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Bacterial Production of Gellan Gum as a Do-It-Yourself Alternative to Agar

    Authors: Jenna C. McGuffey1,2,‡, Dacia Leon1,‡, Erum Z. Dhanji2, Dennis M. Mishler1,2, Jeffrey E. Barrick1,*
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    Affiliations: 1: Department of Molecular Biosciences, Center for Systems and Synthetic Biology, The University of Texas at Austin, Austin, TX 78712; 2: Freshman Research Initiative, College of Natural Science, The University of Texas at Austin, Austin, TX 78712
    AUTHOR AND ARTICLE INFORMATION AUTHOR AND ARTICLE INFORMATION
    • Received 17 November 2017 Accepted 10 April 2018 Published 29 June 2018
    • ©2018 Author(s). Published by the American Society for Microbiology.
    • [open-access] This is an Open Access article distributed under the terms of the Creative Commons Attribution-Noncommercial-NoDerivatives 4.0 International license (https://creativecommons.org/licenses/by-nc-nd/4.0/ and https://creativecommons.org/licenses/by-nc-nd/4.0/legalcode), which grants the public the nonexclusive right to copy, distribute, or display the published work.

    • *Corresponding author. Mailing address: The University of Texas at Austin, Department of Molecular Biosciences, 2500 Speedway A5000, Austin, TX 78712. Phone: 512-471-3247. Fax: 512-471-2149. E-mail: [email protected].
    • These authors contributed equally.
    Source: J. Microbiol. Biol. Educ. June 2018 vol. 19 no. 2 doi:10.1128/jmbe.v19i2.1530
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    Abstract:

    Lack of access to reagents and equipment can make performing microbiology experiments difficult in K-12 classrooms and do-it-yourself (DIY) science settings. We demonstrate how the bacterium Sphingomonas paucimobilis can be used to synthesize gellan gum, an alternative to agar, with materials available to DIY scientists and educators. The method involves microwaving cultures of S. paucimobilis after a two-step growth procedure, supplementing them with additional media components as necessary, and then pouring plates. Gellan gum produced in the S. paucimobilis culture acts as a solidifying agent and provides a resilient surface that supports growth of microbes, including Escherichia coli and Saccharomyces cerevisiae. This procedure offers a unique opportunity to experiment with microbial production of an extracellular polysaccharide and to cheaply and sustainably source a reagent for research.

References & Citations

1. Keulartz J, van den Belt H 2016 DIY-Bio – economic, epistemological and ethical implications and ambivalences Life Sci Soc Pol 12 7 10.1186/s40504-016-0039-1 http://dx.doi.org/10.1186/s40504-016-0039-1
2. Callaway E 2015 Lab staple agar runs low Nature 528 171 172 10.1038/528171a 26659158 http://dx.doi.org/10.1038/528171a
3. Giavasis I, Harvey LM, McNeil B 2000 Gellan gum Crit Rev Biotechnol 20 177 211 10.1080/07388550008984169 11039329 http://dx.doi.org/10.1080/07388550008984169
4. Pollock TJ 1993 Gellan-related polysaccharides and the genus Sphingomonas J Gen Microbiol 139 1939 1945 10.1099/00221287-139-8-1939 http://dx.doi.org/10.1099/00221287-139-8-1939
5. Fialho AM, Moreira LM, Granja AT, Popescu AO, Hoffmann K, Sá-Correia I 2008 Occurrence, production, and applications of gellan: current state and perspectives Appl Microbiol Biotechnol 79 889 900 10.1007/s00253-008-1496-0 18506441 http://dx.doi.org/10.1007/s00253-008-1496-0
6. Nampoothiri KM, Singhania RR, Sabarinath C, Pandey A 2003 Fermentative production of gellan using Sphingomonas paucimobilis Process Biochem 38 1513 1519 10.1016/S0032-9592(02)00321-7 http://dx.doi.org/10.1016/S0032-9592(02)00321-7
7. Park SF 2012 Microbiology at home: a short non-laboratory manual for enthusiasts and bioartists Available from: https://exploringtheinvisible.files.wordpress.com/2013/11/manual2013.pdf. Retrieved April 3, 2018
8. Emmert EAB and ASM Task Committee on Laboratory Biosafety 2012 ASM guidelines for biosafety in teaching laboratories Available from: www.asm.org/images/asm_biosafety_guidelines-FINAL.pdf. Retrieved April 3, 2018
9. Wu X, Wu R, Li O, Zhu L, Chen Y, Qian C, Chen M 2014 Yellow pigments generation deficient Sphingomonas strain and application thereof in gellan gum production US patent 8,685,698 B2
10. Kang KS, Veeder GT, Colegrove GT 1983 Deacetylated polysaccharide S-60 US patent 4,385,123

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/content/journal/jmbe/10.1128/jmbe.v19i2.1530
2018-06-29
2019-10-21

Abstract:

Lack of access to reagents and equipment can make performing microbiology experiments difficult in K-12 classrooms and do-it-yourself (DIY) science settings. We demonstrate how the bacterium Sphingomonas paucimobilis can be used to synthesize gellan gum, an alternative to agar, with materials available to DIY scientists and educators. The method involves microwaving cultures of S. paucimobilis after a two-step growth procedure, supplementing them with additional media components as necessary, and then pouring plates. Gellan gum produced in the S. paucimobilis culture acts as a solidifying agent and provides a resilient surface that supports growth of microbes, including Escherichia coli and Saccharomyces cerevisiae. This procedure offers a unique opportunity to experiment with microbial production of an extracellular polysaccharide and to cheaply and sustainably source a reagent for research.

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Figures

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FIGURE 1

DIY gellan gum plates. (a) Initial phase of culturing in rich media. GKB medium before inoculation with (left) and after 2 days of growth (right). (b) Second stage of culturing for gellan gum production. DIY-GPM medium before inoculation (left), immediately after inoculation (middle), and after 8 days of growth (right). The arrow points to gellan gum adhering to the side of the flask. (c) DIY gellan gum plates supplemented with marmite support the growth of colonies of (middle), (right), and (not shown)

Source: J. Microbiol. Biol. Educ. June 2018 vol. 19 no. 2 doi:10.1128/jmbe.v19i2.1530
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