Carotenoid-producing Bacteria Research Articles

Whole genome sequence of Paracoccus sp. NFXS7, a carotenoid-producing bacterium isolated from a marine saltern.

The study presents the whole genome sequence of the carotenoid-producing Paracoccus sp. NFXS7, isolated from a marine saltern in Setúbal, Portugal. The carotenoid-producing strain NFXS7 contains homologs of the crt genes involved in astaxanthin biosynthesis, making it a promising candidate for biotechnological applications.

Exploring microbial lipases: Screening and identification for biocatalyst potential in bioethanol synthesis from glycerol-based biodiesel waste

Lipases are hydrolytic enzymes that break triglycerides into free fatty acids and glycerol. Typically, these enzymes are produced by microorganisms and used as industrial biocatalysts. Microorganisms isolated from oil-based environmental samples were screened for their lipase-producing ability, collecting twenty bacteria and eighteen fungal strains. Through selective media and morphological identification, various microorganisms were identified, including Pseudomonas sp. (12 strains), Escherichia coli (4 strains), Serratia (2 strains), carotenoid-producing bacteria (1 strain), an unknown isolate bacterium, Aspergillus sp. (6 strains), Aspergillus niger (1 strain), Aspergillus flavus (1 strain), Penicillium sp. (3 strains), Saccharomyces sp. (2 strains), Filamentous Black Fungus (2 strains), Omphalina sp. (1 strain), Rhizopus sp. (1 strain), and Trichoderma sp. (1 strain). Based on lipolytic index results, six bacteria exhibited high lipolytic indices, with Pseudomonas sp. having the highest index. After multi-screening, three lipolytic bacteria belonging to the genera Pseudomonas sp., Serratia sp., and Escherichia coli, as well as three lipolytic fungi belonging to the genera Saccharomyces sp., Trichoderma sp., and Aspergillus flavus, respectively, demonstrated the highest lipolytic activity when screened on a lipase assay, reaching 6.3 and 5.01 mU/g at 72 h of incubation. Consequently, these selected microorganisms were chosen for further study on bioethanol synthesis from glycerol. The study has revealed that the isolated bacteria and fungi strains have the potential to be efficient lipase producers.

A Novel Carotenoid-Producing Bacterium, Paenibacillus aurantius sp. nov., Isolated from Korean Marine Environment.

The novel bacterial strain MBLB1776T was isolated from marine mud in Uljin, the Republic of Korea. Cells were Gram-positive, spore-forming, non-motile, and non-flagellated rods. Growth was observed at a temperature range of 10-45 °C, pH range of 6.0-8.0, and NaCl concentrations of 0-4% (w/v). Phylogenetic analysis of the 16S rRNA gene sequence revealed that MBLB1776T belonged to the genus Paenibacillus and was closely related to Paenibacillus cavernae C4-5T (94.83% similarity). Anteiso-C15:0, iso-C16:0, C16:0, and iso-C15:0 were the predominant fatty acids. Menaquinone 7 was identified as the major isoprenoid quinone. The major polar lipids included diphosphatidylglycerol, phosphatidylglycerol, and phosphatidylethanolamine. Its whole genome was 6.3 Mb in size, with a G+C content of 55.8 mol%. Average nucleotide identity and in silico DNA-DNA hybridization values were below the species delineation threshold. Gene function analysis revealed the presence of a complete C30 carotenoid biosynthetic pathway. Intriguingly, MBLB1776T harbored carotenoid pigments, imparting an orange color to whole cells. Based on this comprehensive polyphasic taxonomy, the MBLB1776T strain represents a novel species within the genus Paenibacillus, for which the name Paenibacillus aurantius sp. nov is proposed. The type strain was MBLB1776T (=KCTC 43279T = JCM 34220T). This is the first report of a carotenoid-producing Paenibacillus sp.

Tsuneonella litorea sp. nov., a novel carotenoid-producing bacterium isolated from coastal sediment.

