Metabolic engineering of Escherichia coli for the production of isoflavonoid-4'-O-methoxides and their biological activities.
Isoflavonoid representatives such as genistein and daidzein are highly potent anticancer, antibacterial, and antioxidant agents. It have been demonstrated that methylation of flavonoids enhanced the transporting ability, which lead to facilitated absorption and greatly increased bioavailability. In this paper, genetically engineered Escherichia coli was reconstructed by harboring E. coli K12-derived metK encoding S-adenosine-l-methionine (SAM) synthase (accession number: K02129) for enhancement of SAM as a precursor and Streptomyces avermitilis originated SaOMT2 (O-methyltransferase, accession number: NP_823558) for methylation of daidzein and genistein as preferred substrates. The formation of desired products via biotransformation including 4'-O-methyl-genistein and 4'-O-methyl-daidzein was confirmed individually by using chromatographical methods such as high-performance liquid chromatography, liquid chromatography/time-of-flight/mass spectrometry (LC-TOF-MS), and nuclear magnetic resonance (NMR), and NMR (1H and13C). Furthermore, substrates concentration, incubation time, and media parameters were optimized using flask culture. Finally, the most fit conditions were applied for fed-batch fermentation with scale-up to 3 L (working volume) to obtain the maximum yield of the products including 164.25 µM (46.81 mg/L) and 382.50 µM (102.88 mg/L) for 4'-O-methyl genistein and 4'-O-methyl daidzein, respectively. In particular, potent inhibitory activities of those isoflavonoid methoxides against the growth of cancer line (B16F10, AGS, and HepG2) and human umbilical vein endothelial cells were investigated and demonstrated. Taken together, this research work described the production of isoflavonoid-4'-O-methoxides by E. coli engineering, improvement of production, characterization of produced compounds, and preliminary in vitro biological activities of the flavonoids being manufactured.
Department of BT-Convergent Pharmaceutical Engineering, Sun Moon University, Asansi, Chungnam, Republic of Korea.
Niranjan
N
Koirala
Department of BT-Convergent Pharmaceutical Engineering, Sun Moon University, Asansi, Chungnam, Republic of Korea.|Department of Life Science and Biochemical Engineering, Sun Moon University, Asansi, Chungnam, Republic of Korea.
Ramesh Prasad
RP
Pandey
Center for Molecular Biology, Institute of Research and Development, Duy Tan University, Danang city, Vietnam.
Nguyen Huy
NH
Thuan
Department of BT-Convergent Pharmaceutical Engineering, Sun Moon University, Asansi, Chungnam, Republic of Korea.
Gopal Prasad
GP
Ghimire
Department of BT-Convergent Pharmaceutical Engineering, Sun Moon University, Asansi, Chungnam, Republic of Korea.|Department of Life Science and Biochemical Engineering, Sun Moon University, Asansi, Chungnam, Republic of Korea.
Hye Jin
HJ
Jung
Department of BT-Convergent Pharmaceutical Engineering, Sun Moon University, Asansi, Chungnam, Republic of Korea.|Department of Life Science and Biochemical Engineering, Sun Moon University, Asansi, Chungnam, Republic of Korea.
Tae-Jin
TJ
Oh
Department of BT-Convergent Pharmaceutical Engineering, Sun Moon University, Asansi, Chungnam, Republic of Korea.|Department of Life Science and Biochemical Engineering, Sun Moon University, Asansi, Chungnam, Republic of Korea.
Jae Kyung
JK
Sohng
D000970:Antineoplastic Agents / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology*; D049109:Cell Proliferation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004354:Drug Screening Assays, Antitumor; D004926:Escherichia coli / Q000737:chemistry / Q000378:metabolism*; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects; D006801:Humans; D007529:Isoflavones / Q000096:biosynthesis* / Q000737:chemistry / Q000494:pharmacology*; D060847:Metabolic Engineering; D000432:Methanol / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D015394:Molecular Structure; D013329:Structure-Activity Relationship; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; D007529:Isoflavones; D000432:Methanol
biosynthesis; cancer; isoflavonoid-4′-O-methoxides; methylation