Article Search
닫기

Microbiology and Biotechnology Letters

보문(Article)

View PDF

Fermentation Microbiology (FM)  |  Fermentation Technology

Microbiol. Biotechnol. Lett. 2021; 49(3): 329-336

https://doi.org/10.48022/mbl.2105.05009

Received: May 12, 2021; Revised: July 5, 2021; Accepted: July 7, 2021

해양미생물 Cellulophga sp. J9-3이 생산하는 베타-아가레이즈의 분리 및 생화학적 특

Purification and Biochemical Characterization of β-agarase Produced by Marine Microorganism Cellulophga sp. J9-3

Da Som Kim1, Jong-Hee Kim2 and Won-Jae Chi1*

1Microorganism Resources Division, National Institute of Biological Resource, Incheon 22689, Republic of Korea 2Department of Food and Nutrition, Seoil University, Seoul 02192, Republic of Korea

Correspondence to :
Won-Jae Chi,     wjchi76@korea.kr

Cellulophga sp. J9-3, is a gram-negative, aerobic marine bacterium belonging to the family Flavobacteriaceae. In addition to cellulose degradability, the J9-3 strain is also capable of hydrolyzing agar in the solid and liquid medium, and the production of agarase in the presence of agarose can be remarkably induced by the bacterium. From the cell culture broth of Cellulophga sp. J9-3, ammonium sulfate precipitation and three kinds of column chromatography were successively performed to purify a specific agarase protein, the AgaJ93. Purified AgaJ93 showed the strongest hydrolyzing activity towards agarose (approximately 22%), and even displayed activity towards starch. AgaJ93 hydrolyzed agarose into neoagarotetraose and neoagarohexaose via various oligosaccharide intermediates, indicating that AgaJ93 is an endo-type β-agarase. AgaJ93 showed maximum activity at a pH of 7.0 and temperature of 35 °C. Its activity increased by more than six times in the presence of Co2+ ions. The N-terminal sequence of AgaJ93 showed 82% homology with the heat-resistant endo-type β-agarase Aga2 of Cellulophaga sp. W5C. However, the biochemical properties of the two enzymes were different. Therefore, AgaJ93 is expected to be a novel agarose, different from the previously reported β-agarases.

