Production Of Carotenoid Pigments Research Articles

Optimization, characterization and biosafety of carotenoids produced from whey using Micrococcus luteus

This study aimed to optimize the production of carotenoid pigments from Micrococcus luteus (ATCC 9341) through the statistical screening of media components and the characterization of antimicrobial, antioxidant, cytogenetic and cytotoxic activities. A BOX-Behnken design was used to assess the effects of whey concentration, inoculum size, pH, temperature, and agitation speed on carotenoid yield. The optimum combination increased production to 2.19 g/L, with a productivity of 0.045 g L-1 h−1 and a productivity yield of 0.644 g/g, as confirmed by an observed carotene production of 2.19 g/L. The final response surface model fitting the data had an R2 of 0.9461. High-performance liquid chromatography (HPLC) analysis identified 12 carotenoid pigment compounds produced by M. luteus. The extracts displayed moderate antimicrobial efficacy against Gram-positive bacteria such as Bacillus cereus (ATCC 11778), Staphylococcus aureus (ATCC 6538), and E. faecalis (ATCC 19433), with inhibition zone diameters (IZD) of 29.0, 14.0, and 37.0 mm, respectively, at 1000 μg/mL. However, its effectiveness against Gram-negative bacteria is limited. In comparison, tetracycline exhibited greater antimicrobial potency. The IC50 value of carotenoids was used to indicate the antioxidant activity. IC50 value from the DPPH assay was 152.80 mg/100mL. An IC50 cytotoxicity value greater than 300 μg/mL was found against normal mouse liver cells, with over 68% cell viability even at 300 μg/mL, indicating low toxicity. Histological structure studies revealed normal myocardial muscle tissue, lung tissue, and kidney tissue sections, whereas liver tissue sections revealed ballooning degeneration of hepatocytes and disorganization of hepatic cords. Cytogenetic parameters revealed that the carotene treatment group had a mitotic index (70%) lower than that of the control but higher than that of the positive control, mitomycin, and did not substantially increase numerical (1.2%) or structural aberrations compared with those of the control, suggesting a lack of genotoxic effects under the experimental conditions. In conclusion, optimized culture conditions enhanced carotenoid yields from M. luteus, and the extracts displayed promising bioactivity as moderate antibiotics against certain gram-positive bacteria and as antioxidants. The high IC50 values demonstrate biosafety. Overall, this bioprocess for enhanced carotenoid production coupled with bioactivity profiling and low cytotoxicity support the application of M. luteus carotenoids.

A novel drying film culture method applying a natural phenomenon: Increased carotenoid production by Haematococcus sp

Low productivity and high cost remain major bottlenecks for the large-scale production of Haematococcus sp. This study explored biomass production and carotenoid accumulation in Haematococcus sp. (KCTC 12348BP) using drying film culture. The broth-cultured strain (3.2 × 106 cells/mL, 0.83 ± 0.02 mg/mL for a 21 d culture) was cultured under various conditions (different inoculum volumes and mist feeding intervals) in waterless agar plates at 28 ± 0.5 °C, under fluorescent light (12 h light–dark cycle) for 1 month. The maximum biomass obtained was 17.60 ± 0.72 g/m2, while the maximum astaxanthin concentration was 8.23 ± 1.13 mg/g in the culture using 1 mL inoculum and 3 d feeding interval. Drought stress in drying film culture effectively induced the accumulation of carotenoids from β-carotene, facilitating the production of canthaxanthin via the astaxanthin biosynthesis pathway. This cost-effective culture system can increase the biomass and carotenoid pigment production in Haematococcus sp.

