Optimization Of Fermentation Medium Research Articles

Optimized Production of Concanamycins Using a Rational Metabolic Engineering Strategy

Plecomacrolides, such as concanamycin and bafilomycin, are potent and specific inhibitors of vacuolar-type ATPase. Concanamycins are 18-membered macrolides with promising therapeutic potential against multiple diseases, including viral infection, osteoporosis, and cancer. Due to the complexity of their total synthesis, the production of concanamycins is only achieved through microbial fermentation. However, the low titers of concanamycin A and its analogs in the native producing strains are a significant bottleneck for scale-up, robust structure-activity relationship studies, and drug development. To address this challenge, we designed a library of engineered Streptomyces strains for the overproduction of concanamycins by combining the overexpression of target regulatory genes with the optimization of fermentation media. Integration of two endogenous regulators from the concanamycin biosynthetic gene cluster (cms) and one heterologous regulatory gene from the bafilomycin biosynthetic gene cluster into the attB site significantly increased production of concanamycin A and its low abundant analog concanamycin B in Streptomyces eitanensis. The highest titers reported to date were observed in the engineered S. eitanensis DHS10676, which produced over 900 mg/L of concanamycin A and 300 mg/L of concanamycin B. Heterologous overexpression of the identified target regulatory genes across a panel of Streptomyces spp., harboring a putative concanamycin biosynthetic gene cluster confirmed its identity, and significantly improved concanamycin A production in all tested strains. Strain engineering, optimization of fermentation, and extraction purification protocols enabled swift access to these structurally complex plecomacrolides for semi-synthetic medicinal chemistry-based approaches. Together, this work established a platform for robust overproduction of concanamycin analogs across species.

Poly-γ-glutamic production by solid-state fermentation of Bacillus natto in ammonia nitrogen movement and soil water retention processes

A high polyglutamic acid (γ-PGA) producing strain of Bacillus natto UV-40–50 was screened by ultraviolet mutagenesis treatment and identified as still belonging to the Bacillus specie. The optimal fermentation medium composition for solid state fermentation (SSF) of B. natto strain UV-40–50 strain was determined by one-way analysis of variance, under which the yield of γ-PGA was 55.19 g/kg, and the presence and molecular weight of γ-PGA in the γ-PGA-purified samples were determined by a series of characterizations. The purification ability of the unseparated solid fermentation product (SFP) on ammonia nitrogen and nitrite in the water column, as well as its effect on soil water retention, germination rate and seedling length of lettuce and cabbage were further investigated. The results showed that the addition of 1 g/m3 SFP could effectively remove more than 60 % of ammonia nitrogen and more than 40 % of nitrite in the water body; the addition of 0.01 % SFP could increase the water retention capacity of cabbage soil by 2.13 times, and increase the water retention capacity of lettuce soil by 12 %; at the same time, the SFP could also significantly increase the germination rate and seedling length of both cabbage and lettuce.

Optimization of Fermentation and Biocontrol Efficacy of Bacillus atrophaeus XHG-1-3m2.

Biological control plays an increasingly important role in various aspects of modern agriculture and forestry. Identifying biocontrol strains with commercial potential for effective disease management is currently a focal point in biological control research. In this study, Bacillus atrophaeus XHG-1-3m2, a strain with significant biocontrol potential against Wilsonomyces carpophilus causing shot hole disease in wild apricots, was developed. The study determined the antibacterial activity of the fermentation broth, the optimal fermentation medium composition and conditions, and explored its effectiveness in controlling Wilsonomyces carpophilus. The optimal fermentation medium for strain XHG-1-3m2 comprises 12.5 g/L yeast extract, 12.5 g/L soy peptone, 10.0 g/L sodium chloride, 1 g/L ammonium chloride, 1 g/L potassium dihydrogen phosphate, 1 g/L disodium hydrogen phosphate, and 0.5 g/L magnesium sulfate heptahydrate. With an initial pH of 7.0, a liquid volume of 40%, an inoculum volume of 3%, and shaking incubation at 28 °C for 24 h, the viable cell count reached 14 × 109 CFU/mL. In vitro and in vivo tests on leaves revealed that the fermentation broth and the biocontrol biofertilizer derived from this strain inhibited the leaf lesions caused by Wilsonomyces carpophilus on wild apricots, achieving inhibition rates of 94.62% and 82.46%, respectively.