A Gram-stain-negative, aerobic, non-motile and pleomorphic bacterium designated as YG55T was isolated from a coastal sediment sample. Growth was found to occur at 10-37 °C (optimum, 28 °C), at pH 6-9 (optimum, pH 8) and in 0-6 % NaCl (optimum, 1 %). The results of 16S rRNA gene-based analysis showed that strain YG55T was related to the members of the genus Tsuneonella and shared the highest identity of 99.4 % with Tsuneonella dongtanensis GDMCC 1.2307T, followed by Tsuneonella troitsensis JCM 17037T (98.4 %). The phylogenomic results indicated that strain YG55T formed an independent branch distinct from the reference type strains. The 22.7 and 21.8 % digital DNA-DNA hybridization (dDDH) values and 83.0 and 81.8 % average nucleotide identity (ANI) values between strain YG55T and the two relatives were below the species definition thresholds of 70 % (dDDH) and 95-96 % (ANI), indicating that the strain represents a novel genospecies. The results of chemotaxonomic characterization indicated that the major cellular fatty acids of strain YG55T were summed feature 8 (C18 : 1 ω6c and/or C18 : 1 ω7c), C14 : 0 2OH and C16 : 0; the main polar lipids comprised diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine and sphingoglycolipid; the respiratory quinone was ubiquinone-10. The genomic size and DNA G+C contents were 3.03 Mbp and 66.98 %. The strain contained carotenoid biosynthesis genes and could produce carotenoids. Based on its genotypic and phenotypic characteristics, strain YG55T is concluded to represent a novel species of the genus Tsuneonella, for which the name Tsuneonella litorea sp. nov. is proposed. The type strain is YG55T (=GDMCC 1.2590 T=KCTC 82812T).

Halomonas antri sp. nov., a carotenoid-producing bacterium isolated from surface seawater.

A Gram-negative, moderately halophilic bacterium, designated as strain Y3S6T, was isolated from a surface seawater sample collected from Dongangyoeng cave, Udo-myeon, Jeju-si, Jeju-do, Repulic of Korea. Cells of strain Y3S6T were aerobic, rod-shaped, non-sporulated, yellow, catalase- negative, oxidase-negative and motile with one polar flagellum. Growth of strain Y3S6T occurred at 15-40 °C (optimum: 25-30 °C), at pH 6.0-9.0 (optimum: pH 7.0) and in the presence of 0-13% NaCl (optimum: 1-6 %, w/v). The novel strain was able to produce carotenoids. Its chemotaxonomic and morphological characteristics were consistent with those of members of the genus Halomonas. Phylogenetic analysis of the 16S rRNA gene sequence revealed that strain Y3S6T formed a clade with Halomonas pellis L5T (98.97 %) and Halomonas saliphila LCB169T(98.90%). The average nucleotide identity and digital DNA-DNA hybridization values of strain Y3S6T with the most closely related strains for which whole genomes are publicly available were 82.3-85.2% and 62.8-66.1 %, respectively. The major fatty acids in strain Y3S6T were C16 : 0, C19 : 0 cyclo ω8c and summed feature 8 (composed of C18 : 1 ω7c and/or C18 : 1 ω6c), and the predominant quinone was Q-9. Its polar lipid profile consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, two unidentified phosphoglycolipid, one unidentified phosphoaminoglycolipid and one unidentified phospholipid. The genomic DNA G+C content based on the draft genome sequence was 64.2 mol%. The results of physiological and biochemical tests and 16S rRNA sequence analysis clearly revealed that strain Y3S6T represents a novel species in the genus Halomonas, for which the name Halomonas antri sp. nov. has been proposed. The type strain is Y3S6T (=KACC 21536T=NBRC 114315=TBRC 15164T).