Study of Some Physiological Activities for Multimetal Tolerant Bacteria Under Metallic Stress

Electroplating effluents contain heavy metals besides nitrate, which can be harmful to aquatic and terrestrial life if released into the environment. Bioremediation is an imperative task to remove these contaminants from aquatic surfaces. Bacterial species that are tolerant to multiple metals were isolated from electroplating effluents and studied for their ability to withstand high metal concentrations and perform physiological activities. These activities included the production of carotenoid pigments and the reduction of nitrate during exponential and stationary growth phases after exposure to high concentrations of single and quaternary metals. The influence of heavy metals on these activities and bacterial behavior was evaluated. The results showed that the production of carotenoid pigments increased over time, with the highest quantities produced at 48 hours with Zn, Cu, and quaternary. Pigments produced vary with the different types of metals and their concentrations. The ratio of pigment extract to unladen metal cells was higher compared to laden metals cells. The isolates were also capable of reducing nitrate and producing nitrite, but high metal concentrations hindered the conversion of nitrite to nitrogen gas and accumulated great amounts of nitrite between the end of the exponential phase and the start of the stationary phase. The production of carotenoids allowed the bacteria to tolerate high metal concentrations and continue reducing nitrate. This study suggests that pigmented bacteria and aerobic denitrifiers have the potential for bioremediation and nitrate reduction, which can help clean and recover contaminated environments.

Gp05, a Prophage-Encoded Virulence Factor, Contributes to Persistent Methicillin-Resistant Staphylococcus aureus Endovascular Infection.

Persistent methicillin-resistant Staphylococcus aureus (MRSA) endovascular infections represent a serious public health threat. We recently demonstrated that the presence of a novel prophage ϕSA169 was associated with vancomycin (VAN) treatment failure in experimental MRSA endocarditis. In this study, we assessed the role of a ϕSA169 gene, ϕ80α_gp05 (gp05), in VAN-persistent outcome using gp05 isogenic MRSA strain sets. Of note, Gp05 significantly influences the intersection of MRSA virulence factors, host immune responses, and antibiotic treatment efficacy, including the following: (i) activity of the significant energy-yielding metabolic pathway (e.g., tricarboxylic acid cycle); (ii) carotenoid pigment production; (iii) (p)ppGpp (guanosine tetra- and pentaphosphate) production, which activates the stringent response and subsequent downstream functional factors (e.g., phenol-soluble modulins and polymorphonuclear neutrophil bactericidal activity); and (iv) persistence to VAN treatment in an experimental infective endocarditis model. These data suggest that Gp05 is a significant virulence factor which contributes to the persistent outcomes in MRSA endovascular infection by multiple pathways. IMPORTANCE Persistent endovascular infections are often caused by MRSA strains that are susceptible to anti-MRSA antibiotics in vitro by CLSI breakpoints. Thus, the persistent outcome represents a unique variant of traditional antibiotic resistance mechanisms and a significant therapeutic challenge. Prophage, a critical mobile genetic element carried by most MRSA isolates, provides their bacterial host with metabolic advantages and resistance mechanisms. However, how prophage-encoded virulence factors interact with the host defense system and antibiotics, driving the persistent outcome, is not well known. In the current study, we demonstrated that a novel prophage gene, gp05, significantly impacts tricarboxylic acid cycle activity, stringent response, and pigmentation, as well as vancomycin treatment outcome in an experimental endocarditis model using isogenic gp05 overexpression and chromosomal deletion mutant MRSA strain sets. The findings significantly advance our understanding of the role of Gp05 in persistent MRSA endovascular infection and provide a potential target for development of novel drugs against these life-threatening infections.

The C-Terminal Acid Phosphatase Module of the RNase HI Enzyme RnhC Controls Rifampin Sensitivity and Light-Dependent Colony Pigmentation of Mycobacterium smegmatis.

