Skip to main content
SpringerLink
Log in

Cloning and Sequencing Analysis of Alginate Lyase Genes from the Marine Bacterium Vibrio sp. O2

  • Original Article
  • Published:
Marine Biotechnology Aims and scope Submit manuscript

Abstract

We isolated a new marine bacteria, which displayed alginate-depolymerizing activity in plate assays, from seawater in Mihonoseki Harbor, Japan. Analysis of the 16S ribosomal RNA gene sequence of one of the isolates proved that this alginate-depolymerizing bacterium belonged to the genus Vibrio and it was named Vibrio sp. O2. The alginate lyase genes of Vibrio sp. O2 were cloned and expressed in Escherichia coli. Two alginate lyase-producing clones, pVOA-A4 and pVOA-B5, were obtained. The alginate lyase gene alyVOA from pVOA-A4 was composed of an 858-bp open reading frame (ORF) encoding 285 amino acid residues, while alyVOB from pVOA-B5 was composed of an 828-bp ORF encoding 275 amino acid residues. The degree of identity between the deduced amino acid sequences of AlyVOA or AlyVOB and Photobacterium sp. ATCC43367 alginate poly(ManA)lyase AlxM was 92.3% or 32.6%, respectively. Alginate lyase consensus regions corresponding to the sequences YFKAGXYXQ and RXELR were observed in all three of these sequences. AlyVOA and AlyVOB both degraded polymannuronate in plate assays and were therefore confirmed to be poly(β-D-mannuronate)lyases.

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

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6

Similar content being viewed by others

References

  • AJ Baron TY Wong SJ Hick P Gacesa D Willcock MJ McPherson (1994) ArticleTitleAlginate lyase of Klebsiella pneumoniae subsp. Aerogenes: gene cloning, sequence analysis and high-level production in Escherichia coli Gene 143 61–66 Occurrence Handle10.1016/0378-1119(94)90605-X

    Article  Google Scholar 

  • A Boyd AM Chakrabarty (1994) ArticleTitleRole of alginate lyase in cell detachment of Pseudomonas aeruginosa Appl Environ Microbiol 60 2355–2359

    Google Scholar 

  • F Chavagnat C Duez M Guinand P Potin T Barbeyron B Henrissat J Wallach JM Ghuysen (1996) ArticleTitleCloning, sequencing and overexpression in Escherichia coli of the alginate lyase-encoding aly gene of Pseudomonas alginovora: identification of three classes of alginate lyases Biochem J 319 575–583

    Google Scholar 

  • P Gacesa (1988) ArticleTitleAlginates Carbohydr Polym 8 161–182 Occurrence Handle10.1016/0144-8617(88)90001-X

    Article  Google Scholar 

  • P Gacesa (1992) ArticleTitleEnzymic degradation of alginates Int J Biochem 24 545–552 Occurrence Handle10.1016/0020-711X(92)90325-U

    Article  Google Scholar 

  • P Gacesa FS Wusteman (1990) ArticleTitlePlate assay for simultaneous detection of alginate lyase and determination of substrate specificity Appl Environ Microbiol 56 2265–2267

    Google Scholar 

  • F Han QH Gong K Son JB Li WG Yu (2004) ArticleTitleCloning, sequence analysis and expression of gene alyVI encoding alginate lyase from marine bacterium Vibrio sp. strain QY101 DNA Seq 15 344–350

    Google Scholar 

  • A Haug B Larsen O Smidsrot (1966) ArticleTitleA study of the constitution of alginic acid by partial acid hydrolysis Acta Chem Scand 20 183–190 Occurrence Handle10.3891/acta.chem.scand.20-0183

    Article  Google Scholar 

  • UK Laemmli (1970) ArticleTitleCleavage of structural proteins during the assembly of the head of bacteriophage T4 Nature 227 680–685 Occurrence Handle10.1038/227680a0

    Article  Google Scholar 

  • M Malissard C Duez M Guinand MJ Vacheron G Michel N Marty B Joris I Thamm JM Ghuysen (1993) ArticleTitleSequence of a gene encoding a (poly ManA) alginate lyase active on Pseudomonas aeruginosa alginate FEMS Microbiol Lett 110 101–106 Occurrence Handle10.1111/j.1574-6968.1993.tb06302.x

    Article  Google Scholar 

  • Y Matsubara R Kawada K Iwasaki Y Kimura T Oda T Muramatsu (2000) ArticleTitleCloning and sequence analysis of a gene (alyPG) encoding poly(α-l-guluronate)lyase from Corynebacterium sp. strain ALY-1 J Biosci Bioeng 89 199–202 Occurrence Handle10.1016/S1389-1723(00)88738-1

    Article  Google Scholar 

  • WW Nichols MJ Evans MPE Slack HL Walmsley (1989) ArticleTitleThe penetration of antibiotics into aggregates of mucoid and non-mucoid Pseudomonas aeruginosa J Gen Microbiol 135 1291–1303

