Abstract
Our study investigated the biological functions of three neoagarooligosaccharides (NAOs), neoagarobiose (NA2), neoagarotetraose (NA4), and neoagarohexaose (NA6). NAOs were prepared by hydrolyzing agar with the recombinant β-agarases, DagA and DagB, from Streptomyces coelicolor A3(2). An in vitro DPPH (diphenyl-2,4,6-trinitrophenyliminoazanium) radical scavenging assay revealed that none of the NAOs had any significant antioxidative activity. We also assessed the antibacterial activity of each NAO against several gram-negative and gram-positive bacteria. This revealed weak effects against four of the seven tested strains. NAO treatment resulted in significant inhibition of α-glucosidase, with the strongest effect observed in a NA4 and NA6 mixture, and a decreasing effect was observed in the order NA2>NA4>NA6. Little inhibition was observed against α-amylase, with an effect significantly lower than that of acarbose. No observable cytotoxicity was found in immortalized Melan-a mouse melanocytes by NAO treatment up to a concentration of 2,000 μg/mL. However, melanin synthesis was significantly reduced by NAO treatment, with the strongest effect observed in a NA4 and NA6 mixture and decreasing effects observed in the order NA6>NA4>NA2. The tyrosinase activity of Melan-a cells was also consistently decreased by NAO treatment. Among the tested factors, treatment of NA4 and NA6 mixture showed the highest inhibition effects against α-glucosidase and tyrosinase, and melanin biosynthesis. Further in vivo study to investigate the role of NAO in these biological functions will be necessary for future biotechnological applications.
Similar content being viewed by others
References
Araki, C. H. (1937) Acetylation of agar like substance of Gelidium amansii. J. Chem. Soc. 58: 1338–1350.
Chi, W. J., Y. K. Chang, and S. K. Hong (2012) Agar degradation by microorganisms and agar-degrading enzymes. Appl. Microbiol. Biotechnol. 94: 917–930.
Giordano, A., G. Andreotti, A. Tramice, and A. Trincone (2006) Marine glycosyl hydrolases in the hydrolysis and synthesis of oligosaccharides. Biotechnol. J. 1: 511–530.
Potin, P., C. Richard, C. Rochas, and B. Kloareg (1993) Purification and characterization of the α-agarase from Alteromonas agarlyticus (Cataldi) comb. nov., strain GJ1B. Eur. J. Biochem. 214: 599–607.
Kirimura, K., N. Masuda, Y. Iwasaki, H. Nakagawa, R. Kobayashi, and S. Usami (1999) Purification and characterization of a novel β-agarase from an alkalophilic bacterium, Alteromonas sp. E-1. J. Biosci. Bioeng. 87: 436–441.
Temuujin, U., S. Y. Lee, Y. K. Chang, W. J. Chi, and S. K. Hong (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.
Temuujin, U., W. J. Chi, Y. K. Chang, and S. K. Hong (2012) Identification and biochemical characterization of Sco3487 from Streptomyces coelicolor A3(2), an exo-and endo-type β-agaraseproducing neoagarobiose. J. Bacteriol. 194: 142–149.
Kieser, H., M. J. Bibb, M. J. Buttner, F. K. Chater, and D. A. Hopwood (2000) Practical Streptomyces genetics. The John Innes Foundation, Norwich.
Sengupta, S., M. L. Jana, D. Sengupta, and A. K. Naskar (2000) A note on the estimation of microbial glycosidase activities by dinitrosalicylic acid reagent. Appl. Microbiol. Biotechnol. 53: 732–735.
Park, J., S. K. Hong, and Y. K. Chang (2014) Production of DagA, a β-agarase, by Streptomyces lividans in glucose medium or mixed-sugar medium simulating microalgae hydrolysate. J. Microbiol. Biotechnol. 24: 1622–1628.
Hossain, H., U. K. Karmakar, S. K. Biswas, A. F. Shahid-Ud-Daula, I. A. Jahan, T. Adnan, and A. Chowdhury (2013) Antinociceptive and antioxidant potential of the crude ethanol extract of the leaves of Ageratum conyzoides grown in Bangladesh. Pharm. Biol. 51: 893–898.
Xu, M.-L., L. Wang, G. F. Xu, and M.-H. Wang (2011) Antidiabetes and angiotensin converting enzyme inhibitory activity of Sonchus asper (L) Hill extract. Kor. J. Pharmacogn. 42: 61–67.
Bennett, D. C., P. J. Cooper, and I. R. Hart (1987) A line of nontumorigenic mouse melanocyte, syngeneic with the B16 melanoma and requiring a tumor promoter for growth. Int. J. Cancer. 39: 414–418.
Berridge, M. V., A. S. Tan, K. D. Mccoy, and R. Rui Wang (1996) The biochemical and cellular basis of cell proliferation assays that use tetrazolium salts. Biochemica 96: 14–19.
Busca, R., C. Bertolotto, J. P. Ortonne, and R. Ballotti (1996) Inhibition of the phosphatidylinositol 3-kinase/p70(S6)-kinase pathway induces B16 melanoma cell differentiation. J. Biol. Chem. 271: 31824–31830.
Jeon, Y. J. and S. K. Kim (2001) Potential immune-stimulating effect of antitumoral fraction of chitosan oligosaccharides. J. Chitin. Chitosan 6: 163–167.
Kim, M. H., H. Y. Kim, W. K. Kim, J. Y. Kim, and S. H. Kim (2001) Effects of soyoligosaccharides on blood glucose and lipid metabolism in streptozotocin-induced diabetic rats. Kor. Nut. Soc. 34: 3–13.
Enoki, T., T. Tominaga, F. Takashima, H. Ohnogi, H. Sagawa, and I. Kato (2012) Anti-tumor-promoting activities of agaro-oligosaccharides on two-stage mouse skin carcinogenesis. Biol. Pharm. Bull. 35: 1145–1149.
Chen, H., X. Yan, P. Zhu, and J. Lin (2006) Antioxidant activity and hepatoprotective potential of agaro-oligosaccharides in vitro and in vivo. Nutri. J. 5: 31.
Kobayashi, R., M. Takisada, T. Suzuki, K. Kirimura, and S. Usami (1997) Neoagarobiose as a novel moisturizer with whitening effect. Biosci. Biotechnol. Biochem. 61: 162–163.
Jang, M. K., D. G. Lee, N. Y. Kim, K. H. Yu, H. J. Jang, S. W. Lee, H. J. Jang, Y. J. Lee, and S. H. Lee (2009) Purification and characterization of neoagarotetraose from hydrolyzed agar. J. Microbiol. Biotechnol. 19: 1197–1200.
Hong, S. J., J. H. Lee, E. J. Kim, H. J. Yang, J. S. Park, and S. K. Hong (2017) Anti-obesity and anti-diabetic effect of neoagarooligosaccharides on high-fat diet-induced obesity in mice. Mar. Drugs 15(4). pii: E90. doi: 10.3390/md15040090
Hong, S. J., J. H. Lee, E. J. Kim, H. J. Yang, J. S. Park, and S. K. Hong (2017) Toxicological evaluation of neoagarooligosaccharides prepared by enzymatic hydrolysis of agar. Regul. Toxicol. Pharmacol. 90: 9–21.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Hong, S.J., Lee, JH., Kim, E.J. et al. In vitro and in vivo investigation for biological activities of neoagarooligosaccharides prepared by hydrolyzing agar with β-agarase. Biotechnol Bioproc E 22, 489–496 (2017). https://doi.org/10.1007/s12257-017-0049-8
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12257-017-0049-8