RNase H enzymes participate in various processes that require processing of RNA-DNA hybrids, including DNA replication, transcription, and ribonucleotide excision from DNA. Mycobacteria encode multiple RNase H enzymes, and prior data indicate that RNase HI activity is essential for mycobacterial viability. However, the additional roles of mycobacterial RNase Hs are unknown, including whether RNase HII (RnhB and RnhD) excises chromosomal ribonucleotides misincorporated during DNA replication and whether individual RNase HI enzymes (RnhA and RnhC) mediate additional phenotypes. We find that loss of RNase HII activity in Mycobacterium smegmatis (through combined deletion of rnhB/rnhD) or individual RNase HI enzymes does not affect growth, hydroxyurea sensitivity, or mutagenesis, whereas overexpression (OE) of either RNase HII severely compromises bacterial viability. We also show that deletion of rnhC, which encodes a protein with an N-terminal RNase HI domain and a C-terminal acid phosphatase domain, confers sensitivity to rifampin and oxidative stress as well as loss of light-induced carotenoid pigmentation. These phenotypes are due to loss of the activity of the C-terminal acid phosphatase domain rather than the RNase HI activity, suggesting that the acid phosphatase activity may confer rifampin resistance through the antioxidant properties of carotenoid pigment production. IMPORTANCE Mycobacteria encode multiple RNase H enzymes, with RNase HI being essential for viability. Here, we examine additional functions of RNase H enzymes in mycobacteria. We find that RNase HII is not involved in mutagenesis but is highly toxic when overexpressed. The RNase HI enzyme RnhC is required for tolerance to rifampin, but this role is surprisingly independent of its RNase H activity and is instead mediated by an autonomous C-terminal acid phosphatase domain. This study provides new insights into the functions of the multiple RNase H enzymes of mycobacteria.

Genetic Underpinnings of Carotenogenesis and Light-Induced Transcriptome Remodeling in the Opportunistic Pathogen Mycobacterium kansasii.

Mycobacterium kansasii (Mk) causes opportunistic pulmonary infections with tuberculosis-like features. The bacterium is well known for its photochromogenicity, i.e., the production of carotenoid pigments in response to light. The genetics defining the photochromogenic phenotype of Mk has not been investigated and defined pigmentation mutants to facilitate studies on the role of carotenes in the bacterium's biology are not available thus far. In this study, we set out to identify genetic determinants involved in Mk photochromogenicity. We screened a library of ~150,000 transposon mutants for colonies with pigmentation abnormalities. The screen rendered a collection of ~200 mutants. Each of these mutants could be assigned to one of four distinct phenotypic groups. The insertion sites in the mutant collection clustered in three chromosomal regions. A combination of phenotypic analysis, sequence bioinformatics, and gene expression studies linked these regions to carotene biosynthesis, carotene degradation, and monounsaturated fatty acid biosynthesis. Furthermore, introduction of the identified carotenoid biosynthetic gene cluster into non-pigmented Mycobacterium smegmatis endowed the bacterium with photochromogenicity. The studies also led to identification of MarR-type and TetR/AcrR-type regulators controlling photochromogenicity and carotenoid breakdown, respectively. Lastly, the work presented also provides a first insight into the Mk transcriptome changes in response to light.

Studies on optimization of pigment production of SB2 isolate from the Saltern region of Marakanam (T.N)

The marine environment plays a vital role in producing valuable natural products for biomedical and pharmaceutical research. The pigment is one of the bioactive molecules derived from marine bacteria. The strain SB2 was isolated from the saltern areas of Marakanam to produce yellow pigment, round, and smooth colonies on selective media. The strain SB2 was opted out for further studies based on its pigment production (1.68 mg l-1). The incubation period, pH, and temperature are critical factors in increasing pigment production (7.81 mg l-1) from the strain SB2 using RSM (Response Surface Method) by Box Behnken Design (BBD). Similarly, the strain produced the highest pigmentation (7.67 mg l-1) in a nutrient medium containing olive oil, molasses, and corn cob powder. Using BBD, various concentrations of methanol, ethyl ether, and ethanol solvents were optimised with the isolate SB2 to yield maximum pigment (4.29 mg l-1). The extracted pigment was effectively treated with bacterial pathogens to inhibit their growth when the isolate grew at pH 7, temperature 30ºC, and salt 2% concentration. Using Box Behnken Design (BBD), optimising nutrient sources, solvents, and environmental factors with the yellow-pigmented SB2 strain recovered from seawater results in the highest carotenoid pigment production, and the pigment has antibacterial properties.