Isolation and selection of carotenoid-producing bacteria from marine environment and determination of carotenoid extraction condition

The carotenoid is a natural pigment group, which is gradual of interest because of its essential biological activities and great production potentials such as using low-cost material and easy to apply on a large scale. For further development of natural carotenoid products, in this study, fourteen yellow, red, and orange pigment-producing marine bacterial strains were isolated from 25 seawater samples collected along coasts of Phu Quoc island (Kien Giang province), Vung Tau City (Ba Ria – Vung Tau province), Can Gio district (Ho Chi Minh city), and Thang Binh district (Quang Nam province). These isolates were identified belong to seven genera Micrococcus, Kocuria, Microbacterium, Brevibacterium, Bacillus, Rhodococcus and Exiguobacterium with 96.7% - 100% of 16S rRNA sequences identity. Based on carotenoid producing ability and UV-absorbance profile, three of the isolates including Micrococcus sp. 64A3a (yellow), Exiguobacterium sp. YT09 (orange) and Rhodococcus enclensis strain RSA3 (red) were selected for investigation of carotenoid extraction conditions. Methanol at concentration of 100% with ratio of 1 g bacterial cells/10 mL methanol is suitable conditions for extraction of red, orange and yellow pigments from the isolates.

Characteristic Evaluation of Various Formulations of Anti-Aging Cream from Carotenoid Extract of Bacterial Symbiont Virgibacillus salarius Strain 19.PP.Sc1.6

Premature aging can be triggered by free radicals from UV rays, since exposure to these rays can cause the skin to experience oxidative stress. Oxidative stress induces intracellular DNA damage, protein denaturation, and lipid peroxidation that lead to cell death. However, cell death can be prevented with antioxidants such as carotenoids, which are among the potential natural compounds for its treatment. Sources of carotenoids include microbial symbionts associated with Sinularia sp., one of which is the bacterium Virgibacillus salarius strain 19.PP.Sc1.6, a carotenoid-producing bacteria. This study aims to explore the utilization of carotenoids from the bacterium V. salarius strain 19.PP.Sc1.6 for the preparation of anti-aging creams. Furthermore, the method employed three formulations (vs, ow, and wo) containing different types of cream tested for stability, and antioxidant and sunscreen abilities. The results obtained established that the carotenoid extract from V. salarius strain 19.PP.Sc1.6 was more stable in the cream vs. the oil-in-water type cream with an anionic emulsifier.

Engineering a Carotenoid-Overproducing Strain of Azospirillum brasilense for Heterologous Production of Geraniol and Amorphadiene.

Escherichia coli and Saccharomyces cerevisiae have been used extensively for heterologous production of a variety of secondary metabolites. Neither has an endogenous high-flux isoprenoid pathway, required for the production of terpenoids. Azospirillum brasilense, a nonphotosynthetic GRAS (generally recognized as safe) bacterium, produces carotenoids in the presence of light. The carotenoid production increases multifold upon inactivating a gene encoding an anti-sigma factor (ChrR1). We used this A. brasilense mutant (Car-1) as a host for the heterologous production of two high-value phytochemicals, geraniol and amorphadiene. Cloned genes (crtE1 and crtE2) of A. brasilense encoding native geranylgeranyl pyrophosphate synthases (GGPPS), when overexpressed and purified, did not produce geranyl pyrophosphate (GPP) in vitro Therefore, we cloned codon-optimized copies of the Catharanthus roseus genes encoding GPP synthase (GPPS) and geraniol synthase (GES) to show the endogenous intermediates of the carotenoid biosynthetic pathway in the Car-1 strain were utilized for the heterologous production of geraniol in A. brasilense Similarly, cloning and expression of a codon-optimized copy of the amorphadiene synthase (ads) gene from Artemisia annua also led to the heterologous production of amorphadiene in Car-1. Geraniol or amorphadiene content was estimated using gas chromatography-mass spectrometry (GC-MS) and GC. These results demonstrate that Car-1 is a promising host for metabolic engineering, as the naturally available endogenous pool of the intermediates of the carotenoid biosynthetic pathway of A. brasilense can be effectively utilized for the heterologous production of high-value phytochemicals.IMPORTANCE To date, the major host organisms used for the heterologous production of terpenoids, i.e., E. coli and S. cerevisiae, do not have high-flux isoprenoid pathways and involve tedious metabolic engineering to increase the precursor pool. Since carotenoid-producing bacteria carry endogenous high-flux isoprenoid pathways, we used a carotenoid-producing mutant of A. brasilense as a host to show its suitability for the heterologous production of geraniol and amorphadiene as a proof-of-concept. The advantages of using A. brasilense as a model system include (i) dispensability of carotenoids and (ii) the possibility of overproducing carotenoids through a single mutation to exploit high carbon flux for terpenoid production.

