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Purification and Characterization of a Novel α-Agarase from a Thalassomonas sp.

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Abstract

An agar-degrading Thalassomonas bacterium, strain JAMB-A33, was isolated from the sediment off Noma Point, Japan, at a depth of 230 m. A novel α-agarase from the isolate was purified to homogeneity from cultures containing agar as a carbon source. The molecular mass of the purified enzyme, designated as agaraseA33, was 85 kDa on both SDS-PAGE and gel-filtration chromatography, suggesting that it is a monomer. The optimal pH and temperature for activity were about 8.5 and 45°C, respectively. The enzyme had a specific activity of 40.7 U/mg protein. The pattern of agarose hydrolysis showed that the enzyme is an endo-type α-agarase, and the final main product was agarotetraose. The enzyme degraded not only agarose but also agarohexaose, neoagarohexaose, and porphyran.

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Literature Cited

  1. J Allouch M Jam W Helbert T Barbeyron B Kloareg B Henrissat et al. (2003) ArticleTitleThe three-dimensional structures of two β-agarases J Biol Chem 278 47171–47180 Occurrence Handle10.1074/jbc.M308313200 Occurrence Handle1:CAS:528:DC%2BD3sXovFKktLY%3D Occurrence Handle12970344

    Article  CAS  PubMed  Google Scholar 

  2. M Duckworth W Yaphe (1971) ArticleTitleStructure of agar. I. Fractionation of a complex mixture of polysaccharides Carbohydr Res 1 189–197 Occurrence Handle10.1016/S0008-6215(00)86113-3

    Article  Google Scholar 

  3. JC Ha GT Kim SK Kim TK Oh JH Yu IS Kong (1997) ArticleTitleβ-agarase from Pseudomonas sp. W7: Purification of the recombinant enzyme from Escherichia coli and the effects of salt on its activity Biotechnol Appl Biochem 26 1–6 Occurrence Handle1:CAS:528:DyaK2sXlsVGkurY%3D Occurrence Handle9261996

    CAS  PubMed  Google Scholar 

  4. A Hosoda M Sakai S Kanazawa (2003) ArticleTitleIsolation and characterization of agar-degrading Paenibacillus spp. associated with the rhizosphere of spinach Biosci Biotechnol Biochem 67 1048–1055 Occurrence Handle10.1271/bbb.67.1048 Occurrence Handle1:CAS:528:DC%2BD3sXktlygtbs%3D Occurrence Handle12834282

    Article  CAS  PubMed  Google Scholar 

  5. I Kato (2000) ArticleTitleAntioxidative and antitumorigenic properties of agaro-oligosaccharide Bio Industry 17 13–19 Occurrence Handle1:CAS:528:DC%2BD3cXntFyktLc%3D

    CAS  Google Scholar 

  6. K Kendall J Cullum (1984) ArticleTitleCloning and expression of an extracellular-agarase from Streptomyces coelicolor A3(2) in Streptomyces lividans 66 Gene 29 315–321 Occurrence Handle10.1016/0378-1119(84)90060-X Occurrence Handle1:CAS:528:DyaL2MXhtlGj Occurrence Handle6489757

    Article  CAS  PubMed  Google Scholar 

  7. BJ Kim HJ Kim SH Ha SH Hwang DS Byun TH Lee et al. (1999) ArticleTitlePurification and characterization of β-agarase from marine bacterium Bacillus cereus ASK202 Biotechnol Lett 21 1011–1015 Occurrence Handle10.1023/A:1005604618242 Occurrence Handle1:CAS:528:DyaK1MXotFSitbs%3D

    Article  CAS  Google Scholar 

  8. YH Kim YS Kim JR Lee EK Lee JH Seu (1993) ArticleTitleEnzymatic characteristics of an extracellular agarase of Cytophaga sp. KY-1 and molecular cloning of the agarase gene J Microbiol Biotechnol 3 31–38 Occurrence Handle1:CAS:528:DyaK2cXmslCqtrg%3D

    CAS  Google Scholar 

  9. UK Laemmli (1970) ArticleTitleCleavage of structural proteins during the assembly of the head of bacteriophage T4 Nature 227 680–685 Occurrence Handle1:CAS:528:DC%2BD3MXlsFags7s%3D Occurrence Handle5432063

    CAS  PubMed  Google Scholar 

  10. LM Morrice MW McLean WF Long FB Williamson (1983) ArticleTitleβ-agarases I and II from Pseudomonas atlantica. Substrate specificities Eur J Biochem 137 149–154 Occurrence Handle10.1111/j.1432-1033.1983.tb07808.x Occurrence Handle1:CAS:528:DyaL2cXmtl2qug%3D%3D Occurrence Handle6653550

    Article  CAS  PubMed  Google Scholar 

  11. T Naganuma DA Coury M Poline-Fuller A Gibor K Horikoshi (1993) ArticleTitleCharacterization of agarolytic Microscilla isolates and their extracellular agarases Syst Appl Microbiol 16 183–190 Occurrence Handle1:CAS:528:DyaK2cXhsFyhtLk%3D

    CAS  Google Scholar 

  12. Y Ohta Y Hatada Y Nogi Z Li HM Zhang S Ito et al. (2004a) ArticleTitleThermostable β-agarase from a deep-sea Microbulbifer isolate J Appl Glycosci 51 203–210

