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A novel intracellular dextranase derived from Paenibacillus sp. 598K with an ability to degrade cycloisomaltooligosaccharides

  • Daiki Mizushima
  • Takatsugu Miyazaki
  • Yuh Shiwa
  • Keitarou Kimura
  • Shiho Suzuki
  • Nobuyuki Fujita
  • Hirofumi Yoshikawa
  • Atsuo Kimura
  • Shinichi Kitamura
  • Hiroshi Hara
  • Kazumi FunaneEmail author
Biotechnologically relevant enzymes and proteins
  • 92 Downloads

Abstract

Paenibacillus sp. 598K produces cycloisomaltooligosaccharides (CIs) in culture from dextran and starch. CIs are cyclic oligosaccharides consisting of seven or more α-(1 → 6)-linked-d-glucose residues. The extracellular enzyme CI glucanotransferase (PsCITase), which is the member of glycoside hydrolase family 66, catalyzes the final stage of CI production and produces mainly cycloisomaltoheptaose. We have discovered a novel intracellular CI-degrading dextranase (PsDEX598) from Paenibacillus sp. 598K. The 69.7-kDa recombinant PsDEX598 does not digest isomaltotetraose or shorter isomaltooligosaccharides, but digests longer ones of at least up to isomaltoheptaose. It also digests oligoCIs of cycloisomaltoheptaose, cycloisomaltooctaose, and cycloisomaltononaose better than it does with megaloCIs of cycloisomaltodecaose, cycloisomaltoundecaose, and cycloisomaltododecaose, as well as an α-(1 → 6)-d-glucan of dextran 40. PsDEX598 is produced intracellularly when culture medium is supplemented with cycloisomaltoheptaose or dextran, but not with isomaltooligosaccharides (a mixture of isomaltose, isomaltotriose, and panose), starch, or glucose. The whole genomic DNA sequence of the strain 598K implies that it harbors two genes for enzymes belonging to glycoside hydrolase family 66 (PsCITase and PsDEX598), and PsDEX598 is the only dextranase in the strain. PsDEX598 does not have any carbohydrate-binding modules (CBMs) and has a low similarity (< 30%) with other family 66 dextranases, and the catalytic amino acids of this enzyme are predicted to be Asp191, Asp303, and Glu368. The strain Paenibacillus sp. 598K appears to take up CI-7, so these findings indicate that this bacterium can degrade CIs using a dextranase within the cells and so utilize them as a carbon source for growth.

Keywords

Dextranase Glycoside hydrolase family 66 Cycloisomaltooligosaccharide Paenibacillus 

Notes

Acknowledgments

We would like to thank Dr. M. Kitaoka for advice on this work. We also would like to thank Enago (www.enago.jp) for the English language review.

Funding

This study was supported in part by the Science and Technology Research Promotion Program for Agriculture, Forestry, Fisheries, and Food Industry in Agriculture, Forestry and Fisheries Research Council (26062B) and KAKENHI in Japan Society for the Promotion of Science (26450133).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals.

Supplementary material

253_2019_9965_MOESM1_ESM.pdf (1.7 mb)
ESM 1 (PDF 1.73 mb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Daiki Mizushima
    • 1
    • 2
  • Takatsugu Miyazaki
    • 1
    • 3
  • Yuh Shiwa
    • 4
  • Keitarou Kimura
    • 1
  • Shiho Suzuki
    • 5
  • Nobuyuki Fujita
    • 4
  • Hirofumi Yoshikawa
    • 6
  • Atsuo Kimura
    • 7
  • Shinichi Kitamura
    • 5
  • Hiroshi Hara
    • 7
    • 8
  • Kazumi Funane
    • 1
    • 9
    Email author
  1. 1.Food Research InstituteNational Agriculture and Food Research Organization (NARO)TsukubaJapan
  2. 2.Division of Medical Zoology, Department of Infection and Immunity, School of MedicineJichi Medical UniversityShimotsukeJapan
  3. 3.Research Institute of Green Science and TechnologyShizuoka UniversityShizuokaJapan
  4. 4.Department of Molecular Microbiology, Faculty of Life SciencesTokyo University of AgricultureSetagaya-kuJapan
  5. 5.Center for Research and Development of Bioresources, Organization for Research PromotionOsaka Prefecture UniversitySakaiJapan
  6. 6.Department of Bioscience, Faculty of Life SciencesTokyo University of AgricultureSetagaya-kuJapan
  7. 7.Research Faculty of AgricultureHokkaido UniversitySapporoJapan
  8. 8.Faculty of Human Life SciencesFuji Women’s UniversityIshikariJapan
  9. 9.Faculty of Life and Environmental SciencesUniversity of YamanashiKofuJapan

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