Keywords: Cellulophga sp. J9-3, neoagarotetraose, neoagarohexaose, agarose, agar

Graphical Abstract


  1. Knutsen SH, Myslabodski DE, Larsen B, Usov AI. 1994. A modified system of nomenclature for red algal galactans. Botanica Marina 37: 163-170.
  2. Chi WJ, Chang YK, Hong SK. 2012. Agar degradation by microorganisms and agar-degrading enzymes. Appl. Microbiol. Biotechnol. 94: 917-930.
    Pubmed
  3. Duckworth M, Yaphe W. 1971. Structure of agar: Part I. Fractionation of a complex mixture of polysaccharides. Carbohydr. Res. 16: 189-197.
  4. Hehemann JH, Correc G, Barbeyron T, Helbert W, Czjzek M, Michel G. 2010. Transfer of carbohydrate-active enzymes from marine bacteria to Japanese gut microbiota. Nature 464: 908912.
    Pubmed
  5. Park SH, Lee CR, Hong SK. 2020. Implications of agar and agarase in industrial applications of sustainable marine biomass. Appl. Microbiol. Biotechnol. 104: 2815-2832.
    Pubmed
  6. Wang W, Liu P, Hao C, Wu L, Wan W, Mao X. 2017. Neoagaro-oligosaccharide monomers inhibit inflammation in LPS-stimulated macrophages through suppression of MAPK and NF-κB pathways. Sci. Rep. 7: 44252.
    Pubmed KoreaMed
  7. Kang DR, Yoon GY, Cho J, Lee SJ, Lee SJ, Park HJ, et al. 2017. Neoagarooligosaccharides prevent septic shock by modulating A20-and cyclooxygenase-2-mediated interleukin-10 secretion in a septic-shock mouse model. Biochem. Biophys Res. Commun. 486: 998-1004.
    Pubmed
  8. Hong SJ, Lee JH, Kim EJ, Yang HJ, Park JS, Hong SK. 2017. Antiobesity and anti-diabetic effect of neoagarooligosaccharides on high-fat diet-induced obesity in mice. Mar. Drugs 15: 90102.
    Pubmed KoreaMed
  9. Lee MH, Jang JH, Yoon GY, Lee SJ, Lee MG, Kang TH, et al. 2017. Neoagarohexaose-mediated activation of dendritic cells via Toll-like receptor 4 leads to stimulation of natural killer cells and enhancement of antitumor immunity. BMB Rep. 50: 263268.
    Pubmed KoreaMed
  10. Yun EJ, Lee S, Kim JH, Kim BB, Kim HT, Lee SH, et al. 2013. Enzymatic production of 3,6-anhydro-L-galactose from agarose and its purification and in vitro skin whitening and anti-inflammatory activities. Appl. Microbiol. Biotechnol. 97: 2961-2970.
    Pubmed
  11. Yun EJ, Yu S, Kim KH. 2017. Current knowledge on agarolytic enzymes and the industrial potential of agar-derived sugars. Appl. Microbiol. Biotechnol. 101: 5581-5589.
    Pubmed
  12. Minegishi H, Shimane Y, Echigo A, Ohta Y, Hatada Y, Kamekura M, et al. 2013. Thermophilic and halophilic β-agarase from a halophilic archaeon Halococcus sp. 197A. Extremophiles 17:931-939.
    Pubmed KoreaMed
  13. Lee YR, Jung S, Chi WJ, Bae CH, Jeong BC, Hong SK, et al. 2018. Biochemical characterization of a novel GH86 β-agarase producing neoagarohexaose from Gayadomonas joobiniege G7. J. Microbiol. Biotechnol. 28: 284-292.
    Pubmed
  14. Han Z, Zhang Y, Yang J. 2019. Biochemical characterization of a new β-agarase from Cellulophaga algicola. Int. J. Mol. Sci. 20:2143-2157.
    Pubmed KoreaMed
  15. Jung S, Jeong BC, Hong SK, Lee CR. 2017. Cloning, expression, and biochemical characterization of a novel acidic GH16 βagarase, AgaJ11, from Gayadomonas joobiniege G7. Appl. Biochem. Biotechnol. 181: 961-971.
    Pubmed
  16. Chen X, Lin H, Jin M, Zeng R, Lin M. 2019. Characterization of a novel alkaline β-agarase and its hydrolysates of agar. Food Chem. 295: 311-319.
    Pubmed
  17. Kim DS, Chi W-J, Hong S-K. 2019. Molecular characterization of an endo-β-1,4-glucanase, CelAJ93, from the recently isolated marine bacterium, Cellulophaga sp. J9-3. Appl. Sci. 9: 40614073.
  18. Miller GL. 1959. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal. Chem. 31: 426-428.
  19. Laemmli UK. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680685.
    Pubmed
  20. Temuujin U, Chi WJ, Lee SY, Chang YK, Hong SK. 2011. Overexpression and biochemical characterization of DagA from Streptomyces coelicolor A3(2): an endo-type β-agarase producing neoagarotetraose and neoagarohexaose. Appl. Microbiol. Biotechnol. 92: 749-759.
    Pubmed
  21. Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang A, Miller W, et al. 1997. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res. 25:3389-3402.
    Pubmed KoreaMed
  22. Ramos KRM, Valdehuesa KNG, Nisola GM, Lee WK, Chung WJ. 2018. Identification and characterization of a thermostable endolytic β-agarase Aga2 from a newly isolated marine agarolytic bacteria Cellulophaga omnivescoria W5C. N. Biotechnol. 40:261-267.
    Pubmed
  23. Jung S, Lee CR, Chi WJ, Bae CH, Hong SK. 2017. Biochemical characterization of a novel cold-adapted GH39 β-agarase, AgaJ9, from an agar-degrading marine bacterium Gayadomonas joobiniege G7. Appl. Microbiol. Biotechnol. 101: 1965-1974.
    Pubmed
  24. Ramos KRM, Valdehuesa KNG, Bañares AB, Nisola GM, Lee WK, Chung WJ. 2020. Overexpression and characterization of a novel GH16 β-agarase (Aga1) from Cellulophaga omnivescoria W5C. Biotechnol. Lett. 42: 2231-2238.
    Pubmed

Starts of Metrics

Share this article on :

  • mail

Related articles in MBL

Most KeyWord ?

What is Most Keyword?

  • It is most registrated keyword in articles at this journal during for 2 years.