Enhanced Coproduction of Cell-Bound Zeaxanthin and Secreted Exopolysaccharides by Sphingobium sp. via Metabolic Engineering and Optimized Fermentation.

Zeaxanthin is a value-added carotenoid with wide applications. This study aims to manipulate a generally recognized as safe and carotenoid-producing bacterium, Sphingobium sp., for enhanced production of zeaxanthin and exopolysaccharides. First, whole-genome sequencing and analysis of pathway genes were applied to define the carotenoid pathway in Sphingobium sp. Second, a Sphingobium transformation system was established to engineer metabolite flux into zeaxanthin. By a combination of chemical mutagenesis and removal of bottlenecks of carotenoid biosynthesis via overexpression of three rate-limiting enzymes, the genetically modified Sphingobium DIZ strain produced 21.26 mg/g dry cell weight of zeaxanthin, which was about 4-fold higher than the wild type. Upon optimization of culture conditions, the DIZ strain produced 479.5 mg/L of zeaxanthin with the productivity of 4.99 mg/L/h and 21.9 g/L of exopolysaccharides using a fed-batch fermentation strategy. This study represents the first genetic manipulation of Sphingobium sp., a biotechnologically important bacterium, for high-yield production of value-added metabolites.

Isolation and molecular identification of carotenoid-producing bacteria

Abstract. The production of pigments from bacteria is significant due to the low cost, high yield and ease of extract compared with other sources. Carotenoids are one of the most important pigments with antioxidant properties which are the precursor of vitamin A synthesis and have antibody overproduction ability, anti-tumor activity and inhibitory effect on the cardiovascular disease. The present study aimed to isolate and identify carotenoid-producing bacteria by highperformance liquid chromatography (HPLC) analysis of their carotenoid pigments. Twenty soil samples were collected from different regions of Tehran. After serial dilution each sample was cultured on BHI agar medium and incubated at 37°C. The pigment-producing bacteria were selected for further identification and their pigments were extracted bymethanol. The screening was carried out at two levels: i) selection of the strains by visual color inspection, ii) analysis of the pigment extracts by UV-VIS spectroscopy and HPLC. The isolates were identified by phenotypic methods and their 16S rDNA gene was amplified by PCR method and sequenced. Staphylococcus epidermidis, Micrococcus aloeverae, Citricoccus alkalitolerans, Rhodococcus zopfii, Arthrobacter agilis, Dietzia natronolimnaea and Rhodococcus ruber were identified as carotenoid-producing strains. The highest rate of absorption was observed using UV-VIS analysis in Staphylococcus epidermidis and Dietzia natronolimnaea. The comparison of HPLC analysis with the standard β-carotene curve showed that the carotenoids were beta-carotene. Micro-organisms are a potential source in the production of pigments. In this study we introduced two genera of bacteria (Staphylococcus epidermidis and Dietzia natronolimnaea) with carotenoid-producing ability.

Structural and functional analysis of the carotenoid biosynthesis genes of a Pseudomonas strain isolated from the excrement of Autumn Darter.

There are many reports about carotenoid-producing bacteria and carotenoid biosynthesis genes. In databases for Pseudomonas genome sequences, there are genes homologous to carotenoid biosynthesis genes, but the function of these genes in Pseudomonas has not been elucidated. In this study, we cloned the carotenoid biosynthesis genes from a Pseudomonas sp. strain, named Akiakane, which was isolated from the excrement of the Autumn Darter dragonfly. Using an Escherichia coli functional expression system, we confirmed that the idi, crtE, crtB, crtI, and crtY gene products of the Akiakane strain show predictable catalytic activities. A cluster of six genes was also found, which was comparable to other carotenoid-producing bacteria that belong to the α-Proteobacteria or γ-Proteobacteria class.

