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Bioengineering 2017, 4(2), 52;

Molecular Diagnostic for Prospecting Polyhydroxyalkanoate-Producing Bacteria

Centro de Ciências Tecnológicas da Terra e do Mar, Universidade do Vale do Itajaí, R. Uruguai 458, 88302-202 Itajaí-SC, Brazil
Department of Soil Science, “Luiz de Queiroz” College of Agriculture, University of São Paulo, Piracicaba-SP 13418-260, Brazil
Author to whom correspondence should be addressed.
Academic Editor: Martin Koller
Received: 24 April 2017 / Revised: 12 May 2017 / Accepted: 19 May 2017 / Published: 25 May 2017
(This article belongs to the Special Issue Advances in Polyhydroxyalkanoate (PHA) Production)
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The use of molecular diagnostic techniques for bioprospecting and microbial diversity study purposes has gained more attention thanks to their functionality, low cost and quick results. In this context, ten degenerate primers were designed for the amplification of polyhydroxyalkanoate synthase (phaC) gene, which is involved in the production of polyhydroxyalkanoate (PHA)—a biodegradable, renewable biopolymer. Primers were designed based on multiple alignments of phaC gene sequences from 218 species that have their genomes already analyzed and deposited at Biocyc databank. The combination of oligos phaCF3/phaCR1 allowed the amplification of the expected product (PHA synthases families types I and IV) from reference organisms used as positive control (PHA producer). The method was also tested in a multiplex system with two combinations of initiators, using 16 colonies of marine bacteria (pre-characterized for PHA production) as a DNA template. All amplicon positive organisms (n = 9) were also PHA producers, thus no false positives were observed. Amplified DNA was sequenced (n = 4), allowing for the confirmation of the phaC gene identity as well its diversity among marine bacteria. Primers were also tested for screening purposes using 37 colonies from six different environments. Almost 30% of the organisms presented the target amplicon. Thus, the proposed primers are an efficient tool for screening bacteria with potential for the production of PHA as well to study PHA genetic diversity. View Full-Text
Keywords: bioprospecting; biopolymer; environmental diversity bioprospecting; biopolymer; environmental diversity

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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

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Montenegro, E.M.S.; Delabary, G.S.; Silva, M.A.C.; Andreote, F.D.; Lima, A.O.S. Molecular Diagnostic for Prospecting Polyhydroxyalkanoate-Producing Bacteria. Bioengineering 2017, 4, 52.

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