Experimental determination of radiative characteristics of carotenoid-producing microalgae

Currently, the demand for carotenoid pigments of natural origin has increased due to their diverse application in the industry. One of the natural sources of these compounds are microalgae, which are photosynthetic organisms of high biotechnological value. The production of carotenoid pigments from microalgal cultures are in the scope of sustainability but faces the challenge of optimizing the design and operation of new technologies; biomass production is first maximized and subsequently subjected to stress to induce the synthesis and accumulation of this type of pigments. In this context, the design and scaling of photobioreactors (FBRs) for the cultivation of microalgae is crucial for this type of process. The main objective of the present research work is focused on the application of a new experimental methodology to estimate the radiative characteristics of a microalgae quickly and independently in suspension. Strain Chlorella vulgaris UTEX 2714 (shows high growth rates and accumulate carotenoids such as canthaxanthin and astaxanthin) was used as sample. Extinction coefficient (β), albedo (ω) and asymmetry parameter (g) of the phase function (Φ) were estimated. These parameters define the solution of the radiative transfer equation (RTE) and they are fundamental to characterize the effect of microalgae on radiative transport within any FBR. Both β, and ωwere estimated at different concentrations of microalgae in suspension and were independent of the biomass concentration under the experimental conditions. This study suggests that Φ of Chlorella vulgaris UTEX 2714 can be described using the Henyey-Greenstein model. The radiative characterization of the microalgae was achieved, which is essential for the description of the radiative transport within a FBR and the development of better kinetic models for the scaling of this type of bioprocess.

Identification of major carotenoids from green alga Tetraspora sp. CU2551: partial purification and characterization of lutein, canthaxanthin, neochrome, and β-carotene.

The green algae Tetraspora sp. CU2551 was previously identified as a strain with high potential for biohydrogen production; however, its algal biomass characteristics changed from green to reddish orange within 43days of biohydrogen production. The crude pigments were extracted, partially purified, and characterized by chemical determination. The present study focused on elucidating the carotenoid composition of the selected green alga Tetraspora sp. CU2551. The pigment extract was partially purified and fractionated using thin layer chromatography, and yielded two major and two minor carotenoid bands. The fractions were confirmed by high-performance liquid chromatography with a diode array detector (HPLC-DAD) before being identified and confirmed using Liquid Chromatograph-Quadrupole Time of Flight-Mass Spectrometry (LC-QTOF-MS). The spectral data of these fractions revealed four sub-fractions of interest that were lutein, canthaxanthin, neochrome, and β-carotene, which had percentages in the crude extracts of 30.57%, 25.47%, 7.89%, and 0.71%, respectively. Lutein and canthaxanthin were found to be the major carotenoid pigments present. Our findings in this present study are the first reporting of Tetraspora sp. CU2551 as a potential alternate source for carotenoid pigment production.

Structure and biosynthesis of carotenoids produced by a novel Planococcus sp. isolated from South Africa

