PMID- 34583991
OWN - NLM
STAT- MEDLINE
DCOM- 20211214
LR  - 20220329
IS  - 1091-6490 (Electronic)
IS  - 0027-8424 (Print)
IS  - 0027-8424 (Linking)
VI  - 118
IP  - 40
DP  - 2021 Oct 5
TI  - FAD-dependent C-glycoside-metabolizing enzymes in microorganisms: Screening, 
      characterization, and crystal structure analysis.
LID - 10.1073/pnas.2106580118 [doi]
LID - e2106580118
AB  - C-glycosides have a unique structure, in which an anomeric carbon of a sugar is 
      directly bonded to the carbon of an aglycone skeleton. One of the natural 
      C-glycosides, carminic acid, is utilized by the food, cosmetic, and 
      pharmaceutical industries, for a total of more than 200 tons/y worldwide. 
      However, a metabolic pathway of carminic acid has never been identified. In this 
      study, we isolated the previously unknown carminic acid-catabolizing 
      microorganism and discovered a flavoenzyme "C-glycoside 3-oxidase" named CarA 
      that catalyzes oxidation of the sugar moiety of carminic acid. A Basic Local 
      Alignment Search Tool (BLAST) search demonstrated that CarA homologs were 
      distributed in soil microorganisms but not intestinal ones. In addition to CarA, 
      two CarA homologs were cloned and heterologously expressed, and their biochemical 
      properties were determined. Furthermore, a crystal structure of one homolog was 
      determined. Together with the biochemical analysis, the crystal structure and a 
      mutagenesis analysis of CarA revealed the mechanisms underlying their substrate 
      specificity and catalytic reaction. Our study suggests that CarA and its homologs 
      play a crucial role in the metabolism of C-glycosides in nature.
FAU - Kumano, Takuto
AU  - Kumano T
AUID- ORCID: 0000-0002-9240-4074
AD  - Graduate School of Life and Environmental Sciences, University of Tsukuba, 
      Tsukuba, Ibaraki 305-8572, Japan.
AD  - Microbiology Research Center for Sustainability, University of Tsukuba, Tsukuba, 
      Ibaraki 305-8572, Japan.
FAU - Hori, Sanae
AU  - Hori S
AD  - Graduate School of Life and Environmental Sciences, University of Tsukuba, 
      Tsukuba, Ibaraki 305-8572, Japan.
FAU - Watanabe, Satomi
AU  - Watanabe S
AD  - Graduate School of Life and Environmental Sciences, University of Tsukuba, 
      Tsukuba, Ibaraki 305-8572, Japan.
FAU - Terashita, Yuzu
AU  - Terashita Y
AD  - Graduate School of Life and Environmental Sciences, University of Tsukuba, 
      Tsukuba, Ibaraki 305-8572, Japan.
FAU - Yu, Hong Yang
AU  - Yu HY
AD  - Structural Biology Research Center, Institute of Materials Structure Science, 
      High Energy Accelerator Research Organization (KEK), Tsukuba, Ibaraki 305-0801, 
      Japan.
AD  - School of High Energy Accelerator Science, The Graduate University for Advanced 
      Studies (SOKENDAI), Tsukuba, Ibaraki 305-0801, Japan.
FAU - Hashimoto, Yoshiteru
AU  - Hashimoto Y
AD  - Graduate School of Life and Environmental Sciences, University of Tsukuba, 
      Tsukuba, Ibaraki 305-8572, Japan.
AD  - Microbiology Research Center for Sustainability, University of Tsukuba, Tsukuba, 
      Ibaraki 305-8572, Japan.
FAU - Senda, Toshiya
AU  - Senda T
AUID- ORCID: 0000-0002-5153-928X
AD  - Structural Biology Research Center, Institute of Materials Structure Science, 
      High Energy Accelerator Research Organization (KEK), Tsukuba, Ibaraki 305-0801, 
      Japan.
AD  - School of High Energy Accelerator Science, The Graduate University for Advanced 
      Studies (SOKENDAI), Tsukuba, Ibaraki 305-0801, Japan.
AD  - Faculty of Pure and Applied Sciences, University of Tsukuba, Tsukuba, Ibaraki 
      305-8572, Japan.
FAU - Senda, Miki
AU  - Senda M
AUID- ORCID: 0000-0003-3863-4503
AD  - Structural Biology Research Center, Institute of Materials Structure Science, 
      High Energy Accelerator Research Organization (KEK), Tsukuba, Ibaraki 305-0801, 
      Japan; miki.senda@kek.jp kobayashi.m.fe@u.tsukuba.ac.jp.
FAU - Kobayashi, Michihiko
AU  - Kobayashi M
AD  - Graduate School of Life and Environmental Sciences, University of Tsukuba, 
      Tsukuba, Ibaraki 305-8572, Japan; miki.senda@kek.jp 
      kobayashi.m.fe@u.tsukuba.ac.jp.
AD  - Microbiology Research Center for Sustainability, University of Tsukuba, Tsukuba, 
      Ibaraki 305-8572, Japan.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Proc Natl Acad Sci U S A
JT  - Proceedings of the National Academy of Sciences of the United States of America
JID - 7505876
RN  - 0 (C-glycoside)
RN  - 0 (Cardiac Glycosides)
RN  - 0 (Glycosides)
RN  - 146-14-5 (Flavin-Adenine Dinucleotide)
RN  - CID8Z8N95N (Carmine)
RN  - EC 1.- (Oxidoreductases)
SB  - IM
MH  - Cardiac Glycosides/metabolism
MH  - Carmine/metabolism
MH  - Catalysis
MH  - Flavin-Adenine Dinucleotide/*metabolism
MH  - Glycosides/*metabolism
MH  - Metabolic Networks and Pathways/physiology
MH  - Microbacterium/*metabolism
MH  - Mutagenesis/physiology
MH  - Oxidoreductases/metabolism
MH  - Substrate Specificity
PMC - PMC8501837
OTO - NOTNLM
OT  - C-glycoside
OT  - microorganism
OT  - oxidase
COIS- The authors declare no competing interest.
EDAT- 2021/09/30 06:00
MHDA- 2021/12/15 06:00
PMCR- 2022/03/28
CRDT- 2021/09/29 06:02
PHST- 2021/08/23 00:00 [accepted]
PHST- 2021/09/29 06:02 [entrez]
PHST- 2021/09/30 06:00 [pubmed]
PHST- 2021/12/15 06:00 [medline]
PHST- 2022/03/28 00:00 [pmc-release]
AID - 2106580118 [pii]
AID - 202106580 [pii]
AID - 10.1073/pnas.2106580118 [doi]
PST - ppublish
SO  - Proc Natl Acad Sci U S A. 2021 Oct 5;118(40):e2106580118. doi: 
      10.1073/pnas.2106580118.