Browse > Article
http://dx.doi.org/10.4014/jmb.1907.07045

A Novel Nucleic Lateral Flow Assay for Screening phaR-Containing Bacillus spp.  

Wint, Nay Yee (Department of Biochemistry, Faculty of Pharmacy, Mahidol University)
Han, Khine Kyi (Department of Pharmacology, Faculty of Pharmacy, Mahidol University)
Yamprayoonswat, Wariya (Chulabhorn Graduate Institute, Chulabhorn Royal Academy)
Ruangsuj, Pattarawan (Chulabhorn Graduate Institute, Chulabhorn Royal Academy)
Mangmool, Supachoke (Department of Pharmacology, Faculty of Pharmacy, Mahidol University)
Promptmas, Chamras (Department of Biomedical Engineering, Faculty of Engineering, Mahidol University)
Yasawong, Montri (Department of Biochemistry, Faculty of Pharmacy, Mahidol University)
Publication Information
Journal of Microbiology and Biotechnology / v.31, no.1, 2021 , pp. 123-129 More about this Journal
Abstract
Polyhydroxyalkanoate (PHA) synthase is a key enzyme for PHA production in microorganisms. The class IV PHA synthase is composed of two subunits: PhaC and PhaR. The PhaR subunit, which encodes the phaR gene, is only present in class IV PHA synthases. Therefore, the phaR gene is used as a biomarker for bacteria that contain a class IV PHA synthase, such as some Bacillus spp. The phaR gene was developed to screen phaR-containing Bacillus spp. The phaR screening method involved two steps: phaR gene amplification by PCR and phaR amplicon detection using a DNA lateral flow assay. The screening method has a high specificity for phaR-containing Bacillus spp. The lowest amount of genomic DNA of B. thuringiensis ATCC 10792 that the phaR screening method could detect was 10 pg. This novel screening method improves the specificity and sensitivity of phaR gene screening and reduces the time and cost of the screening process, which could enhance the opportunity to discover good candidate PHA producers. Nevertheless, the screening method can certainly be used as a tool to screen phaR-containing Bacillus spp. from environmental samples.
Keywords
DNA lateral flow; PHA synthase; phaR; PHA; Bacillus;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG. 1997. The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res. 25: 4876-4882.   DOI
2 Tamura K, Stecher G, Peterson D, Filipski A, Kumar S. 2013. MEGA6: Molecular evolutionary genetics analysis version 6.0. Mol. Biol. Evol. 30: 2725-2729.   DOI
3 Page RD. 1996. TreeView: an application to display phylogenetic trees on personal computers. Comput. Appl. Biosci. 12: 357-358.
4 Shamala TR, Chandrashekar A, Vijayendra SVN, Kshama L. 2003. Identification of polyhydroxyalkanoate (PHA)-producing Bacillus spp. using the polymerase chain reaction (PCR). J. Appl. Microbiol. 94: 369-374.   DOI
5 North EJ, Halden RU. 2013. Plastics and environmental health: the road ahead. Rev. Environ. Health 28: 1-8.   DOI
6 Thompson RC, Moore CJ, vom Saal FS, Swan SH. 2009. Plastics, the environment and human health: current consensus and future trends. Philos. Trans. R Soc. Lond B Biol. Sci. 364: 2153-2166.   DOI
7 Urtuvia V, Villegas P, Gonzalez M, Seeger M. 2014. Bacterial production of the biodegradable plastics polyhydroxyalkanoates. Int. J. Biol. Macromol. 70: 208-213.   DOI
8 Rehm BH. 2003. Polyester synthases: natural catalysts for plastics. Biochem. J. 376: 15-33.   DOI
9 Penkhrue W, Khanongnuch C, Masaki K, Pathom-Aree W, Punyodom W, Lumyong S. 2015. Isolation and screening of biopolymerdegrading microorganisms from northern Thailand. World J. Microbiol. Biotechnol. 31: 1431-1442.   DOI
10 Poli A, Di Donato P, Abbamondi GR, Nicolaus B. 2011. Synthesis, production, and biotechnological applications of exopolysaccharides and polyhydroxyalkanoates by archaea. Archaea 2011: 692253.
