DOI QR코드

DOI QR Code

Production and Application of Recombinant Agarase

재조합 한천 분해효소의 생산과 응용

Kim, Se Won;Hong, Chae-Hwan;Yun, Na Kyong;Shin, Hyun-Jae
김세원;홍채환;윤나경;신현재

  • Received : 2016.05.18
  • Accepted : 2016.06.03
  • Published : 2016.06.30

Abstract

The hydrolysis of biomass to fermentable sugar (saccharification) and to oligosaccharide is an essential process in biotechnology including biorefinery and biofood. Various macroalgae are commercially cultivated in several Asian countries as a useful resource for food and agar production. Agar is a major component of the cell walls of red algae that can be hydrolyzed by agarase. Agarases are classified into ${\alpha}$-agarase (E.C. 3.2.1.158) and ${\beta}$-agarase (E.C. 3.2.1.81) according to the cleavage pattern and grouped in the glycoside hydrolase (GH) family (GH-16, GH-58, GH-86, GH-96, and GH-118) based on the amino acid sequences of the proteins. Agarases have been isolated from various bacteria found in seawater and marine sediments. To increase productivity of the enzyme, a research on recombinant enzymes has been done. The application of recombinant agarase can be possible in the various filed such as energy, food, cosmetics, medical and so on. This paper reviews the source, biochemical characteristics and production system of recombinant agarases for further study.

