Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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NADP+-Dependent Dehydrogenase SCO3486 and Cycloisomerase SCO3480: Key Enzymes for 3,6-Anhydro-ʟ-Galactose Catabolism in Streptomyces coelicolor A3(2)

  • Tsevelkhorloo, Maral (Department of Biosciences and Bioinformatics, Myongji University) ;
  • Kim, Sang Hoon (Department of Animal Resources Science, Dankook University) ;
  • Kang, Dae-Kyung (Department of Animal Resources Science, Dankook University) ;
  • Lee, Chang-Ro (Department of Biosciences and Bioinformatics, Myongji University) ;
  • Hong, Soon-Kwang (Department of Biosciences and Bioinformatics, Myongji University)
  • Received : 2021.03.17
  • Accepted : 2021.04.05
  • Published : 2021.05.28
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Abstract

Agarose is a linear polysaccharide composed of ᴅ-galactose and 3,6-anhydro-ʟ-galactose (AHG). It is a major component of the red algal cell wall and is gaining attention as an abundant marine biomass. However, the inability to ferment AHG is considered an obstacle in the large-scale use of agarose and could be addressed by understanding AHG catabolism in agarolytic microorganisms. Since AHG catabolism was uniquely confirmed in Vibrio sp. EJY3, a gram-negative marine bacterial species, we investigated AHG metabolism in Streptomyces coelicolor A3(2), an agarolytic gram-positive soil bacterium. Based on genomic data, the SCO3486 protein (492 amino acids) and the SCO3480 protein (361 amino acids) of S. coelicolor A3(2) showed identity with H2IFE7.1 (40% identity) encoding AHG dehydrogenase and H2IFX0.1 (42% identity) encoding 3,6-anhydro-ʟ-galactonate cycloisomerase, respectively, which are involved in the initial catabolism of AHG in Vibrio sp. EJY3. Thin layer chromatography and mass spectrometry of the bioconversion products catalyzed by recombinant SCO3486 and SCO3480 proteins, revealed that SCO3486 is an AHG dehydrogenase that oxidizes AHG to 3,6-anhydro-ʟ-galactonate, and SCO3480 is a 3,6-anhydro-ʟ-galactonate cycloisomerase that converts 3,6-anhydro-ʟ-galactonate to 2-keto-3-deoxygalactonate. SCO3486 showed maximum activity at pH 6.0 at 50℃, increased activity in the presence of iron ions, and activity against various aldehyde substrates, which is quite distinct from AHG-specific H2IFE7.1 in Vibrio sp. EJY3. Therefore, the catabolic pathway of AHG seems to be similar in most agar-degrading microorganisms, but the enzymes involved appear to be very diverse.

Keywords

Acknowledgement

This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT & Future Planning (NRF-2020R1F1A1060789).

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