BackgroundThe genus Planococcus is comprised of halophilic bacteria generally reported for the production of carotenoid pigments and biosurfactants. In previous work, we showed that the culturing of the orange-pigmented Planococcus sp. CP5-4 isolate increased the evaporation rate of industrial wastewater brine effluent, which we attributed to the orange pigment. This demonstrated the potential application of this bacterium for industrial brine effluent management in evaporation ponds for inland desalination plants. Here we identified a C30-carotenoid biosynthetic gene cluster responsible for pigment biosynthesis in Planococcus sp. CP5-4 through isolation of mutants and genome sequencing. We further compare the core genes of the carotenoid biosynthetic gene clusters identified from different Planococcus species’ genomes which grouped into gene cluster families containing BGCs linked to different carotenoid product chemotypes. Lastly, LC–MS analysis of saponified and unsaponified pigment extracts obtained from cultures of Planococcus sp. CP5-4, revealed the structure of the main (predominant) glucosylated C30-carotenoid fatty acid ester produced by Planococcus sp. CP5-4.ResultsGenome sequence comparisons of isolated mutant strains of Planococcus sp. CP5-4 showed deletions of 146 Kb and 3 Kb for the non-pigmented and “yellow” mutants respectively. Eight candidate genes, likely responsible for C30-carotenoid biosynthesis, were identified on the wild-type genome region corresponding to the deleted segment in the non-pigmented mutant. Six of the eight candidate genes formed a biosynthetic gene cluster. A truncation of crtP was responsible for the “yellow” mutant phenotype. Genome annotation revealed that the genes encoded 4,4′-diapolycopene oxygenase (CrtNb), 4,4′- diapolycopen-4-al dehydrogenase (CrtNc), 4,4′-diapophytoene desaturase (CrtN), 4,4′- diaponeurosporene oxygenase (CrtP), glycerol acyltransferase (Agpat), family 2 glucosyl transferase 2 (Gtf2), phytoene/squalene synthase (CrtM), and cytochrome P450 hydroxylase enzymes. Carotenoid analysis showed that a glucosylated C30-carotenoid fatty acid ester, methyl 5-(6-C17:3)-glucosyl-5, 6′-dihydro-apo-4, 4′-lycopenoate was the main carotenoid compound produced by Planococcus sp. CP5-4.ConclusionWe identified and characterized the carotenoid biosynthetic gene cluster and the C30-carotenoid compound produced by Planococcus sp. CP5-4. Mass-spectrometry guided analysis of the saponified and unsaponified pigment extracts showed that methyl 5-glucosyl-5, 6-dihydro-apo-4, 4′-lycopenoate esterified to heptadecatrienoic acid (C17:3). Furthermore, through phylogenetic analysis of the core carotenoid BGCs of Planococcus species we show that various C30-carotenoid product chemotypes, apart from methyl 5-glucosyl-5, 6-dihydro-apo-4, 4′-lycopenoate and 5-glucosyl-4, 4-diaponeurosporen-4′-ol-4-oic acid, may be produced that could offer opportunities for a variety of applications.

Stability of zeaxanthin/lutein in yolk oil obtained from microalgae-supplemented egg under various storage conditions

Zeaxanthin (Zx) and Lutein (Lut) are yellow-orange carotenoids that are accumulated in the macular region. Dunaliella species are emerging bio-resources for the production of carotenoid pigments. This research demonstrated the potential of D. tertiolecta mp3 mutant to enrich eggs with Zx and Lut and assessed their stability in yolk oil (YO). Laying hens aged 57-week were supplemented with 2–32 g/kg of D. tertiolecta mp3 mutant powder in their diet. Feeding the powder at 0.8–1.6% efficiently increased Zx and Lut contents more than twice and intensified the yolk color as compared to the control group. Enriched YO contained 0.471 mg/g of Zx and 0.640 mg/g of Lut, 315 mg/g of phospholipid and 11.2 mg/g of cholesterol. Subsequently, the effects of temperature (4, 25, 37 and 55 °C), light (884 lux and dark), and atmosphere (ambient air and nitrogen) on the degradation of Zx and Lut were evaluated. As a result, Zx and Lut in YO were well preserved below 25 °C and in dark condition regardless of the presence of air. Consequently, the knowledge of Zx and Lut enrichment and degradation kinetics in YO provides valuable information to the nutraceutical and pharmaceutical industries.