On-site cellulase production by Trichoderma reesei RutC-30 to enhance the enzymatic saccharification of ball-milled corn stover

Cellulases are essential for the enzymatic saccharification of lignocellulose. They play a crucial role in breaking down the structure of lignocellulose to obtain fermentable sugars. In this study, we conducted on-site cellulase production by Trichoderma reesei RutC-30 through submerged fermentation. The effects of carbon source, nitrogen source, KH2PO4, and mineral elements on cellulase production were evaluated using the hydrolyzed total sugar concentration of ball-milled corn stover as an indicator. The optimal fermentation medium conditions for cellulase production were determined through orthogonal experimental design analysis. Additionally, by optimizing culture conditions, including inoculation, pH, and bottling volume, we achieved a total sugar concentration of 92.25 g/L. After the optimization, the FPA, CMCA, protein, and total sugar concentration increased by 75.49 %, 18.43 %, 89.71 %, and 17.83 %, respectively. Furthermore, corn stover pretreated by different methods was applied to induce cellulase production. Ball-milled and steam-exploded corn stover was identified as suitable incubation carbon sources with total sugar concentration up to 94.31 g/L. Our work exploits the cellulase induced by lignocellulose and then applies it to lignocellulose, enabling the customization and providing a reference for the production of cellulase with corn stover as an inducer.

Optimization of Cellulase Production from Agri-Industrial Residues by Aspergillus terreus NIH2624

The objective of this study was to assess the cellulase production of four fungi: Aspergillus terreus NIH2624, Aspergillus clavatus NRRL1, Aspergillus versicolor CBS583.65 and Aspergillus phoenicis ATCC3157, under submerged cultivation conditions. When these fungi were cultured in shake flasks using Mandels and Weber’s minimal medium with 1% sugarcane bagasse as a carbon source and 1.8 g/L of rice bran extract as a nitrogen source, A. terreus showed maximum cellulase production (filter paper activity (FPase) 3.35 U/mL; carboxymethyl cellulase activity (CMCase) 1.69 U/mL). Consequently, A. terreus was selected for the optimization study for cellulase production. Among the different tested carbon sources, A. terreus showed higher CMCase activity when it was cultivated on delignified sugarcane bagasse (1.64 U/mL) and higher FPase activity on sugarcane straw (7.95 U/mL). Regarding the nitrogen sources, the maximum FPase activity was observed when using rice bran (FPase, 8.90 U/mL) and soybean meal (FPase, 9.63 U/mL). The optimized fermentation medium (minimal medium with delignified sugarcane bagasse and rice bran as carbon and nitrogen sources, respectively) resulted in an enzymatic cocktail mainly composed of xylanases, with a maximum activity of 1701.85 U/mL for beechwood xylan, 77.12 U/mL for endoglucanase and 21.02 U/mL for cellobiohydrolase. Additionally, the enzymatic cocktail showed efficient activities for β-glucosidase, β-xylanase, arabinofuranosidase and lytic polysaccharide monoxygenases (LPMOs). This cellulase enzyme solution has the potential to efficiently hydrolyze lignocellulosic biomass, producing second-generation sugars in biorefineries.

Optimization of the production of a new milk clotting enzyme by Aspergillus flavus thermophile on agro-industrial residues medium and characterization of the enzyme

This study focused on the optimization of the production of a milk clotting protease by Aspergillus flavus, isolated from a thermal station. The fermentation was carried out on wheat bran as a medium, followed by a partial characterization of the enzyme. The fungal isolate was screened for protease production based on clear zone hydrolysis on skim milk agar, and then it was identified as belonging to the genus Aspergillus based on its morphological characteristics. This identification was confirmed by molecular analyzes with 100% similarity with the A. flavus strain. After 7 days of fermentation, the crude extract is recovered by simple filtration. The production and optimization of the milk coagulant protease using wheat bran and agro-industrial waste as a medium was carried out using a statistical method of experimental design based on the Plackett and Burman matrices. The results showed that the optimal fermentation medium consisted of wheat bran (4% w/w), incubated at a temperature of 30 °C, with a pH of 5.0 after 6 days of fermentation. The enzyme was partially purified by ammonium sulfate precipitation, yielding an estimated proteolytic activity of 2,985 U. The partially purified protease exhibits optimal activity at pH 5 with hemoglobin as a substrate and an optimal temperature of 50 °C. The Lineweaver-Burk plot showed a KM value of 4.83 g/L and a Vm of 1,560 U. This enzyme causes very rapid coagulation of fresh cow’s milk with an optimum at 45 °C, a pH 7 and a concentration of 50 mM of calcium.

