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Chryseobacterium ginsengiterrae sp. nov., with Beta-Glucosidase Activity Isolated from Soil of a Ginseng Field

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Abstract

The isolated Chryseobacterium ginsengiterrae sp. nov DCY68T was found to be Gram-negative, aerobic, non-motile, non-flagellate and rod-shaped. Their size was approximately 0.40–0.46 × 1.0–1.27 μm. The colonies were yellow-pigmented, convex, circular and 0.5–1.3 mm in diameter when grown on R2A agar for 2 days. DNA, esculin, skim milk, gelatine, starch, Tween 20, and Tween 80 were hydrolyzed, but not cellulose. The cells grew on R2A, TSA, and NA but not on MacConkey agars. Growth occured at 4–33 °C (optimum, 30 °C), at pH 5.0–8.0 (optimum, pH 6.5), and 0–2.5% NaCl. Nitrate was not reduced to nitrite. Oxidase and catalase activity were positive. Strain DCY68T contained β–glucosidase activity in which ginsenoside Rb1 was enzymatically converted to ginsenoside F2. Analysis of the16S rRNA gene sequence revealed that strain C. ginsengiterrae sp. nov DCY68T belonged to the family Flavobacteriaceae and was most closely related to C. limigenitum SUR2T (97.4%). The genomic DNA G+C content was 42.0 mol%. The predominant quinones were MK-6 (74.5%) and MK-7 (25.5%). The major fatty acids were iso-C15:0, summed feature 3 (containing C16:1 ω7c and/or C16:1 ω6c) and iso-C17:0 3-OH. On the basis of these phenotypic, genotypic and chemotaxonomic studies, strain DCY68T represents a novel species of the genus Chryseobacterium, for which name C. ginsengiterrae sp. nov. is proposed. The type strain is DCY68T (=KCTC 32089T = JCM 18517T).

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Acknowledgements

This research was supported by a Grant from Korea Institute of Planning & Evaluation for Technology in Food, Agriculture, Forestry & Fisheries (KIPET No. 317007-3).

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    Authors and Affiliations

    1. Department of Oriental Medicinal Biotechnology, College of Life Sciences, Kyung Hee University, Giheung-Gu, Yongin-Si, Gyeonggi-do, Republic of Korea

      Jong-Hun Noh, Van-An Hoang, Yeon-Ju Kim, Jong-Pyo Kang & Deok-Chun Yang

    2. Graduate School of Biotechnology and Ginseng Bank, College of Life Sciences, Kyung Hee University, Yongin, 446-701, Republic of Korea

      Deok-Chun Yang

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    Correspondence to Yeon-Ju Kim or Deok-Chun Yang.

    Additional information

    Jong-Hun Noh and Van-An Hoang have contributed equally to this work.

    The GenBank/EMBL/DDBJ Accession Number for the 16S rRNA gene sequence of strain DCY68T is JX141783. The DPD Taxonumber of the type strain DCY68T is TA00221.

    Electronic Supplementary Material

    Below is the link to the electronic supplementary material.

    284_2017_1335_MOESM1_ESM.tif

    Supplementary material 1 (TIFF 9301 kb). Supplementary Fig. S1. Transmission Electron Micrography of strain Chryseobacterium ginsengiterrae DCY68T. Bar, 200 nm.

    284_2017_1335_MOESM2_ESM.tif

    Supplementary material 2 (TIFF 3158 kb). Supplementary Fig. S2. Two-dimensional TLC of polar lipid analysis. Polar lipid extracts were obtained from (A), Chryseobacterium ginsengiterrae DCY68T; (B) Ch. aquaticum KCTC 12483T and Ch. defluvii B2T (B), stained with 5% ethanolic molybdophosphoric acid for total polar lipids. Abbreviations: PE, Phosphatidylethanolamine; AL1-5, unidentified polar lipids; L1-4, unidentified polarlipids; APL, unidentified aminophospholipids.

    284_2017_1335_MOESM3_ESM.tif

    Supplementary material 3 (TIFF 1458 kb). Supplementary Fig. S3. Neighbor–joining phylogenetic tree, based on 16S rRNA gene sequences, showing the relationships of strain DCY68T with other Chryseobacterium species. Boot strap values >50% were based on 1000 replications as shown at branching points. Bar, 0.005 substitutions per nucleotide position.

    284_2017_1335_MOESM4_ESM.tif

    Supplementary material 4 (TIFF 433 kb). Supplementary Fig. S4 HPLC analysis of ginsenoside bioconversion Rb1 to ginsenosides F2. A: ginsenosides standard [peak 1, Rg1; 2, Re; 3, Rf; 4, Rg2; 5, Rb1; 6, Rc; 7, Rb2; 8, Rd; 9, F2; 10, Rg3; 11, cK, 12; Rh2], B: control, only substrate Rb1, C: after 1 day incubation at 30 °C, D: after 2 days incubation, E: after 3 days incubation.

    284_2017_1335_MOESM5_ESM.pptx

    Supplementary material 5 (PPTX 64 kb). Supplementary Table S1. Similar characteristics of strain DCY68T and related Chryseobacterium type speciesStrains: 1, DCY68T; 2, Ch. limigenitum SUR2T; 3, Ch. aahli T68T; 4, Ch. aquaticum KCTC 12483T; 5, Ch. soldanellicola PSD1-4T. All data are from this study (+, positive; −, negative).

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    Noh, JH., Hoang, VA., Kim, YJ. et al. Chryseobacterium ginsengiterrae sp. nov., with Beta-Glucosidase Activity Isolated from Soil of a Ginseng Field. Curr Microbiol 74, 1417–1424 (2017). https://doi.org/10.1007/s00284-017-1335-6

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