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Identification of a novel cyclomaltodextrinase annotated as a neopullulanase in the genome of Bacillus cereus

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Abstract

Bacillus cereus is a rod-shaped, gram-positive, motile, and β-hemolytic soil bacterium. B. cereus is an opportunistic pathogen, often responsible for human foodborne illness that is caused by ingestion of starchy foods with symptoms of diarrhea and vomiting. Among the numerous amylolytic enzymes in the genome of the pathogen, the one annotated as a putative neopullulanase (NPase) was cloned and its biochemical properties were characterized in this study. The corresponding gene encoded an enzyme of 586 amino acids with a predicted molecular mass of 68.25 kDa. The putative NPase shared 43.7–59.2% of identity with NPases, cyclomaltodextrinases (CDases), and maltogenic amylases from various bacteria, but shared very low similarity with other amylolytic enzymes of B. cereus. The optimal pH and temperature of the enzyme was 6.5 and 37 ℃, respectively. The enzyme activity was decreased by the cations tested in this study and completely inhibited by Co2+ and Cu2+. The purified enzyme showed substrate preference in the order of α-CD > β-CD > starch > maltodextrin > γ-CD and hydrolyzed them mainly to maltose. However, it did not hydrolyze maltose, pullulan, and glycogen. The enzyme was designated herein as a CDase of B. cereus (BcCDase). Furthermore, the enzyme could transfer the sugars released from CDs and maltotriose to acceptor molecules. BcCDase was likely to be involved in the maltodextrin metabolism in B. cereus.

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Acknowledgements

The authors thank Ms. YB Ko for assistance in collecting reference materials.

Funding

This study was supported by the INU grant in the year of 2019.

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Authors and Affiliations

  1. Department of Life Sciences, Graduate School of Incheon National University, Incheon, 22012, Republic of Korea

    Bo-Ram Park & Jung-Wan Kim

  2. School of Life Sciences, B.S. Abdur Rahman Crescent Institute of Science and Technology, Chennai, 600048, India

    Davoodbasha MubarakAli

  3. Research Center for Bio Material and Process Development, Incheon National University, Incheon, 22012, Republic of Korea

    Davoodbasha MubarakAli & Jung-Wan Kim

  4. Division of Bioengineering, Incheon National University, 12-1 Songdo Dong, Yeonsu Ku, Incheon, 22012, Republic of Korea

    Jung-Wan Kim

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  1. Bo-Ram Park
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B-RP performed investigation, DM prepared the manuscript, and J-WK contributed to funding acquisition, supervision of investigation, and revision of the manuscript.

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Correspondence to Jung-Wan Kim.

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Park, BR., MubarakAli, D. & Kim, JW. Identification of a novel cyclomaltodextrinase annotated as a neopullulanase in the genome of Bacillus cereus. Arch Microbiol 205, 86 (2023). https://doi.org/10.1007/s00203-022-03390-y

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