natural Science Foundation of China [21536004, 21776085, 21871085]
- Natural Science Foundation of Shanghai [18ZR1408400]
- National Key Research and Development Program of China [2018YFC1706200, 2019YFA09005000]
- Fundamental Research Funds for the Central Universities [22221818014]
Enzymatic dioxygenation of unsaturated fatty acids by lipoxygenases (LOXs) represents a useful method in the transformation of bioresources for hydroperoxy and hydroxy fatty acids production. Difficulties in screening for optimal enzymes and in producing LOXs in sufficient quantity limit the large-scale application of LOXs. In this work, we identified three putative LOXs from Cyanobacteria, i.e., Ca-LOX from Calothrix sp. HK-06, Ri-LOX from Rivularia sp. PCC 7116, and Tb-LOX from Tolypothrix bouteillei VB521301. After heterologous expression in Escherichia coli and purification by HisTrap affinity chromatography, these LOXs showed high specific activities toward linoleic acid (73.1, 68.8, and 10.1 U mg(-1)protein, respectively). All three enzymes were identified as linoleate 13S-lipoxygenases. Ca-LOX and Ri-LOX are the highest-activity LOXs yet reported, and Ri-LOX gave a high titer of 38.3 U/mL on heterologous expression in E. coli. Consequently, by using Ri-LOX as an efficient biocatalyst with a subsequent reduction step, the renewable material linoleic acid was efficiently transformed into a value-added chemical, 13(S)-hydroxy-9,11-(Z,E)-octadecadienoic acid (13S-HODE), with an unprecedented space-time yield of 1008 g L-1 d(-1), indicating great potential of this LOX for sustainable production of 13-HODE and its derivatives.
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