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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 H2016DIY-Bio – economic, epistemological and ethical implications and ambivalencesLife Sci Soc Pol12710.1186/s40504-016-0039-1 http://dx.doi.org/10.1186/s40504-016-0039-1
2. Callaway E2015Lab staple agar runs lowNature52817117210.1038/528171a26659158 http://dx.doi.org/10.1038/528171a
3. Giavasis I, Harvey LM, McNeil B2000Gellan gumCrit Rev Biotechnol2017721110.1080/0738855000898416911039329 http://dx.doi.org/10.1080/07388550008984169
4. Pollock TJ1993Gellan-related polysaccharides and the genus SphingomonasJ Gen Microbiol1391939194510.1099/00221287-139-8-1939 http://dx.doi.org/10.1099/00221287-139-8-1939
5. Fialho AM, Moreira LM, Granja AT, Popescu AO, Hoffmann K, Sá-Correia I2008Occurrence, production, and applications of gellan: current state and perspectivesAppl Microbiol Biotechnol7988990010.1007/s00253-008-1496-018506441 http://dx.doi.org/10.1007/s00253-008-1496-0
6. Nampoothiri KM, Singhania RR, Sabarinath C, Pandey A2003Fermentative production of gellan using Sphingomonas paucimobilisProcess Biochem381513151910.1016/S0032-9592(02)00321-7 http://dx.doi.org/10.1016/S0032-9592(02)00321-7
7. Park SF2012Microbiology at home: a short non-laboratory manual for enthusiasts and bioartistsAvailable from: https://exploringtheinvisible.files.wordpress.com/2013/11/manual2013.pdf. Retrieved April 3, 2018
8. Emmert EAB and ASM Task Committee on Laboratory Biosafety2012ASM guidelines for biosafety in teaching laboratoriesAvailable 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 M2014Yellow pigments generation deficient Sphingomonas strain and application thereof in gellan gum productionUS patent8,685,698 B2
10. Kang KS, Veeder GT, Colegrove GT1983Deacetylated polysaccharide S-60US patent4,385,123

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/content/journal/jmbe/10.1128/jmbe.v19i2.1530
2018-06-29
2018-09-26

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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