Skip to main content
SpringerLink
Log in

Characterization of a novel β-agarase from an agar-degrading bacterium Catenovulum sp. X3

  • Biotechnologically relevant enzymes and proteins
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

An agar-degrading bacterium, Catenovulum sp. X3, was isolated from the seawater of Shantou, China. A novel β-agarase gene agaXa was cloned from the strain Catenovulum sp. X3. The gene agaXa consists of 1,590 bp and encodes a protein of 529 amino acids, with only 40 % amino acid sequence identity with known agarases. AgaXa should belong to the glycoside hydrolase family GH118 based on the amino acid sequence similarity. The molecular mass of the recombinant AgaXa (rAgaXa) was estimated to be 52 kDa by sodium dodecyl sulfate–polyacrylamide gel electrophoresis. It had a maximal agarase activity at 52 °C and pH 7.4 and was stable over pH 5.0 ~ 9.0 and at temperatures below 42 °C. The K m and V max for agarose were 10.5 mg/ml and 588.2 U/mg, respectively. The purified rAgaXa showed endolytic activity on agarose degradation, yielding neoagarohexaose, neoagarooctaose, neoagarodecaose, and neoagarododecaose as the end products. The results showed that AgaXa has potential applications in agar degradation for the production of oligosaccharides with various bioactivities.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Bannikova GE, Lopatin SA, Varlamov VP, Kuznetsov BB, Kozina IV, Miroshnichenko ML, Chernykh NA, Turova TP, Bonch-Osmolovskaya EA (2008) The thermophilic bacteria hydrolyzing agar: characterization of thermostable agarase. Appl Biochem Microbiol 45:366–371

    Article  Google Scholar 

  • Cantarel BL, Coutinho PM, Rancurel C, Bernard T, Lombard V, Henrissat B (2009) The Carbohydrate-Active EnZyme database (CAZy): an expert resource for glycogenomics. Nucleic Acids Res 37:D233–238

    Article  CAS  Google Scholar 

  • Chen LCM, Craigiel JS, Xie ZK (1994) Protoplast production from Porphyra linearis using a simplified agarase procedure capable of commercial application. J Appl Phycol 6:35–39

    Article  CAS  Google Scholar 

  • Chi WJ, Chang YK, Hong SK (2012) Agar degradation by microorganisms and agar-degrading enzymes. Appl Microbiol Biotechnol 94:917–930

    Article  CAS  Google Scholar 

  • Dipakkore S, Reddy CRK, Jha B (2005) Production and seeding of protoplasts of Porphyra okhaensis (Bangiales, Rhodophyta) in laboratory culture. J Appl Phycol 17:331–337

    Article  CAS  Google Scholar 

  • Dong J, Hashikawa S, Konishi T, Tamaru Y, 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 72:6399–6401

    Article  CAS  Google Scholar 

  • Duckworth M, Yaphe W (1971) Structure of agar: part I. Fractionation of a complex mixture of polysaccharides. Carbohydr Res 16:189–197

    Article  CAS  Google Scholar 

  • Giordano A, Andreotti G, Tramice A, Trincone A (2006) Marine glycosyl hydrolases in the hydrolysis and synthesis of oligosaccharides. J Biotechnol 1:511–530

    Article  CAS  Google Scholar 

  • Finkelstein M, Rownd RH (1978) A rapid method for extracting DNA from agarose gels. Plasmid 1:557–562

    Article  CAS  Google Scholar 

  • Kim OS, Cho YJ, Lee K, Yoon SH, Kim M, Na H, Park SC, Jeon YS, Lee JH, Yi H, Won S, Chun J (2012) Introducing EzTaxon-e: a prokaryotic 16S rRNA gene sequence database with phylotypes that represent uncultured species. Int J Syst Evol Microbiol 62:716–721

    Article  CAS  Google Scholar 

  • Kobayashi R, Takimasa M, Suzuki T, Kirimura K, Usami S (1997) Neoagarobiose as a novel moisturizer with whitening effect. Biosci Biotechnol Biochem 61:162–163

    Article  CAS  Google Scholar 

  • Lane DJ (1991) 16S/23S rRNA sequencing. In: Stackebrandt E, Goodfellow M (eds) Nucleic acid techniques in bacterial systematics. John Wiley & Sons, Ltd., Chichester, pp 115–175

    Google Scholar 

  • Lee DG, Jang MK, Lee OH, Kim NY, Ju SA, Lee SH (2008) Overproduction 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