    Google Scholar 

  13. Y Ohta Y Hatada Y Nogi M Miyazaki Z Li M Akita et al. (2004b) ArticleTitleEnzymatic properties and nucleotide and amino acid sequences of a thermostable β-agarase from a novel species of deep-sea Microbulbifer Appl Microbiol Biotechnol 64 505–514 Occurrence Handle10.1007/s00253-004-1573-y Occurrence Handle1:CAS:528:DC%2BD2cXjtFyis7o%3D

    Article  CAS  Google Scholar 

  14. P Potin C Richard C Rochas B Kloareg (1993) ArticleTitlePurification and characterization of the α-agarase from Alteromonas agarlyticus (Cataldi) comb nov., strain GJ1B. Eur J Biochem 214 599–607

    Google Scholar 

  15. C Rochas P Potin B Kloareg (1994) ArticleTitleNMR spectroscopic investigation of agarose oligomers produced by an α-agarase Carbohydr Res 253 69–77 Occurrence Handle10.1016/0008-6215(94)80056-1 Occurrence Handle1:CAS:528:DyaK2cXivF2isLs%3D Occurrence Handle8156559

    Article  CAS  PubMed  Google Scholar 

  16. DC Schroeder MA Jaffer VE Coyne (2003) ArticleTitleInvestigation of the role of a β (1–4) agarase produced by Pseudoalteromonas gracilis B9 in eliciting disease symptoms in the red alga Gracilaria gracilis Microbiology 149 2919–2929 Occurrence Handle10.1099/mic.0.26513-0 Occurrence Handle1:CAS:528:DC%2BD3sXosVGqurk%3D Occurrence Handle14523124

    Article  CAS  PubMed  Google Scholar 

  17. Y Sugano H Kodama I Terada Y Yamazaki M Noma (1994) ArticleTitlePurification and characterization of a novel enzyme, α-neoagarooligosaccharide hydrolase (α-NAOS hydrolase), from a marine bacterium, Vibrio sp. strain JT0107 J Bacteriol 176 6812–6818 Occurrence Handle1:STN:280:ByqD2M%2FisVM%3D Occurrence Handle7961439

    CAS  PubMed  Google Scholar 

  18. Y Sugano I Terada M Noma T Matsumoto (1993) ArticleTitlePurification and characterization of a new agarase from a marine bacterium, Vibrio sp. strain JT0107 Appl Environ Microbiol 59 1549– 1554

    Google Scholar 

  19. K Takahashi Y Hirano S Araki M Hattori (2000) ArticleTitleEmulsifying ability of porphyran prepared from dried nori, Porphyra yezoensis, a red alga J Agric Food Chem 48 2721–2725 Occurrence Handle10.1021/jf990990b Occurrence Handle1:CAS:528:DC%2BD3cXjvVOls7w%3D Occurrence Handle10898612

    Article  CAS  PubMed  Google Scholar 

  20. WG Weisburg SM Barns DA Pelletier DJ Lane (1991) ArticleTitle16S ribosomal DNA amplification for phylogenetic study J Bacteriol 173 697–703 Occurrence Handle1987160

    PubMed  Google Scholar 

  21. WP Winter J Yodh (1983) ArticleTitleInteraction of human hemoglobin and its variants with agar Science 221 175–178 Occurrence Handle1:CAS:528:DyaL3sXks1OjtLo%3D Occurrence Handle6190229

    CAS  PubMed  Google Scholar 

  22. Y Yoshizawa A Ametani J Tsunehiro K Nomura M Itoh F Fukui et al. (1995) ArticleTitleMacrophage stimulation activity of the polysaccharide fraction from a marine alga (Porphyra yezoensis): structure–function relationships and improved solubility Biosci Biotechnol Biochem 59 1933–1937 Occurrence Handle1:CAS:528:DyaK2MXptVKitrY%3D Occurrence Handle8534986

    CAS  PubMed  Google Scholar 

  23. Q Zhang N Li X Liu Z Zhao Z Li Z Xu (2004) ArticleTitleThe structure of a sulfated galactan from Porphyra haitanensis and its in vivo antioxidant activity Carbohydr Res 339 105–111 Occurrence Handle10.1016/j.carres.2003.09.015 Occurrence Handle1:CAS:528:DC%2BD3sXpsVKrs7o%3D Occurrence Handle14659676

    Article  CAS  PubMed  Google Scholar 

  24. Z Zhong A Toukdarian D Helinski V Knauf S Sykes JE Wilkinson et al. (2001) ArticleTitleSequence analysis of a 101-kilobase plasmid required for agar degradation by a Microscilla isolate Appl Environ Microbiol 67 5771–5779

    Google Scholar 

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Acknowledgments

We are grateful to Dr. Y. Sakano of Tokyo University of Agriculture and Technology for stimulating discussions.

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Authors and Affiliations

  1. Japan Agency for Marine-Earth Science and Technology (JAMSTEC), 2-15 Natsushima, Yokosuka , 237-0061, Japan

    Yukari Ohta, Yuji Hatada, Masayuki Miyazaki, Yuichi Nogi, Susumu Ito & Koki Horikoshi

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  1. Yukari Ohta
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  2. Yuji Hatada
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  3. Masayuki Miyazaki
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  4. Yuichi Nogi
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  5. Susumu Ito
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  6. Koki Horikoshi
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Correspondence to Yuji Hatada.

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Ohta, Y., Hatada, Y., Miyazaki, M. et al. Purification and Characterization of a Novel α-Agarase from a Thalassomonas sp.. Curr Microbiol 50, 212–216 (2005). https://doi.org/10.1007/s00284-004-4435-z

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  • DOI: https://doi.org/10.1007/s00284-004-4435-z

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