    Google Scholar 

  • CH Oppenhimer CE Zobell (1952) ArticleTitleThe growth and viability of sixty-three spiecies of marine bacteria as influenced by hydrostatic pressure J Mar Res 11 10–18

    Google Scholar 

  • A Pecina A Paneque (1994) ArticleTitleDetection of alginate lyase by activity staining after sodium dodecil sulfate-polyacrylamide gel electrophoresis and subsequent renaturation Anal Biochem 217 124–127 Occurrence Handle10.1006/abio.1994.1092

    Article  Google Scholar 

  • T Romeo JF Preston (1985) ArticleTitlePurification and structural properties of an extracellular (1-4)-β-d-mannuronan-specific alginate lyase from marine bacterium Biochemistry 25 8385–8391 Occurrence Handle10.1021/bi00374a009

    Article  Google Scholar 

  • W Sabra A-P Zeng W-D Deckwer (2001) ArticleTitleBacterial alginate: physiology, product quality and prosess aspects Appl Microbiol Biotechnol 56 315–325 Occurrence Handle10.1007/s002530100699

    Article  Google Scholar 

  • J Sambrook DW Russell (2001) Molecular Cloning: A Laboratory Manual EditionNumber3 Cold Spring Harbor Laboratory Press Cold Spring Harbor, NY

    Google Scholar 

  • T Sawabe H Takahashi Y Ezura P Gacesa (2001) ArticleTitleCloning, sequence analysis and expression of Pseudoalteromonas elyakovii IAM 14594 gene (alyPEEC) encoding the extracellular alginate lyase Carbohydr Res 335 11–21 Occurrence Handle10.1016/S0008-6215(01)00198-7

    Article  Google Scholar 

  • G Skjåk-Bræk H Grasdalen B Larsen (1986) ArticleTitleMonomer sequence and acetylation pattern in same bacterial alginates Carbohydr Res 154 239–250 Occurrence Handle10.1016/S0008-6215(00)90036-3

    Article  Google Scholar 

  • I Sugimura T Sawabe Y Ezura (2001) ArticleTitleBacterial alginate: physiology, product quality and prosess aspects Appl Microbiol Biotechnol 56 315–325 Occurrence Handle10.1007/s002530100699

    Article  Google Scholar 

  • M Tako E Yoza S Tohma (2000) ArticleTitleCloning and sequence analysis of Vibrio halioticoli genes encoding three types of polyguluronate lyase Mar Biotechnol 2 65–73

    Google Scholar 

  • M Tako S Kiyuna S Uechi F Hongo (2001) ArticleTitleIsolation and characterization of alginic acid from commercially cultured Nemacystus decipiens (Itomozuku) Biosci Biotechnol Biochem 65 654–657 Occurrence Handle10.1271/bbb.65.654

    Article  Google Scholar 

  • TY Wong LA Preston NL Schiller (2000) ArticleTitleAlginate lyase: review of major sources and enzyme characterization, structure-function analysis, biological roles, and applications Annu Rev Microbiol 54 289–340 Occurrence Handle10.1146/annurev.micro.54.1.289

    Article  Google Scholar 

  • M Yamasaki K Ogura W Hashimoto B Mikami K Murata (2005) ArticleTitleA structural basis for deporimerization of alginate by polysaccharide lyasefamiliy-7 J Mol Biol 352 11–21 Occurrence Handle10.1016/j.jmb.2005.06.075

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Marine Products Kimuraya Co., Ltd., 3307 Watari, Sakaiminato, Tottori, 684-0072, Japan

    Hitoshi Kawamoto, Yasunari Miki & Takayuki Kimura

  2. Department of Applied Bioscience and Biotechnology, Faculty of Life and Environmental Science, Shimane University, 1060 Nishikawatsu, Matsue, 690-8504, Japan

    Hitoshi Kawamoto, Akio Horibe, Katsunori Tanaka, Makoto Kawamukai & Hideyuki Matsuda

  3. Center for Integrated Research in Science, Shimane University, 1060 Nishikawatsu, Matsue, 690-8504, Japan

    Tsuyoshi Nakagawa

Authors
  1. Hitoshi Kawamoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Akio Horibe
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yasunari Miki
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Takayuki Kimura
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Katsunori Tanaka
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Tsuyoshi Nakagawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Makoto Kawamukai
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Hideyuki Matsuda
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Hideyuki Matsuda.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kawamoto, H., Horibe, A., Miki, Y. et al. Cloning and Sequencing Analysis of Alginate Lyase Genes from the Marine Bacterium Vibrio sp. O2. Mar Biotechnol 8, 481–490 (2006). https://doi.org/10.1007/s10126-005-6157-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10126-005-6157-z

Keywords

Search

Navigation