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Psychrotrophic violacein-producing bacteria isolated from Lake Winnipeg,Canada
Authors:Steven B Kuzyk  Alexander O Pritchard  Jocelyn Plouffe  John L Sorensen  Vladimir Yurkov
Affiliation:1. Microbiology Department, University of Manitoba, R3T 2N2 Winnipeg, MB, Canada;2. Chemistry Department, University of Manitoba, R3T 2N2 Winnipeg, MB, Canada;1. Department of Fisheries and Wildlife, 480 Wilson Road, 13 Natural Resources Building, Michigan State University, East Lansing, MI 48864, United States;2. Department of Microbiology and Molecular Genetics, 567 Wilson Road, 2215 Biomedical Physical Sciences Building, Michigan State University, East Lansing, MI 48864, United States;3. Department of Integrative Biology, 288 Farm Ln, Natural Sciences Building, Michigan State University, East Lansing, MI 48864, United States;1. Department of Biotechnology, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad, 45320, Pakistan;2. Biotechnology and OMICs Lab, Natural and Medical Sciences Research Center, University of Nizwa, P.O. Box 33, Birkatal Al Mauz, Nizwa, 616, Oman;3. Qarshi University, Lahore, Pakistan;1. Environment and Climate Change Canada, 867 Lakeshore Rd., Burlington, Ontario L7S 1A1, Canada;2. Centre for Ecology and Hydrology, Wallingford, Oxfordshire OX10 8BB, UK;3. Department of Biology, University of Waterloo, Waterloo, Ontario, N2L 3G1, Canada;4. Wilfrid Laurier University, Faculty of Science, Biology, 75 University Ave West, Waterloo, ON N2L 3C5, Canada;1. Department of Biology, Lakehead University, 955 Oliver Road, Thunder Bay, ON P7B 5E1, Canada;2. Fisheries and Oceans Canada, Freshwater Institute, 501 University Crescent, MB R3T 2N6, Canada;3. Manitoba Agriculture and Resource Development, 200 Saulteaux Crescent, MB R3J 3W3, Canada;4. IISD-Experimental Lakes Area, Suite 325, 111 Lombard Ave, Winnipeg, MB R3B 0T4, Canada;1. Department of Chemistry, University of Manitoba, Winnipeg, Manitoba R3T 2N2, Canada;2. Department of Physiology, University of Manitoba, Winnipeg, Manitoba R3E 3P5, Canada
Abstract:Violet and purple bacterial pigmentations are uncommon when considering strict aerobes. Here we report discovery of two novel isolates, P102 and P117 from the freshwater Lake Winnipeg, each with violet to deep purple colony colouration. The relationship between pigment production and growth was investigated under different conditions, including a variety of carbon sources, pH, and temperatures. The violet compounds were purified using polar organic solvents, and then structurally characterized via mass spectrometry and nuclear magnetic resonance to be violacein and deoxyviolacein. Strain P117 produced higher concentrations of the darker pigment deoxyviolacein, which resulted in a deeper hue of violet than in cultures of P102. Strain P102, synthesized violacein maximally in liquid rich organic medium at pH 8 and 20 °C, and had 99.3% sequence similarity to the 16S rDNA from Janthinobacterium lividum. Strain P117, related to Massilia violaceinigra sharing 99.2% 16S rDNA sequence similarity, also produced violacein at similar optimal conditions, but developed higher concentrations of pigment at 15 °C. Culture-based methods found that violacein producers composed 0.001% or 0.023% CFU of environmental heterotrophic populations using BG-11 and potato-based media, respectively. Culture-independent high-throughput ribosomal 16S V4 sequencing of environmental DNA was used to detect Gram-negative species known for their production of violacein. Of all bacterial and archaeal sequences present during the fall in littoral waters and sediment of Lake Winnipeg, 5.5% and 6.3%, respectively, belonged to species that have similar pigment, demonstrating the scarcity of violacein producers in this aquatic freshwater system.
Keywords:Bacterial violacein  Bacterial deoxyviolacein  Bacterial community  Psychrophile  Psychrotroph  Lake Winnipeg
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