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The Journal of Microbiology

, Volume 49, Issue 1, pp 141–145 | Cite as

Screening and characterization of a cellulase gene from the gut microflora of abalone using metagenomic library

  • Duwoon Kim
  • Se-Na Kim
  • Keun Sik Baik
  • Seong Chan Park
  • Chae Hong Lim
  • Jong-Oh Kim
  • Tai-Sun Shin
  • Myung-Joo Oh
  • Chi Nam SeongEmail author
Articles

Abstract

A metagenomic fosmid library was constructed using genomic DNA isolated from abalone intestine. Screening of a library of 3,840 clones revealed a 36 kb insert of a cellulase positive clone (pAMHElO). A shotgun clone library was constructed using the positive clone (pAMHElO) and further screening of 3,840 shotgun clones with an approximately 5 kb insert size using a Congo red overlay revealed only one cellulase positive clone (pAMHL9). The pAMHL9 consisted of a 5,293-bp DNA sequence and three open reading frames (ORFs). Among the three ORFs, cellulase activity was only shown in the recombinant protein (CelAMll) coded by ORF3, which showed 100% identity with outer membrane protein A from Vibrio alginolyticus 12G01, but no significant sequence homology to known cellulases. The expressed protein (CelAMll) has a molecular weight of approximately 37 kDa and the highest CMC hydrolysis activity was observed at pH 7.0 and 37°C. The carboxymethyl cellulase activity was determined by zymogram active staining and different degraded product profiles for CelAMll were obtained when cellotetraose and cellopentaose were used as the substrates, while no substrate hydrolysis was observed on oligosaccharides such as cellobiose and cellotriose.

Keywords

abalone cellulase metagenomic library gut microflora 

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Copyright information

© The Microbiological Society of Korea and Springer-Verlag Berlin Heidelberg 2011

Authors and Affiliations

  • Duwoon Kim
    • 1
  • Se-Na Kim
    • 2
    • 4
  • Keun Sik Baik
    • 2
  • Seong Chan Park
    • 2
  • Chae Hong Lim
    • 2
  • Jong-Oh Kim
    • 3
  • Tai-Sun Shin
    • 3
  • Myung-Joo Oh
    • 3
  • Chi Nam Seong
    • 2
    Email author
  1. 1.Department of Food Science and Technology and Functional Food Research CenterChonnam National UniversityGwangjuRepublic of Korea
  2. 2.Department of BiologySunchon National UniversitySuncheonRepublic of Korea
  3. 3.Division of Food Science and Aqualife MedicineChonnam National UniversityYeosuRepublic of Korea
  4. 4.Korea Basic Science InstituteSuncheon CenterSuncheonRepublic of Korea

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