MSI-1 combats drug-resistant S. aureus by affecting bacterial viability and inhibiting carotenoid pigment production

Development of novel therapeutic strategies and antibacterial agents against antibiotic-resistant Staphylococcus aureus (S. aureus) is urgent. In this study, antibacterial activities and possible mechanisms of peptide MSI-1 against multiple drug-resistant S. aureus were investigated. Results demonstrated that MSI-1 had potent bacteriostatic activity and bactericidal efficiency against S. aureus, including methicillin-resistant S. aureus (MRSA), vancomycin-intermediate S. aureus (VISA) and vancomycin-resistant S. aureus (VRSA), with minimum inhibitory concentrations (MICs) ranging from 4 to 16 μg/mL and bactericidal times from 2–12 h. MSI-1 exhibited a low incidence of developing resistance and additive effects with vancomycin to overcome MRSA and VRSA. Moreover, MSI-1, even at sub-MIC concentrations, inhibited staphyloxanthin (STX) production of S. aureus. This inhibitory effect was unique and effectively sensitized S. aureus to host immune defense. In terms of its modes of action, MSI-1 disrupted the cell membrane of S. aureus by binding to negatively-charged lipoteichoic acid to exert a direct bactericidal effect. Interestingly, MSI-1 interacted with 4,4′-diapophytoene desaturase (CrtN) of S. aureus via ionic bonds, hydrogen bonds, and Pi-Pi or Pi-alkyl interactions, and alanine substitution of the key amino acids contributed to these interactions weakened this STX production inhibition. Thus, in a MRSA-induced skin infection in mice and MRSA/VRSA-induced systemic infection in Galleria mellonella,MSI-1 alleviated staphylococcal scalded skin syndrome to promote mouse skin wound repair and mitigated staphylococcus infection-induced immune melanization to enhance G. mellonella survival. Collectively, MSI-1 has potent antibacterial activity against drug-resistant S. aureus by affecting bacterial viability and exerting its anti-virulence effects. It can be developed as a new antibacterial agent to resist refractory S. aureus infection.

Ubiquitousness of Haloferax and Carotenoid Producing Genes in Arabian Sea Coastal Biosystems of India

This study presents a comparative analysis of halophiles from the global open sea and coastal biosystems through shotgun metagenomes (n = 209) retrieved from public repositories. The open sea was significantly enriched with Prochlorococcus and Candidatus pelagibacter. Meanwhile, coastal biosystems were dominated by Marinobacter and Alcanivorax. Halophilic archaea Haloarcula and Haloquandratum, predominant in the coastal biosystem, were significantly (p < 0.05) enriched in coastal biosystems compared to the open sea. Analysis of whole genomes (n = 23,540), retrieved from EzBioCloud, detected crtI in 64.66% of genomes, while cruF was observed in 1.69% Bacteria and 40.75% Archaea. We further confirmed the viability and carotenoid pigment production by pure culture isolation (n = 1351) of extreme halophiles from sediments (n = 410 × 3) sampling at the Arabian coastline of India. All red-pigmented isolates were represented exclusively by Haloferax, resistant to saturated NaCl (6 M), and had >60% G + C content. Multidrug resistance to tetracycline, gentamicin, ampicillin, and chloramphenicol were also observed. Our study showed that coastal biosystems could be more suited for bioprospection of halophiles rather than the open sea.

Pigment Production Improvement in Rhodotorula mucilaginosa AJB01 Using Design of Experiments.

The discovery of biopigments has received considerable attention from the industrial sector, mainly for potential applications as novel molecules with biological activity, in cosmetics or if aquaculture food supplements. The main objective of this study was to increase the production of carotenoid pigments in a naturally pigmented yeast by subjecting the yeast to various cellular stresses using design of experiments. The fungal strain Rhodotorula mucilaginosa AJB01 was isolated from a food sample collected in Barranquilla, Colombia, and one of the pigments produced was β-carotene. This strain was subjected to various stress conditions, including osmotic stress using different salts, physical stress by ultraviolet (UV) light, and light stress using different photoperiods. The optimal growth conditions for carotenoid production were determined to be 1 min of UV light, 0.5 mg/L of magnesium sulfate, and an 18:6 h light/dark period, which resulted in a carotenoid yield of 118.3 µg of carotenoid per gram of yeast.

