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| Construction of 4-Hydroxyphenylacetate-3-hydroxylase A Expression Strain and Its Biotransformation Effect on Hydroxytyrosol |
| QIAN Xin1, GUO Hong-yan2, ZHOU Qing-feng3 |
1. Institution of Lifescience, Henan Agricultural University, Zhengzhou 450002, China;
2. Shanghai Institute of Pharmaceutical Industry, Shanghai 200040, China;
3. Institution of Lifescience, Shangqiu Normal University, Shangqiu 476000, China |
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Abstract In the present study, a recombinant strain that is able to express 4-hydroxyphenylacetate-3-hydroxylase (HHA). Hydroxytyrosol can be produced in this recombinant strain using tyrosol as the substrate. HHA gene was amplified using E. coli BL21 (DE3) as the template and then connected to the expression vector pET-28a. Obtained recombinant expression vector pET28a-HHA was transformed into competent cell BL21 (DE3). The suitable amount of tyrosol was used as raw materials to prepare hydroxytyrosol in the recombinant bacteria. Thin layer chromatography and gas chromatography were used respectively to detect the production level of hydroxytyrosol in supernatant. After the IPTG induced, two protein bands (58.8kDa and 18.5kDa) were detected by SDS-PAGE analysis. Hydroxyltyrosol was also detected by thin layer chromatography and gas chromatography. The results showed that the expression recombinant strain HHA was successfully conducted and this strain can effectively convert tyrosol to hydroxytyrosol.
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Received: 22 December 2014
Published: 25 March 2015
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