Dual agarolytic pathways in a marine bacterium, Vibrio sp. strain EJY3: Molecular and enzymatic verification

Sora Yu, Eun Ju Yun, Dong Hyun Kim, So Young Park, Kyoung Heon Kim

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17 Citations (Scopus)


Vibrio sp. strain EJY3 is an agarolytic marine bacterium that catabolizes 3,6-anhydro-L-galactose (AHG), a monomeric sugar unit of agarose. While the AHG catabolic pathway in EJY3 has been discovered recently, the complete agarolytic system of EJY3 remains unclear. We have identified five enzymes, namely, the β-agarases VejGH50A, VejGH50B, VejGH50C, and VejGH50D and the α-neoagarooligosaccharide (NAOS) hydrolase VejGH117, involved in the agarolytic system of EJY3. Based on the characterization of recombinant enzymes and intracellular metabolite analysis, we found that EJY3 catabolizes agarose via two different agarolytic pathways. Among the four β-agarases of EJY3, VejGH50A, VejGH50B, and VejGH50C were found to be extracellular agarases, producing mainly neoagarotetraose (NeoDP4) and neoagarobiose. By detecting intracellular NeoDP4 in EJY3 grown on agarose, NeoDP4 was observed being taken up by cells. Intriguingly, intracellular NeoDP4 acted as a branching point for the two different downstream agarolytic pathways. First, via the well-known agarolytic pathway, NeoDP4 was depolymerized into monomeric sugars by the exo-type β-agarase VejGH50D and the α-NAOS hydrolase VejGH117. Second, via the newly found alternative agarolytic pathway, NeoDP4 was depolymerized into AHG and agarotriose (AgaDP3) by VejGH117, and AgaDP3 then was completely depolymerized into monomeric sugars by sequential reactions of the agarolytic β-galactosidases (ABG) VejABG and VejGH117. Therefore, by experimentally verifying agarolytic enzymatic activity and transport of NeoDP4 into EJY3 cells, we revealed that EJY3 possesses both the known pathway and the newly discovered alternative pathway that involves α-NAOS hydrolase and ABG. IMPORTANCE Agarose is the main polysaccharide of red macroalgae and is composed of galactose and 3,6-anhydro-L-galactose. Many marine bacteria possess enzymes capable of depolymerizing agarose into oligomers and then depolymerizing the oligomers into monomers. Here, we experimentally verified that both a wellknown agarolytic pathway and a novel agarolytic pathway exist in a marine bacterium, Vibrio sp. strain EJY3. In agarolytic pathways, agarose is depolymerized mainly into 4-sugar-unit oligomers by extracellular enzymes, which are then transported into cells. The imported oligomers are intracellularly depolymerized into galactose and 3,6-anhydro-L-galactose by two different agarolytic pathways, using different combinations of intracellular enzymes. These results elucidate the depolymerization routes of red macroalgal biomass in the ocean by marine bacteria and provide clues for developing industrial processes for efficiently producing sugars from red macroalgae.

Original languageEnglish
Article numbere02724-19
JournalApplied and environmental microbiology
Issue number1
Publication statusPublished - 2020 Mar 1

Bibliographical note

Funding Information:
We acknowledge grant support from the C1 Gas Refinery Program through the National Research Foundation of Korea (2016M3D3A1A01913268). The Research Fellow Grant funded by the National Research Foundation of Korea (2017R1A6A3A11035069) and the grants funded by Korea University are also acknowledged. This work was performed at the Korea University Food Safety Hall for the Institute of Biomedical Science and Food Safety.

Publisher Copyright:
© 2020 American Society for Microbiology. All Rights Reserved.


  • Agarolytic pathway
  • Agarolytic β-galactosidase
  • Agarose
  • Neoagarotetraose
  • Red macroalgae
  • Vibrio sp. strain ejy3
  • α-Neoagarooligosaccharide hydrolase

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Food Science
  • Biotechnology
  • Ecology


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