期刊
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
卷 67, 期 24, 页码 6867-6873出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.jafc.9b01856
关键词
L-DOPA; hydroxytyrosol; Escherichia coli; whole-cell catalysis; co-expression
资金
- National Key Scientific Instrument and Equipment Development Project of China [2013YQ17052504]
- Program for Changjiang Scholars and Innovative Research Team in the University of Ministry of Education of China [IRT_15R55]
- Seventh Group of Hundred-Talent Program of Shaanxi Province (2015)
- Key Project of Research and Development Plan of Shaanxi [2017ZDCXL-SF-01-02-01]
Hydroxytyrosol is a high-value-added compound with a variety of biological and pharmacological activities. In this study, a whole-cell catalytic method for the synthesis of hydroxytyrosol was developed: aromatic amino acid aminotransferase (TyrB), l-glutamate dehydrogenase (GDH), alpha-keto acid decarboxylase (PmKDC), and aldehyde reductase (YahK) were co-expressed in Escherichia coli to catalyze the synthesis of hydroxytyrosol from l-3,4-dihydroxyphenylalanine (l-DOPA). The plasmids with different copy numbers were used to balance the expression of the four enzymes, and the most appropriate strain (pRSF-yahK-tyrB and pCDF-gdh-Pmkdc) was identified. After determination of the optimum temperature (35 degrees C) and pH (7.5) for whole-cell catalysis, the yield of hydroxytyrosol reached 36.33 mM (5.59 g/L) and the space-time yield reached 0.70 g L-1 h(-1).
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