Enhancement of biomass and total carotenoid content of a UV-resistant strain of Dietzia maris in response to different carbon and nitrogen sources.

In the present study, the effect of various fermentation media on the production of carotenoid pigment in a radiation-resistant strain of Dietzia maris was reported. The biomass and pigment production of this strain was evaluated using various sources of carbon and nitrogen as well as different concentrations of whey medium. The antioxidant and cytotoxic activities of the extracted pigment were also determined using ferric reducing antioxidant power (FRAP), 2,2-diphenyl-1-picrylhydrazyl radicals (DPPH), and MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assays. The antibacterial activity of the carotenoid pigment was also evaluated. All carbon sources increased the pigment production of D. maris in the following order: glucose > raffinose = starch = xylose > sorbitol > sucrose. However, only glucose, xylose, and sorbitol significantly increased the microbial biomass as compared to the control. Moreover, all organic nitrogen sources and ammonium sulfate enhanced the pigment production of the studied strain by approximately 6-9 folds. The free radical scavenging capacity and FRAP of the D. maris carotenoid extract were reported as half-maximal effective concentration or EC50 = 3.30 mg/ml and EC50 =28.46 μg/ml, respectively. The maximum amount of biomass and carotenoid pigment produced by D. maris was obtained in the fermentation medium containing 1 g/l glucose and 1 g/l yeast extract (18 mg/l). This strain can be considered as a promising biocatalyst for the commercial production of natural carotenoids due to its antioxidant capacity and noncytotoxic activity.

Australian Strains of Botryococcus braunii Examined for Potential Hydrocarbon and Carotenoid Pigment Production and the Effect of Brackish Water

The green alga Botryococcus braunii produces abundant hydrocarbons, in the form of drop-in biodiesel, which promoted interest in the species as a renewable fuel. However, despite the observation of dense populations in the wild, Botryococcus grows very slowly in culture, severely limiting its potential for development as a bioresource. Undertaking a biodiscovery program, we found new strains of Botryococcus in locations ranging from tropical to temperate Australia and from both fresh and brackish waters. As part of the ecophysiological characterisation of this new biodiversity, lipid and pigment compositions were studied for six new strains from six different locations. The strains were inoculated in either freshwater or brackish (salinity of 4)-based medium and maintained over 150 days. The growth of cultures was studied continuously, while lipid and pigment composition were analysed at final harvest on day 150. No significant differences in growth rate between fresh and brackish media were observed. Some strains were more tolerable of brackish conditions than others with a link between salinity tolerance and original location. The use of lower salinity (4 ppt) had a minimal effect on lipid composition, with only two of the six strains showing a different hydrocarbon profile in comparison to the other strains; pigment composition showed only minor variations for fresh and brackish water cultures, although the concentrations varied significantly with the freshwater cultures containing higher pigment concentrations.

Use of response surface methodology to enhance carotenoid pigment production from Cellulosimicrobium strain AZ

In the present study, the response surface methodology based on a rotatable central composite design was applied to optimize the production of the carotenoid pigment using a strain of Cellulosimicrobium that has not been reported so far for this genus. The microbial biomass and pigment production of strain AZ were investigated in the presence of tricarboxylic acid cycle intermediates (citrate, malate, succinate), and glutamate. Besides, the influence of the pH of the fermentation medium was also evaluated. The design consisted of a total of 32 experiments at five levels for each factor. Optimum carotenoid production (28.86 mg/L) was observed in the fermentation medium (pH 8.04) containing citrate (11.18 mM), glutamate (12.48 mM), malate (14.19 mM), and succinate (13.38 mM). It was 1.65-fold more than that of the OFAT method (17.5 mg/L) and 12-fold more than the unoptimized conditions (2.4 mg/L). The results were fitted with a quadratic model that could predict the responses to new observations significantly (pred-R2 = 0.9686). Optimum microbial biomass (10.61 g/L) was observed in the presence of citrate (10.27 mM), glutamate (14.03 mM), malate (13.1 mM), and succinate (10.39 mM) as well as pH 8.36. In contrast to the results of one-factor-at-a-time, the carotenoid production had not a direct relationship with bacterial biomass. The established model could describe the variability of above 99.85% in the response based on the determination coefficient (R2).