Keywords

Red Algae;Agar;Recombinant Agarase;Saccharification;Oligosaccharides;GH family

References

  1. Araki C. 1959. Seaweed polysaccharides. In: Wolfrom ML (ed) Carbohydrate chemistry of substances of biological interest. Pergamon, London, pp 15-30.
  2. Araki, T., Hayakawa, M., Lu, Z., Karita, S. and Morishita, T. 1998. Purification and characterization of agarases from a marine bacterium, Vibrio sp. PO-303. J. Mar. Biotechnol. 6, 260-265.
  3. Chi, W. J., Chang, Y. K. and Hong, S. K. 2012. Agar degradation by microorganisms and agar-degrading enzymes. Appl. Microbiol. Biotechnol. 94, 917-930. https://doi.org/10.1007/s00253-012-4023-2
  4. Chi, W. J., Lee, C. R., Dugerjonjuu, S., Park, J. S., Kang, D. K. and Hong, S. K. 2015. Biochemical characterization of a novel iron-dependent GH16 ${\beta}$-agarase, AgaH92, from an agarolytic bacterium Pseudoalteromonas sp. H9. FEMS Microbiol. Lett. 362(7), fnv035.
  5. Chi, W. J., Park, da. Y., Seo, Y. B., Chang, Y. K., Lee, S. Y. and Hong, S. K. 2014. Cloning, expression, and biochemical characterization of a novel GH16 ${\beta}$-agarase AgaG1 from Alteromonas sp. GNUM-1. Appl. Microbiol. Biotechnol. 98(10), 4545-4555. https://doi.org/10.1007/s00253-014-5510-4
  6. Cregg, J. M., Cereghino, J. L., Shi, J. and Higgins, D. R. 2000. Recombinant Protein Expression in Pichia pastoris. Mol. Biotechnol. 6(1), 23-52.
  7. Cui, F., Dong, S, Shi, X., Zhao, X. and Zhang, X. H. 2014. Overexpression and characterization of a novel the rmostable ${\beta}$-agarase YM01-3, from marine bacterium Catenovulum agarivorans YM01(T). Mar. Drugs. 12(5), 2731-2747. https://doi.org/10.3390/md12052731
  8. Dong, J., Hashikawa, S., Konishi, T., Tamaru, Y. and Araki, T. 2006. Cloning of the novel gene encoding beta-agarase C from a marine bacterium, Vibrio sp. strain PO-303, and characterization of the gene product. Appl. Environ. Microbiol. 72(9), 6399-6401. https://doi.org/10.1128/AEM.00935-06
  9. Dong, J., Tamaru, Y. and Arakia T. 2007. A unique beta-agarase, AgaA, from a marine bacterium, Vibrio sp. strain PO-303. Appl. Microbiol. Biotechnol. 74(6), 1248-1255. https://doi.org/10.1007/s00253-006-0781-z
  10. Dong, J., Tamaru, Y. and Araki, T. 2007. Molecular cloning, expression, and characterization of a beta-agarasegene, agaD, from a marine bacterium, Vibrio sp. strain PO-303. Biosci. Biotechnol. Biochem. 71(1), 38-46. https://doi.org/10.1271/bbb.60304
  11. Duckworth, M. and Yaphe, W. 1971. The structure of agar: Part I. Fractionation of a complex mixture of polysaccharides. Carbohydr. Res. 16, 189-197. https://doi.org/10.1016/S0008-6215(00)86113-3
  12. Ekborg, N. A., Taylor, L. E., Longmire, A. G., Henrissat, B., Weiner, R. M. and Hutcheson, S.W. 2006. Genomic and proteomic analyses of the agarolytic system expressed by Saccharophagus degradans2-40. Appl. Environ. Microbiol. 72(5), 3396-3405. https://doi.org/10.1128/AEM.72.5.3396-3405.2006
  13. Fu, X. and Kim, S. 2010. Agarases: review of major sources, categories, purification method, enzyme characteristics and application. Mar. Drugs. 8, 200-218. https://doi.org/10.3390/md8010200
  14. Gosalbes, M. J., Esteban, C. D., Galan, J. L. and Perez-Martinez, G. 2000. Integrative food-grade expression system based on the lactose regulon of Lactobacillus casei. Appl. Environ. Microbiol. 66(11), 4822-4828. https://doi.org/10.1128/AEM.66.11.4822-4828.2000
  15. Hou, Y., Chen, X., Chan, Z and Zeng, R. 2015. Expression and characterization of a thermostable and pH-stable ${\beta}$-agarase encoded by a new gene from Flammeovirga pacifica WPAGA1. Process Biochem. 50, 1068-1075. https://doi.org/10.1016/j.procbio.2015.04.005
  16. Hsu, P. H., Wei, C. H., Lu, W. J., Shen, F., Pan, C.L. and Lin, H. T. 2015. Extracellular production of a novel endo-${\beta}$-agarase AgaA from Pseudomonas vesicularis MA103 that cleaves agarose into neoagarotetraose and neoagarohexaose. Int. J. Mol. Sci. 16(3), 5590-5603. https://doi.org/10.3390/ijms16035590
  17. Jam, M., Flament, D., Allouch, J., Potin, P., Thion, L., Kloareg, B., Czjzek, M., Helbert, W., Michel, G. and Barbeyron, T. 2005. The endo-beta-agarases AgaA and AgaB from the marine bacterium Zobellia galactanivorans: two paralogue enzymes with different molecular organizations and catalytic behaviours. Biochem. J. 385, 703-713. https://doi.org/10.1042/BJ20041044
