Pseudomonas carnis isolated from blue discolored fresh cheese and insights into the phylogeny.

Most Pseudomonas spp. are responsible for spoilage in refrigerated foods such as alteration in flavor, texture and appearance. Samples of Minas Frescal cheese with blue discoloration were analysed and contained a high Pseudomonas concentration (7.72 ± 0.36 log CFU/g). Out of the 26 Pseudomonas isolates that were analyzed in our study, 19 demonstrated the capability of producing a diffusible dark pigment. Thus, a pigment-producing isolate (C020) was selected by rep-PCR fingerprinting and subsequently subjected to whole-genome sequencing. The draft genome assembled comprises 42 contigs totaling 6,366,75 bp with an average G + C content of 59.97%, and the species prediction performed by TYGS server, based on the draft genome sequence, identified the C020 as Pseudomonas carnis. In order to investigate the phylogenetic relationships of this isolate with strains already identified of this species, we performed an analysis based on whole-genomic sequences. First, an analysis of all P. carnis genomes deposited in GenBank to date shows that 11% (4/37) are misidentified, and belong to the Pseudomonas paracarnis species. A comparative analysis based on phylogenomic analysis has showed that there is no evolutionary relationship between P. carnis strains carrying second copies of trp genes related to blue discoloration (trpABCDF). This finding reinforces the assertion that these genes are contained in a mobile genetic element. However, it is worth noting that all strains carrying these secondary gene copies have exclusively been isolated from food sources. This observation provides valuable insights into the potential origins and dispersion dynamics of this genetic trait within the species.

The two faces of pyocyanin - why and how to steer its production?

The ambiguous nature of pyocyanin was noted quite early after its discovery. This substance is a recognized Pseudomonas aeruginosa virulence factor that causes problems in cystic fibrosis, wound healing, and microbiologically induced corrosion. However, it can also be a potent chemical with potential use in a wide variety of technologies and applications, e.g. green energy production in microbial fuel cells, biocontrol in agriculture, therapy in medicine, or environmental protection. In this mini-review, we shortly describe the properties of pyocyanin, its role in the physiology of Pseudomonas and show the ever-growing interest in it. We also summarize the possible ways of modulating pyocyanin production. We underline different approaches of the researchers that aim either at lowering or increasing pyocyanin production by using different culturing methods, chemical additives, physical factors (e.g. electromagnetic field), or genetic engineering techniques. The review aims to present the ambiguous character of pyocyanin, underline its potential, and signalize the possible further research directions.

Identification of a novel quinoline-based UV-protective pigment from a psychrotrophic Arctic bacterium.

AIMS: Psychrotrophs are extremophilic microorganisms that grow optimally in low temperature having many unique bioactive molecules of biotechnological applications. In this study, we characterized a pigment from an arctic bacterium with protective activity towards UV exposure. METHODS AND RESULTS: The present research reports isolation and characterization of a psychrotrophic bacteria, RSAP2, from the soil sample of NyAlesund (78°56"N, 11°54"E), Svalbard, Norway. The strain showed closest 16S rRNA gene sequence similarity (99.9%) with Kocuria indica NIO-1021. RSAP2 is a Gram-positive, coccoid aerobe which produces a yellow pigment. The optimal parameters for pigment production while grown in LB medium were 3% (w/v) NaCl and 4 days of incubation of the culture at 20°C and pH 9 with shaking (180 rpm). The pigment was extracted in methanol and acetone (2:1) and further purified through column chromatography. It was characterized by mass spectrometry, UV-visible, fluorescence, IR, 1 H NMR, 13 C NMR spectroscopy and CHNS/O analysis. The pigment has a molecular weight of about 258 daltons and the molecular formula was determined as C15 H18 N2 O2 and is a quinoline derivative. We show that the pigment can protect Escherichia coli against UV-mediated mutagenesis. We further demonstrate that the pigment displays a significant antimicrobial effect and in sublethal concentrations it impairs biofilm formation ability of the model organism Staphylococcus aureus. CONCLUSIONS: The pigment of a psychrotrophic Arctic bacterium, most likely a strain of K. indica, was purified and its chemical structure was determined. The quinoline-based pigment has the ability to protect live cells from UV induced damage. SIGNIFICANCE AND IMPACT OF STUDY: Analysis and characterization of this newly isolated quinoline-based pigment is a potential candidate for future application in skin care products.

Bioactive Pigments from Isolated Bacteria and Its Antibacterial, Antioxidant and Sun Protective Application Useful for Cosmetic Products.