Carotenoids production in wild yeasts isolated from aquatic systems in Cali, Colombia

Introducción: Los carotenoides son fuente importante de actividades biológicas funcionales, tales como antioxidantes o antimicrobianas, además de tener gran impacto a nivel industrial, ya sea en cosmética o suplementación animal en acuacultura. Se han reportado varias moléculas novedosas a partir de aislamientos en Latinoamérica, principalmente en la Patagonia, Argentina. Sin embargo, no hay reportes en Colombia que evalúen la producción de carotenoides en levaduras nativas pigmentadas. Objetivo: Se evaluó la capacidad de producción de carotenoides en levaduras nativas aisladas de lagos, ríos y aguas residuales de la ciudad de Cali, Colombia. Materiales y métodos: Se caracterizaron 30 levaduras provenientes de dos colecciones. De estas se obtuvo su biomasa, rendimiento de carotenoides totales y producción de β-caroteno. Las cepas promisorias fueron identificadas secuenciando la región ITS1-5.8S-ITS2. Resultados: El mayor rendimiento en la extracción de pigmentos se obtuvo para las cepas P11A (84,36 ± 5,24 µg/g) y Rhodotorula paludigena CS13 (56,26 ± 7,08 µg/g), mientras que las concentraciones más altas de β-caroteno fueron 10,2 µg/mL (R. paludigena CS13) y 9,7 µg/mL (R. mucilaginosa/alborubescens P10A). La cinética de crecimiento y producción de pigmentos durante cinco días fue óptima para la cepa P11A, ya que hubo un aumento en el rendimiento de carotenoides totales 10 veces mayor (48 h: 109,62 µg/g, 120 h: 1403,10 µg/g). Conclusiones: En este estudio se encontró que levaduras aisladas de sistemas acuáticos son promisorias para la producción de pigmentos carotenoides (incluyendo β-caroteno), siendo su extracción y caracterización viable para futuros estudios biotecnológicos.

Reutilization of residual glycerin for the produce β-carotene by Rhodotorula minuta.

This study aimed to evaluate the production of carotenoid pigments by Rhodotorula spp. in submerged fermentation, using residual glycerin from biodiesel production as a carbon source. Chromatographic analysis by HPLC showed that the residual glycerin used as substrate was 57.88% composed of glycerol. The best growth conditions were found in the fermentation medium composed of residual glycerin at a concentration of 30g/L and pH 9. From all the Rhodotorula strains tested, R. minuta URM6693 was selected because of their performance and adaptation in all culture media assayed. The maximum volumetric production of carotenoids was found at 48h (equivalent to 17.20mg/L, for the R. minuta). The production of β-carotene since the first 24h of fermentation reach a final concentration of 1.021mg/L. The yeast Rhodotorula minuta proved its capability to efficiently convert the substrate (mainly at the concentration of 50g/L), obtaining products of biotechnological interest.

Detection the role of physiological factors to produce carotenoid pigment in Staphylococcus aureus

Three Staphylococcus aureus isolates were used in the present study for extraction and quantitation of carotenoid pigment production. Different culture conditions were used to determine optimum pigmentation such as type of culture media, different pH values, temperature, and finally daily bacterial sub-culturing for more than 2 weeks. Nutrient agar was found to be the best medium with the highest production (1.09). Optimum pH value was 6.5 and gave (1.8) carotenoid. Also results showed that pigment was produced at 37o C more than at 42oC. Daily repeated sub culturing had a negative impact on pigment production. Colonies gradualy lost pigmentation and became white in color. Also, sub-culturing was shown to affect the volume of the colonies as they became smaller on solid media.