  18. Jeon, M. J., Kim, A. R., Lee, D. G. and Lee, S. H. 2012. Cloning, Expression, and Characterization of a Novel GH-16 ${\beta}$-Agarase from Agarivorans sp. JA-1. Kor. J. Life Science. 22(11), 1545-1551. https://doi.org/10.5352/JLS.2012.22.11.1545
  19. Kim, H. T., Lee, S., Lee, D., Kim, H. S., Bang, W. G., Kim, K. H. and Choi, I. G. 2010. Overexpression and molecular characterization of Aga50D from Saccharophagus degradans2-40: an exo-type beta-agarase producing neoagarobiose. Appl. Microbiol. Biotechnol. 86(1), 227-34. https://doi.org/10.1007/s00253-009-2256-5
  20. Kobayashi, R., Takisada, M., Suzuki, T., Kirimura, K. and Usami, S. 1997. Neoagarobiose as a novel moisturizer with whitening effect. Biosci. Biotechnol. Biochem. 61, 162-163. https://doi.org/10.1271/bbb.61.162
  21. Lee, D. G., Jang, M. K., Lee, O. H., Kim, N. Y., Ju, S. A. and Lee, S. H. 2008. Over-production of a glycoside hydrolase family 50 ${\beta}$-agarase from Agarivorans sp. JA-1 in Bacillus subtilis and the whitening effect of its product. Biotechnol. Lett. 30, 911-918. https://doi.org/10.1007/s10529-008-9634-4
  22. Lee, D. G., Jeon, M. J. and Lee, S. H. 2012. Cloning, expression, and characterization of a glycoside hydrolase family118 beta-agarase from Agarivorans sp. JA-1. J. Microbiol. Biotechnol. 22(12), 1692-1697. https://doi.org/10.4014/jmb.1209.09033
  23. Lee, D. G., Park, G. T., Kim, N. Y., Lee, E. J., Jang, M. K., Shin, Y. G., Park, G. S., Kim, T. M., Lee, J. H., Lee, J. H., Kim, S. J. and Lee, S. H. 2006. Cloning, expression, and characterization of a glycoside hydrolase family 50 beta-agarase from a marine Agarivoransisolate. Biotechnol. Lett. 28(23), 1925-1932. https://doi.org/10.1007/s10529-006-9171-y
  24. Lee, Y., Oh, C., De, Zoysa M., Kim, H., Wickramaarachc hi, W. D., Whang, I., Kang, D. H., and Lee, J. 2013. Molecular cloning, overexpression, and enzymatic characterization of glycosyl hydrolase family16 ${\beta}$-Agarase from marine bacterium Saccharophagus sp. AG21 in Escherichia coli. J. Microbiol. Biotechnol. 23(7), 913-922. https://doi.org/10.4014/jmb.1209.09009
  25. Li, G., Sun, M., Wu, J., Ye, M., Ge, X., Wei, W., Li, H. and Hu, F. 2015. Identification and biochemical characterization of a novel endo-type ${\beta}$-agarase AgaW from Cohnella sp. strain L GH. Appl. Microbiol. Biotechnol. 99(23), 10019-10029. https://doi.org/10.1007/s00253-015-6869-6
  26. Li, J. and Sha, Y. 2015. Expression and enzymatic characterization of a cold-adapted ${\beta}$-agarase from Antarctic bacterium Pseudoalteromonas sp. NJ21. Chinese J. Oceanol. Limnol. 33(2), 319-327. https://doi.org/10.1007/s00343-015-4072-3
  27. Liao, L., Xu, X. W., Jiang, X. W., Cao, Y., Yi, N., Huo, Y. Y., Wu, Y. H., Zhu, X. F., Zhang, X. Q. and Wu, M. 2011. Cloning, expression, and characterization of a new beta-agarase from Vibrio sp. strain CN41. Appl. Environ. Microbiol. 77(19), 7077-7079. https://doi.org/10.1128/AEM.05364-11
  28. Lin, B., Lu, G., Zheng, Y., Xie, W., Li, S. and Hu, Z. 2012. Gene cloning, expression and characterization of a neoagarotetraose-producing ${\beta}$-agarase from the marine bacterium Agarivorans sp. HZ105. World J. Microbiol. Biotechnol. 28(4), 1691-1697. https://doi.org/10.1007/s11274-011-0977-y
  29. Liu, N., Mao, X., Yang, M., Mu, B. and Wei, D. 2014. Genecloning, expression and characterisation of a new ${\beta}$-agarase, AgWH50C, producing neoagarobiose from A garivorans gilvus WH0801. World J. Microbiol. Biotechnol. 30(6), 1691-1698. https://doi.org/10.1007/s11274-013-1591-y
  30. Long, M., Yu, Z. and Xu, X. 2010. A novel beta-agarase with high pH stability from marine Agarivorans sp. LQ48. Mar. Biotechnol. 12(1), 62-69. https://doi.org/10.1007/s10126-009-9200-7
  31. Lopes, T. S., Klootwijk, J., Veenstra, A. E., van der Aar, P. C., van Heerikhuizen, H., Raue, H. A. and Planta, R. J. 1989. High-copy-number integration into the ribosomal DNA of Saccharomyces cerevisiae: a new vector for high-level expression. Gene. 79(2), 199-206. https://doi.org/10.1016/0378-1119(89)90202-3
  32. Lu, X., Chu, Y., Wu, Q., Gu, Y., Han, F. and Yu, W. 2009. Cloning, expression and characterization of a new agarase-encoding gene from marine Pseudoalteromonas sp. Biotechnol Lett. 31(10), 1565-1570. https://doi.org/10.1007/s10529-009-0042-1