    Article  CAS  Google Scholar 

  • Lin B, Lu G, Zheng Y, Xie W, Li S, Hu Z (2012a) Aquimarina agarilytica sp. nov., a novel agarolytic species isolated from red alga. Int J Syst Evol Microbiol 62:869–873

    Article  CAS  Google Scholar 

  • Lin B, Lu G, Zheng Y, Xie W, Li S, Hu Z (2012b) Gene cloning, expression and characterization of a neoagarotetraose-producing β-agarase from a marine bacterium Agarivorans sp. HZ105. World J Microb Biot 28:1691–1697

    Article  CAS  Google Scholar 

  • Ma CP, Lu XZ, Shi C, Li J, Gu Y, Ma Y, Chu Y, Han F, Gong Q, Yu W (2007) Molecular cloning and characterization of a novel β-agarase, AgaB, from marine Pseudoalteromonas sp. CY24. J Biol Chem 282:3747–3754

    Article  CAS  Google Scholar 

  • Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31:426–428

    Article  CAS  Google Scholar 

  • Ohta Y, Hatada Y, Nogi Y, Miyazaki M, Li Z, Akita M, Hidaka Y, Goda S, Ito S, Horikoshi K (2004a) Enzymatic 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

    Article  CAS  Google Scholar 

  • Ohta Y, Nogi Y, Miyazaki M, Li Z, Hatada Y, Ito S, Horikoshi K (2004b) Enzymatic properties and nucleotide and amino acid sequences of a thermostable β-agarase from the novel marine isolate JAMB-A94. Biosci Biotechnol Biochem 68:1073–1081

    Article  CAS  Google Scholar 

  • Oppenheimer CH, ZoBell CE (1952) The growth and viability of sixty-three species of marine bacteria as influenced by hydrostatic pressure. J Mar Res 11:10–18

    Google Scholar 

  • Petersen TN, Brunak S, von Heijne G, Nielsen H (2011) SignalP 4.0: discriminating signal peptides from transmembrane regions. Nat Methods 8:785–786

    Article  CAS  Google Scholar 

  • Ren A, Xia ZX, Yu W, Zhou J (2010) Expression, crystallization and preliminary X-ray analysis of an anomeric inverting agarase from Pseudoalteromonas sp. CY24. Acta Cryst F66:1635–1639

    Google Scholar 

  • Wu SC, Wen TN, Pan CL (2005) Algal–oligosaccharide–lysates prepared by two bacterial agarases stepwise hydrolyzed and their antioxidative properties. Fish Sci 71:1149–1159

    Article  CAS  Google Scholar 

  • Yan S, Yu M, Wang Y, Shen C, Zhang XH (2011) Catenovulum agarivorans gen. nov., sp. nov., a peritrichously flagellated, chain-forming, agar-hydrolysing gammaproteobacterium from seawater. Int J Syst Evol Microbiol 61:2866–2873

    Article  CAS  Google Scholar 

  • Zhang WW, Sun L (2007) Cloning, characterization, and molecular application of a beta-agarase gene from Vibrio sp. strain V134. Appl Environ Microbiol 73:2825–2831

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by the National Natural Science Foundation of China (nos. 41076106 and 31200077), Guangdong Natural Science Foundation (no. S2011030005257), and the Key Science and Technology Innovation Project for University by the Department of Education of Guangdong Province (no. CXZD1124) and the Science & Technology Project of Guangdong Province (no. 2012A031100009).

Author information

Authors and Affiliations

  1. Department of Biology, Shantou University, Shantou, 515063, Guangdong, People’s Republic of China

    Wei Xie, Bokun Lin, Zhengrong Zhou, Guoyong Lu, Jingsheng Lun, Changyan Xia, Shengkang Li & Zhong Hu

Authors
  1. Wei Xie
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Bokun Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Zhengrong Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Guoyong Lu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jingsheng Lun
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Changyan Xia
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Shengkang Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Zhong Hu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Zhong Hu.

Additional information

Wei Xie, Bokun Lin, and Zhengrong Zhou contributed equally to this work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Xie, W., Lin, B., Zhou, Z. et al. Characterization of a novel β-agarase from an agar-degrading bacterium Catenovulum sp. X3. Appl Microbiol Biotechnol 97, 4907–4915 (2013). https://doi.org/10.1007/s00253-012-4385-5

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00253-012-4385-5

Keywords

Search

Navigation