Bacterial pigments are the unique and sustainable source of bioactive colour compounds used in cosmetics, food, textiles, printing and pharmaceutical products. Here, we report the pigment-producing isolates and their biological activities that could be benefited for different industries including cosmeceuticals. In this study, a total of 19 pigment-producing bacteria were isolated and purified from collected soil and water samples. The colour production ability of purified bacteria was observed up to 5 transfers. Of the 19 isolates, two isolates lost colour production ability in subsequent transfers. Crude pigments extracted from the remaining 17 isolates showed sunscreen activity in the range of 0.4-8.34. However, only 6 of them showed significant antibacterial and antioxidant activities. In the media optimization experiment, these 6 bacteria showed optimum growth in neutral to alkaline pH, while optimum temperatures for growth were different for different bacteria. One isolate produces the promising pigment, out of all six potential pigments. It is stable up to 5 transfers, having antioxidant and antibacterial activity with Sun protective activity; the strain was identified using 16srRNA gene sequencing and obtained accession number as MK770403 (probable strain is Staphylococcus xylosus) from National Center for Biotechnology Information (NCBI) database. The results of this study suggested that these bioactive pigments can further be developed and used as antibacterial, antioxidant and sun-protective ingredients in cosmeceuticals.

Purification and characterization of indochrome type blue pigment produced by Pseudarthrobacter sp. 34LCH1 isolated from Atacama desert.

The interest in and demand for natural dyes has increased significantly in recent years; however, very few natural blue dyes are commercially available, because blue colored compounds in nature are relatively rare. In this study, a blue pigment-producing bacteria from Lake Chungará (Atacama Desert, Chile) was isolated, and its blue pigment was purified and chemically characterized. The pigment-producing strain was identified as Pseudarthrobacter sp. by 16S rRNA gene sequencing. The pigment was separated from the filtered culture medium by column chromatography/solid-phase extraction using different resins (ionic exchange, C-18, size exclusion). The strain produced up to 2.5 g L-1 of blue pigment, which was very soluble in water, partially soluble in methanol and insoluble in other organic solvents. The pigment was analyzed and characterized by analytical HPLC, UV-Vis, FT-IR, and H-NMR, and purified by semi-preparative HPLC. The pigment was non-toxic to brine shrimp (LD50 > 2.3 g L-1) and was stable at pH 6-10 at temperatures below 60 °C. HPLC analysis shows that the pigment is composed of four major blue fractions. The physicochemical properties and structural analysis demonstrate that this pigment belongs to the indochrome isomers, whose properties have yet to have been characterized. The high solubility in water, good stability in neutral and basic pH, and negligible toxicity of the blue pigment make it a good candidate suitable for several industrial and possibly some food applications.

Upcycling of food waste streams to valuable biopigments pyocyanin and 1-hydroxyphenazine.

Phenazines, including pyocyanin (PYO) and 1-hydroxyphenazine (1-HP) are extracellular secondary metabolites and multifunctional pigments of Pseudomonas aeruginosa responsible for its blue-green color. These versatile molecules are electrochemically active, involved in significant biological activities giving fitness to the host, but also recognized as antimicrobial and anticancer agents. Their wider application is still limited partly due to the cost of carbon substrate for production, which can be solved by the utilization of carbon from food waste within the biorefinery concept. In this study, a variety of food waste streams (banana peel, potato peel, potato washing, stale bread, yoghurt, processed meat, boiled eggs and mixed canteen waste) was used as sole nutrient source in submerged cultures of P. aeruginosa BK25H. Stale bread was identified as the most suitable substrate to support phenazine biopigments production and bacterial growth. This was further increased in 5-liter fermenter when on average 5.2 mg L-1 of PYO and 4.4 mg L-1 of 1-HP were purified after 24 h batch cultivations from the fermentation medium consisting of homogenized stale bread in tap water. Purified biopigments showed moderate antimicrobial activity, and showed different toxicity profiles, with PYO not being toxic against Caenorhabditis elegans, a free-living soil nematode up to 300 µg mL-1 and 1-HP showing lethal effects at 75 µg mL-1. Therefore, stale bread waste stream with minimal pretreatment should be considered as suitable biorefinery feedstock, as it can support the production of valuable biopigments such as phenazines.

Production, purification, and process optimization of intracellular pigment from novel psychrotolerant Paenibacillus sp. BPW19.

A pink pigment-producing bacterial strain was isolated from wastewater and identified as Paenibacillus sp. BPW19. The motile bacterial strain was Gram-positive, acid fermenting, glucose, sucrose utilizing and rod-shaped with an average cell length of 1.55 µm as studied under the Environmental Scanning Electron Microscope. Even though being psychrotolerant, the cell growth condition of BPW19 was optimized as 25 ºC along with pH 8, and 2.25% inoculum concentration considering the operational ease of the production. Sonication assisted solvent extraction produced 5.41% crude pigment which showed zones of exclusion against gram-negative strains Escherichia coli DH5α, Enterobacter sp. EtK3, and Klebsiella sp. SHC1. Gas Chromatography-Mass Spectrometry analysis of the crude pigment exhibited the dominant presence of major compounds as dotriacontane; 3,7 dimethyl 7 octanal; 1-eicosene and erucic acid. While column chromatography (ethanol:chloroform in 1:4 (v/v) ratio) purified pigment was identified as erucic acid using Nuclear Magnetic Resonance with a net yield of 3.06%.

