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Bromoperoxidase Producing Bacillus spp. Isolated from the Hypobranchial Glands of A Muricid Mollusc Are Capable of Tyrian Purple Precursor Biogenesis

Marine Ecology Research Centre, School of Environment, Science and Engineering, Southern Cross University, Lismore, NSW 2480, Australia
Southern Cross Plant Science, Southern Cross University, Lismore, NSW 2480, Australia
ARC Industrial Transformation Training Centre for Functional Grains, Charles Sturt University, Wagga Wagga, NSW 2650, Australia
Author to whom correspondence should be addressed.
These authors contributed equally to the manuscript.
Mar. Drugs 2019, 17(5), 264;
Received: 11 April 2019 / Revised: 26 April 2019 / Accepted: 30 April 2019 / Published: 3 May 2019
(This article belongs to the Special Issue Marine Bacteria as Sources of Bioactive Compounds)
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The secondary metabolite Tyrian purple, also known as shellfish purple and royal purple, is a dye with historical importance for humans. The biosynthetic origin of Tyrian purple in Muricidae molluscs is not currently known. A possible role for symbiotic bacteria in the production of tyrindoxyl sulphate, the precursor to Tyrian purple stored in the Australian species, Dicathais orbita, has been proposed. This study aimed to culture bacterial symbionts from the purple producing hypobranchial gland, and screen the isolates for bromoperoxidase genes using molecular methods. The ability of bromoperoxidase positive isolates to produce the brominated indole precursor to Tyrian purple was then established by extraction of the culture, and analysis by liquid chromatography–mass spectrometry (LC–MS). In total, 32 bacterial isolates were cultured from D. orbita hypobranchial glands, using marine agar, marine agar with hypobranchial gland aqueous extracts, blood agar, thiosulphate citrate bile salts sucrose agar, and cetrimide agar at pH 7.2. These included 26 Vibrio spp., two Bacillus spp., one Phaeobacter sp., one Shewanella sp., one Halobacillus sp. and one Pseudoalteromonas sp. The two Bacillus species were the only isolates found to have coding sequences for bromoperoxidase enzymes. LC–MS analysis of the supernatant and cell pellets from the bromoperoxidase producing Bacillus spp. cultured in tryptone broth, supplemented with KBr, confirmed their ability to produce the brominated precursor to Tyrian purple, tyrindoxyl sulphate. This study supports a potential role for symbiotic Bacillus spp. in the biosynthesis of Tyrian purple. View Full-Text
Keywords: shellfish purple; bacillus; bromoperoxidase; tyrindoxyl sulphate; whelk shellfish purple; bacillus; bromoperoxidase; tyrindoxyl sulphate; whelk

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Ngangbam, A.K.; Mouatt, P.; Smith, J.; Waters, D.L.E.; Benkendorff, K. Bromoperoxidase Producing Bacillus spp. Isolated from the Hypobranchial Glands of A Muricid Mollusc Are Capable of Tyrian Purple Precursor Biogenesis. Mar. Drugs 2019, 17, 264.

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