Rapid and Selective Screening Method for Isolation and Identification of Carotenoid-Producing Bacteria.

Carotenoids are naturally occurring yellow to red pigments with many biological activities including antioxidant, anticancer, anti-inflammatory, membrane stabilizers, and precursors for vitamin A. These biological activities are linked with many health benefits (e.g., anticarcinogenic activity, prevention of chronic diseases, etc.), which grew the interest of several industrial sectors especially in food, feed, nutraceuticals, cosmetics, and pharmaceutical industries. The production of natural carotenoids from microbial sources such as bacteria can help meet the growing global market of carotenoids estimated at $1.5 billion in 2014 and is expected to reach 1.8 billion in 2019. This chapter demonstrates, step-by-step, the development of a rapid and selective screening method for isolation and identification of carotenoid-producing microorganisms and their carotenoid analysis. This method involves three main procedures: UV treatment, sequencing analysis of 16S rRNA genes, and carotenoids analysis using rapid and effective HPLC-diode array-MS methods.

Draft Genome Sequence of Bacillus marisflavi TF-11T (JCM 11544), a Carotenoid-Producing Bacterium Isolated from Seawater from a Tidal Flat in the Yellow Sea.

Bacillus marisflavi TF-11T (JCM 11544) is a Gram-positive, spore-forming, and carotenoid-producing bacterium isolated from seawater from a tidal flat in the Yellow Sea. Here, we report the first draft genome sequence of B. marisflavi TF-11T, which comprises 4.31 Mb in 11 scaffolds with a G+C content of 48.57%.

Complete Genome Sequence of Rhodococcus sp. B7740, a Carotenoid-Producing Bacterium Isolated from the Arctic Sea.

Rhodococcus sp. B7740 was isolated from Arctic seawater and selected for its capacity to synthesize carotenoids. Here, we report the complete genome sequence of Rhodococcus sp. B7740 to provide the genetic basis for a better understanding of its carotenoid-accumulating capabilities, and we describe the major features of the genome.

Draft Genome Sequence of the Halophilic Bacterium Halobacillus sp. Strain BAB-2008

The Halobacillus sp. strain BAB-2008 is a moderately halophilic, rod-shaped, Gram-positive, orange-pigmented, carotenoid-producing bacterium isolated from saline soil near Zazam-Solar Park Road, Gujarat, India. Here we present the 3.7-Mb genome sequence to provide insights into its functional genomics and potential applications for carotenoid and enzyme production.

ChemInform Abstract: Research on Search of the Carotenoid‐Producing Microorganisms in Marine Area and the Improvement of Production Ratio

AbstractReview: 8 refs.

Research on Search of the Carotenoid-producing Microorganisms in Marine Area and the Improvement of Production Ratio

Carotenoids are liposoluble pigments widely distributed in nature. More than 750 carotenoids are isolated from natural sources, but only a few kinds are used industrially. The production of carotenoid by microorganisms is to be expected, but few carotenoids originate from living things on land. And there is little knowledge about carotenoid-producing microorganisms in the oceans. The possibility still exists of discovering new carotenoid-producing microorganisms. Sunlight is very strong in subtropical regions. The surface of the sea and coral reefs in these regions is a severe environment for growth of microorganisms. While such conditions produce reactive oxygen species, the continuing strong irradiation can also lead to damaging and lethal photo-oxidative reactions. Many undiscovered microorganisms may possess protective mechanisms such as anti-oxidative activities for survival in this environment. This study focused on marine microorganisms inhabiting coral reefs in the Okinawa area, especially carotenoid-producing bacteria possessing anti-oxidative activities. Many carotenoid-producing microorganisms were collected from subtropical ocean areas (a total of 334 strains of pigmented microorganisms), and the chemical composition, some culture conditions and genetic characteristics of the carotenoids from these microorganisms were examined. Furthermore, similar research was performed using some creatures from the ocean surrounding Kochi Prefecture.