11 Hyakutake M, Tomizawa S, Mizuno K, Abe H, Tsuge T. 2014. Alcoholytic cleavage of polyhydroxyalkanoate chains by class IV synthases induced by endogenous and exogenous ethanol. Appl. Environ. Microbiol. 80: 1421-1429.   DOI
12 Bergey DH, Holt JG. 1994. Bergey's manual of determinative bacteriology. pp.787. 9th Ed. Philadelphia: Lippincott Williams & Wilkins.
13 Singh M, Patel SK, Kalia VC. 2009. Bacillus subtilis as potential producer for polyhydroxyalkanoates. Microb. Cell Fact. 8: 38.   DOI
14 Skerman VBD, McGowan V, Sneath PHA. 1980. Approved lists of bacterial names. Int. J. Syst. Evol. Microbiol. 30: 225-420.   DOI
15 Parte AC. 2018. LPSN - List of Prokaryotic names with Standing in Nomenclature (bacterio.net), 20 years on. Int. J. Syst. Evol. Microbiol. 68: 1825-1829.   DOI
16 Tsuge T, Hyakutake M, Mizuno K. 2015. Class IV polyhydroxyalkanoate (PHA) synthases and PHA-producing Bacillus. Appl. Microbiol. Biotechnol. 99: 6231-6240.   DOI
17 Valappil SP, Peiris D, Langley GJ, Herniman JM, Boccaccini AR, Bucke C, et al. 2007. Polyhydroxyalkanoate (PHA) biosynthesis from structurally unrelated carbon sources by a newly characterized Bacillus spp. J. Biotechnol. 127: 475-487.   DOI
18 Mohandas SP, Balan L, Jayanath G, Anoop BS, Philip R, Cubelio SS, et al. 2018. Biosynthesis and characterization of polyhydroxyalkanoate from marine Bacillus cereus MCCB 281 utilizing glycerol as carbon source. Int. J. Biol. Macromol. 119: 380-392.   DOI
19 Gouda MK, Swellam AE, Omar SH. 2001. Production of PHB by a Bacillus megaterium strain using sugarcane molasses and corn steep liquor as sole carbon and nitrogen sources. Microbiol. Res. 156: 201-207.   DOI
20 Kumar P, Ray S, Patel SK, Lee JK, Kalia VC. 2015. Bioconversion of crude glycerol to polyhydroxyalkanoate by Bacillus thuringiensis under non-limiting nitrogen conditions. Int. J. Biol. Macromol. 78: 9-16.   DOI
21 Narayanan A, Ramana KV. 2012. Polyhydroxybutyrate production in Bacillus mycoides DFC1 using response surface optimization for physico-chemical process parameters. 3Biotech 2: 287-296.
22 Schlegel HG, Lafferty R, Krauss I. 1970. The isolation of mutants not accumulating poly-β-hydroxybutyric acid. Arch. Mikrobiol. 71: 283-294.   DOI
23 Spiekermann P, Rehm BH, Kalscheuer R, Baumeister D, Steinbuchel A. 1999. A sensitive, viable-colony staining method using Nile red for direct screening of bacteria that accumulate polyhydroxyalkanoic acids and other lipid storage compounds. Arch. Mikrobiol. 171: 73-80.
24 Sangkharak K, Prasertsan P. 2012. Screening and identification of polyhydroxyalkanoates producing bacteria and biochemical characterization of their possible application. J. Gen. Appl. Microbiol. 58: 173-182.   DOI
25 Muangsuwan W, Ruangsuj P, Chaichanachaicharn P, Yasawong M. 2015. A novel nucleic lateral flow assay for screening of PHAproducing haloarchaea. J. Microbiol. Methods 116: 8-14.   DOI
26 Muangsuwan W, Promptmas C, Jeamsaksiri W, Bunjongpru W, Srisuwan A, Hruanun C, et al. 2016. Development of an immunoFET biosensor for the detection of biotinylated PCR product. Heliyon 2: e00188.   DOI