Draft Genome Sequences and Genomic Analysis for Pigment Production in Bacteria Isolated from Blue Discolored Soymilk and Tofu.

Cold-tolerant bacteria are known to contaminate and cause defects in refrigerated foods. Defects in food products can be observed as changes in appearance, texture, and/or flavor that detract from the product's intended look, feel, or taste. Two distinct organisms were cultured from blue pigmented soymilk and tofu that had been left opened and expired in a home refrigerator. The blue coloration was reproduced when isolates were cultured in fresh, sterile soymilk. These strains also produced a variety of colony color morphologies when cultured on different media types. We report two draft genome sequences of the potential causative agents of blue discoloration of soy foods, Pseudomonas carnis strains UCD_MED3 and UCD_MED7 as well as the 16S rRNA gene sequences of co-occurring strains isolated from the defective soy samples but that did not cause blue discoloration when cultured in fresh soymilk; Serratia liquefaciens strains UCD_MED2 and UCD_MED5.

Extraction and partial characterisation of antioxidant pigment produced by Chryseobacterium sp. kr6.

Pigments synthesised by Chryseobacterium sp. kr6 growing on feather waste were extracted and characterised. The pigment extract was characterised by KOH test, UV-vis, CIELAB colour system, HPLC-DAD-MS, FTIR and its antioxidant capacity was evaluated. A positive bathochromic shift was observed when kr6 colonies or pigment extracts were subjected to alkaline solution (20% KOH) and a λmax at 450 nm was detected for acetone extracts, although no typical fine structure of carotenoids was detected in the electomagnetic spectra. The HPLC profile of the extracted pigment showed that the compound has three different peaks with λmax near 450 nm. The FTIR analysis shows some principal functional groups from a flexirubin-like molecule. The pigmented compound also presents antioxidant activity evaluated by the scavenging of the ABTS radical.

Microbial Production of Violacein and Process Optimization for Dyeing Polyamide Fabrics With Acquired Antimicrobial Properties.

In the present study, crude bacterial extract containing violacein is investigated for the preparation of antimicrobial polyamide fabrics. The optimal culture conditions of Janthinobacterium lividum (JL) for maximum biomass and violacein production were found to be 25°C, pH 7.0, while the addition of ampicillin of 0.2 mg mL-1 in the small scale increased violacein production 1.3-fold. In scale-up trials, the addition of 1% (v/v) glycerol in a fed-batch bioreactor, resulted in fivefold extracted crude violacein increase with final concentration of 1.828 g L-1. Polyamide 6.6 fabrics were dyed following three different processes; through simultaneous fermentation and dyeing (SFD), by incubating the fabric in the sonicated bacterial culture after fermentation and by using cell-free extract containing violacein. Maximum color change (ΔE) and color strength (K/S) obtained for SFD fabrics were 74.81 and 22.01, respectively, while no alteration of fastness and staining of dye at acid and alkaline perspiration or at water was indicated. The dyed fabrics presented significant antifungal activity against Candida albicans, C. parapsilosis, and C. krusei, as well as antibacterial properties against Escherichia coli, Staphylococcus aureus, and the S. aureus MRSA. We have shown that J. lividum cultures can be successfully used for violacein production and for simultaneous dying of fabrics resulting in dyed fabrics with antimicrobial properties without utilization of organic solvents.

Antibacterial activity of the Antarctic bacterium Janthinobacterium sp: SMN 33. 6 against multi-resistant Gram-negative bacteria

Background The increment of resistant strains to commonly used antibiotics in clinical practices places in evidence the urgent need to search for new compounds with antibacterial activity. The adaptations that Antarctic microorganisms have developed, due to the extreme environment that they inhabit, promote them as a potential new source of active compounds for the control of microorganisms causing infections associated with health care. The aim of this study was to evaluate the antibacterial activity of an ethanol extract of the Antarctic bacterium Janthinobacterium sp., strain SMN 33.6, against nosocomial multi-resistant Gram-negative bacteria. Results Inhibitory activity against human Gram-negative bacterial pathogens, with concentrations that varied between 0.5 and 16 µg ml- 1, was demonstrated. Conclusions The ethanolic extract of Janthinobacterium sp. SMN 33.6 possesses antibacterial activity against a chromosomal AmpC beta-lactamase-producing strain of Serratia marcescens, an extended-spectrum beta-lactamase-producing Escherichia coli and also against carbapenemase-producing strains of Acinetobacter baumannii and Pseudomonas aeruginosa. This becomes a potential and interesting biotechnological tool for the control of bacteria with multi-resistance to commonly used antibiotics.