Phylogenetic Analysis of Carotenoid-Producing Marine Microorganisms Around the Coral Reefs of the Kerama Islands of Okinawa

Seawater sample from the coral reefs of the Kerama Islands of Okinawa were assessed for the presence of carotenoid-producing bacteria. Results of 16S rDNA analysis of the bacteria obtained from the isolated bacteria showed unique patterns that were different from those of the bacteria obtained from the ordinary marine area. Phylogenetic analysis revealed a slight correlation with the statistical analysis of the PDA chart patterns. The results suggest that useful materials for human health such as carotenoids can be extracted from many carotenoid-producing bacteria such as those found the coral reefs the Kerama Islands.

Unique diversity of carotenoid-producing bacteria isolated from Misasa, a radioactive site in Japan

We obtained carotenoid-producing microorganisms at a high frequency from water samples collected at Misasa (Tottori, Japan), a region known for its high natural radioactivity content. A comprehensive 16S rRNA gene-based phylogenetic analysis revealed that the 104 potential carotenoid producers isolated from Misasa could be classified into 38 different species belonging to seven bacterial classes (Flavobacteria, Sphingobacteria, alpha-Proteobacteria, gamma-Proteobacteria, Deinococci, Actinobacteria, and Bacilli). Of these 38 species, 14 showed sequence similarities less than 97% to their closest identified relatives, and 9 were related to genera that have not been described earlier in terms of carotenoid production. The red-pigmented isolates belonging to Deinococci showed marked resistance to gamma rays and UV irradiation, while those related to Sphingomonas showed weak resistance. The carotenoids produced by the isolates were zeaxanthin (6 strains), dihydroxyastaxanthin (24 strains), astaxanthin (27 strains), canthaxanthin (10 strains), and unidentified molecular species that were produced by the isolates related to Deinococcus, Exiguobacterium, and Flectobacillus. UV irradiation would be useful for the selective isolation of carotenoid-producing microorganisms, and that new microbial producers and other molecular species of carotenoids may potentially be identified from radioactive environments.

Sphingomonas jaspsi sp. nov., a novel carotenoid-producing bacterium isolated from Misasa, Tottori, Japan

A yellow-pigmented, Gram-negative, motile, strictly aerobic, pleomorphic bacterium (strain TDMA-16(T)) was isolated from a freshwater sample collected at Misasa (Tottori, Japan). Strain TDMA-16(T) was slightly tolerant to gamma-ray irradiation and produced carotenoids, including zeaxanthin, nostoxanthin and an unknown carotenoid, effectively [1.7 mg (g dry cells)(-1)]. The DNA G+C content of strain TDMA-16(T) was 63.3 mol%. Phylogenetic analysis based on 16S rRNA gene sequences placed strain TDMA-16(T) in a distinct lineage in the family Sphingomonadaceae; sequence data showed that strain TDMA-16(T) was most closely related to Sphingomonas mali IFO 15500(T) (95.1 %), Sphingomonas aquatilis JSS7(T) (95.0 %), Sphingomonas pruni IFO 15498(T) (94.9 %), Sphingomonas melonis DSM 14444(T) (94.9 %) and Sphingomonas asaccharolytica IFO 15499(T) (94.5 %). The major fatty acids of strain TDMA-16(T) were C(17 : 1) omega 6c (34.5 %) and C(18 : 1) omega 7c (29.3 %). The presence of Q-10 as the main ubiquinone, the Sphingomonadaceae-specific sphingoglycolipid in the polar lipid profile and 2-hydroxy fatty acids, plus the absence of 3-hydroxy fatty acids, supported identification of this strain as a member of the genus Sphingomonas sensu stricto. Phylogenetic distinctiveness and unique phenotypic characteristics differentiated strain TDMA-16(T) from closely related Sphingomonas species. The results of polyphasic taxonomic analyses suggest that strain TDMA-16(T) represents a novel Sphingomonas species, for which the name Sphingomonas jaspsi sp. nov. is proposed. The type strain is strain TDMA-16(T) (=NBRC 102120(T)=DSM 18422(T)=CCUG 53607(T)).