  33. Morrice, L. M., McLean, M. W., Long, W. F. and Williamson, F. B. 1983. Beta-agarases I and II from Pseudomonas atlantica. Substrate specificities. Eur. J. Biochem. 137(1-2), 149-154. https://doi.org/10.1111/j.1432-1033.1983.tb07808.x
  34. Oh, C., Nikapitiya, C., Lee, Y., Whang, I., Kim, S. J., Kang, D. H. and Lee, J. 2010. Cloning, purification and biochemical characterization of beta agarase from the marine bacterium Pseudoalteromonas sp. AG4. J. Ind. Microbiol. Biotechnol. 37(5), 483-494. https://doi.org/10.1007/s10295-010-0694-9
  35. Oh. C., Nikapitiya, C., Lee, Y., Whang, I., Kang, D. H., Heo, S. J., Choi, Y. U. and Lee, J. 2010. Molecular cloning, characterization and enzymatic properties of a novel beta-agarase from a marine isolate Psudoalteromonas SP. AG52. Braz. J. Microbiol. 41(4), 876-889. https://doi.org/10.1590/S1517-83822010000400006
  36. Ohta, Y., Hatada, Y., Miyazaki, M., Nogi, Y., Ito, S. and Horikoshi, K. 2005. Purification and characterization of a novel ${\alpha}$-agarase from a Thalassomonas sp. Curr. Microbiol. 50, 212-216. https://doi.org/10.1007/s00284-004-4435-z
  37. Potin, P., Richard, C., Rochas, C. and Kloareg, B. 1993. Purification and characterization of the ${\alpha}$-agarase from Alteromonas agarlyticus (Cataldi) comb. nov., strain GJ1B. Eur. J. Biochem. 214, 599-607. https://doi.org/10.1111/j.1432-1033.1993.tb17959.x
  38. Seo, Y. B., Lu, Y., Chi, W. J., Park, H. R., Jeong, K. J., Hong. S. K. and Chna. Y. K. 2014. Heterologous expression of a newly screened ${\beta}$-agarase from Alteromonas sp. GNUM1 in Escherichia coli and its application for agarose degradation. Process Biochem. 49, 430-436. https://doi.org/10.1016/j.procbio.2013.12.014
  39. Seok, J. H., Kim, H. S., Hatada, Y., Nam, S. W. and Kim, Y. H. 2012. Construction of an expression system for the secretory production of recombinant ${\alpha}$-agarase in yeast. Biotechnol. Lett. 34(6), 1041-1049. https://doi.org/10.1007/s10529-012-0864-0
  40. Seok, J. H., Park, H. G., Lee, S. H., Nam, S. W., Jeon, S. J., Kim, J. H. and Kim Y. H. 2010. High level secretory expression of recombinant ${\beta}$-agarase from Zobellia galactanivorans in Pichia pastoris. Kor. J. Microbiol. Biotechnol. 38(1), 40-45.
  41. Shi, Y. L., Lu, X. Z. and Yu W. G. A. 2008. A new ${\beta}$-agarase from marine bacterium Janthinobacterium sp. SY12. World J. Microbiol. Biotechnol. 24, 2659-2664. https://doi.org/10.1007/s11274-008-9792-5
  42. Tao, X., Jing, W., Kim, K. S., Yu, A. and Lee, Y. C. 2008. Cloning, expression and characterization of ${\beta}$-agarase gene from a marine bacterium, Pseudoalteromonas sp. JT-6. Kor. J. Life Science. 18(5), 625-630. https://doi.org/10.5352/JLS.2008.18.5.625
  43. Tawara, M., Sakatoku, A., Tiodjio, R. E., Tanaka, D. and Nakamura, S. 2015. Cloning and characterization of a novel agarase from a newly isolated bacterium Simiduia sp. strain TM-2 able to degrade various seaweeds. Appl. Biochem. Biotechnol. 177(3), 610-623. https://doi.org/10.1007/s12010-015-1765-1
  44. Temuujin, U., Chi, W. J., Chang, Y. K. and Hong, S. K. 2012. Identification and biochemical characterization of Sco3487 from Streptomyces coelicolor A3(2), an exoand endo-type ${\beta}$-agarase-producing neoagarobiose. J. Bacteriol. 194(1), 142-149. https://doi.org/10.1128/JB.05978-11
  45. Temuujin, U., Chi, W. J., Lee, S. Y., Chang, Y. K. and Hong, S. K.2011. Overexpression and biochemical chara cterization of DagA from Streptomyces coelicolor A3(2): an endo-type ${\beta}$-agarase producing neoagarotetraose and neoagarohexaose. Appl. Microbiol. Biotechnol. 92(4), 749-759. https://doi.org/10.1007/s00253-011-3347-7
  46. Xie, W., Lin, B., Zhou, Z., Lu, G., Lun, J., Xia, C., Li, S. and Hu, Z. 2013. Characterization of a novel ${\beta}$-agarase from an agar-degrading bacterium Catenovulum sp. X3. Appl. Microbiol. Biotechnol. 97(11), 4907-4915. https://doi.org/10.1007/s00253-012-4385-5
  47. Yang, J. I., Chen, L. C., Shih, Y. Y., Hsieh, C., Chen, C. Y., Chen, W. M. and Chen, C. C. 2011. Cloning and characterization of ${\beta}$-agarase AgaYT from Flammeovirgayae yamensisstrain YT. J. Bacteriol. 112(3), 225-232.
  48. Zhang, W. W. and Sun, L. 2007. Cloning, characterization, and molecular application of a beta-agarase gene from Vibrio sp. strain V134. Appl. Environ. Microbiol. 73(9), 2825-2831. https://doi.org/10.1128/AEM.02872-06

Acknowledgement

Supported by : (주)현대엔지비