PMID- 33418890
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20210218
IS  - 2079-7737 (Print)
IS  - 2079-7737 (Electronic)
IS  - 2079-7737 (Linking)
VI  - 10
IP  - 1
DP  - 2021 Jan 6
TI  - Discovery and Validation of a Novel Step Catalyzed by OsF3H in the Flavonoid 
      Biosynthesis Pathway.
LID - 10.3390/biology10010032 [doi]
LID - 32
AB  - Kaempferol and quercetin are the essential plant secondary metabolites that 
      confer huge biological functions in the plant defense system. In this study, 
      biosynthetic pathways for kaempferol and quercetin were constructed in 
      Saccharomyces cerevisiae using naringenin as a substrate. OsF3H was cloned into 
      pRS42K yeast episomal plasmid (YEp) vector and the activity of the target gene 
      was analyzed in engineered and empty strains. We confirmed a novel step of 
      kaempferol and quercetin biosynthesis directly from naringenin, catalyzed by the 
      rice flavanone 3-hydroxylase (F3H). The results were confirmed through thin layer 
      chromatography (TLC) followed by western blotting, nuclear magnetic resonance 
      (NMR), and liquid chromatography-mass spectrometry LCMS-MS. TLC showed positive 
      results when comparing both compounds extracted from the engineered strain with 
      the standard reference. Western blotting confirmed the lack of OsF3H activity in 
      empty strains and confirmed high OsF3H expression in engineered strains. NMR 
      spectroscopy confirmed only quercetin, while LCMS-MS results revealed that F3H is 
      responsible for the conversion of naringenin to both kaempferol and quercetin.
FAU - Jan, Rahmatullah
AU  - Jan R
AUID- ORCID: 0000-0001-5788-3017
AD  - Division of Plant Biosciences, School of Applied Biosciences, College of 
      Agriculture & Life Science, Kyungpook National University, 80 Dahak-ro, Buk-gu, 
      Daegu 41566, Korea.
FAU - Asaf, Sajjad
AU  - Asaf S
AUID- ORCID: 0000-0001-8216-7549
AD  - Natural and Medical Science Research Center, University of Nizwa 616, Nizwa 611, 
      Oman.
FAU - Paudel, Sanjita
AU  - Paudel S
AD  - College of Pharmacy, Research Institute of Pharmaceutical Sciences, Kyungpook 
      National University, 80 Dahak-ro, Buk-gu, Daegu 41566, Korea.
FAU - Lubna
AU  - Lubna
AD  - Department of Botany, Garden Campus, Abdul Wali Khan University, Mardan 23200, 
      Pakistan.
FAU - Lee, Sangkyu
AU  - Lee S
AD  - College of Pharmacy, Research Institute of Pharmaceutical Sciences, Kyungpook 
      National University, 80 Dahak-ro, Buk-gu, Daegu 41566, Korea.
FAU - Kim, Kyung-Min
AU  - Kim KM
AUID- ORCID: 0000-0003-4812-6297
AD  - Division of Plant Biosciences, School of Applied Biosciences, College of 
      Agriculture & Life Science, Kyungpook National University, 80 Dahak-ro, Buk-gu, 
      Daegu 41566, Korea.
LA  - eng
GR  - PJ014793012020/Rural Development Administration/
PT  - Journal Article
DEP - 20210106
PL  - Switzerland
TA  - Biology (Basel)
JT  - Biology
JID - 101587988
PMC - PMC7825110
OTO - NOTNLM
OT  - hydroxylation
OT  - kaempferol
OT  - naringenin
OT  - nuclear magnetic resonance
OT  - yeast episomal plasmid
COIS- The authors declare no conflict of interest.
EDAT- 2021/01/10 06:00
MHDA- 2021/01/10 06:01
PMCR- 2021/01/06
CRDT- 2021/01/09 01:01
PHST- 2020/11/23 00:00 [received]
PHST- 2020/12/30 00:00 [revised]
PHST- 2021/01/01 00:00 [accepted]
PHST- 2021/01/09 01:01 [entrez]
PHST- 2021/01/10 06:00 [pubmed]
PHST- 2021/01/10 06:01 [medline]
PHST- 2021/01/06 00:00 [pmc-release]
AID - biology10010032 [pii]
AID - biology-10-00032 [pii]
AID - 10.3390/biology10010032 [doi]
PST - epublish
SO  - Biology (Basel). 2021 Jan 6;10(1):32. doi: 10.3390/biology10010032.