Efficient production of pullulan by Aureobasidium pullulans using a multi-objective optimization strategy with orthogonal experimental design coupling artificial neural network and genetic algorithm

Efficient pullulan production has long been a central research focus. This study used maltodextrin as the carbon source for pullulan production by Aureobasidium pullulans fermentation. A hybrid optimization approach, integrating orthogonal experimental design (OED), backpropagation artificial neural network (BP-ANN), and elite strategy non-dominated sequential genetic algorithm-II (NSGA-II), was developed. Range analysis based on OED revealed that MgSO4·7H2O significantly affects production but less impacts molecular weight, while pH notably influences molecular weight with a lesser effect on production, underscoring the need for multi-objective optimization. The BP-ANN model showed strong predictive capabilities, with goodness-of-fit values of 0.984 and 0.980 for production and molecular weight, respectively. Using this model as the fitness function for the optimization algorithm enhanced efficiency. Taking cost factors into account, the BP-ANN-NSGA-II algorithm identified the optimal fermentation medium conditions, resulting in a 6.89 % increase in production, a 368.97 % increase in molecular weight, and a 42.49 % reduction in cost. The maximum comprehensive optimization efficiency is 63.73 %, and the multi-objective optimization is better than the single objective optimization. This method significantly guides the improvement of pullulan fermentation optimization efficiency.

Study on Optimization of Liquid Fermentation Medium and Antitumor Activity of the Mycelium on Phyllopora lonicerae.

Phylloporia lonicerae is an annual fungus that specifically parasitizes living Lonicera plants, offering significant potential for developing new resource food and medicine. However, wild resources and mycelium production of this fungus is limited, and its anti-tumor active ingredients and mechanisms remain unclear, hampering the development of this fungus. Thus, we optimized the fermentation medium of P. lonicerae and studied the anti-tumor activity of its mycelium. The results indicated that the optimum fermentation medium consisted of 2% sucrose, 0.2% peptone, 0.1% KH2PO4, 0.05% MgSO4·7H2O, 0.16% Lonicera japonica petals, 0.18% P fungal elicitor, and 0.21% L. japonica stem. The biomass reached 7.82 ± 0.41 g/l after 15 days of cultivation in the optimized medium, a 142% increase compared with the potato dextrose broth medium, with a 64% reduction in cultivation time. The intracellular alcohol extract had a higher inhibitory effect on A549 and Eca-109 cells than the intracellular water extract, with half-maximal inhibitory concentration values of 2.42 and 2.92 mg/ml, respectively. Graded extraction of the alcohol extract yielded petroleum ether phase, chloroform phase, ethyl acetate phase, and n-butanol phase. Among them, the petroleum ether phase exhibited a better effect than the positive control, with a half-maximal inhibitory concentration of 113.3 μg/ml. Flow cytometry analysis indicated that petroleum ether components could induce apoptosis of Eca-109 cells, suggesting that this extracted component can be utilized as an anticancer agent in functional foods. This study offers valuable technical support and a theoretical foundation for promoting the comprehensive development and efficient utilization of P. lonicerae.

Stepwise increase of fidaxomicin in an engineered heterologous host Streptomyces albus through multi-level metabolic engineering

The anti-Clostridium difficile infection (CDI) drug fidaxomicin is a natural polyketide metabolite mainly produced by Micromonosporaceae, such as Actinoplanes deccanensis, Dactylosporangium aurantiacum, and Micromonospora echinospora. In the present study, we employed a stepwise strategy by combining heterologous expression, chassis construction, promoter engineering, activator and transporters overexpression, and optimization of fermentation media for high-level production of fidaxomicin. The maximum yield of 384 mg/L fidaxomicin was achieved with engineered Streptomyces albus D7-VHb in 5 L-tank bioreactor, and it was approximately 15-fold higher than the native strain Actinoplanes deccanensis YP-1 with higher strain stability and growth rate. This study developed an enhanced chassis strain, and for the first time, achieved the heterologous synthesis of fidaxomicin through a combinatorial metabolic engineering strategy.

Enhancement of doxorubicin production in Streptomyces peucetius by genetic engineering and process optimization

Doxorubicin is an important class of anthracycline antitumor antibiotics produced by Streptomyces peucetius. The doxorubicin fermentation yield of the wild-type strain was very low, so it could not be produced directly by fermentation at an industrial scale due to the high cost. In the present study, S. peucetius SIPI-7-14 was obtained from SIPI-14 through several rounds of doxorubicin resistance screening. Then, the ketoreductase gene dnrU was knocked out to reduce (13S)-13-dihydrodaunorubicin production, and the resistance gene drrC was overexpressed to further enhance resistance to doxorubicin. The resulting engineered strain S. peucetius △U1/drrC produced 1128 mg/L doxorubicin, a 102.1% increase compared to that of SIPI-14. Then, fermentation medium was optimized using the response surface method. In the optimized fermentation medium, the yield of doxorubicin was increased to 1406 mg/L in shake flask on the 7th day. Furthermore, batch culture was carried out in a 10 L fermenter, and the concentration of doxorubicin reached 1461 mg/L after 7 days of culture, which was the highest yield reported to date, indicating the potential for industrial production of doxorubicin by fermentation.

Chemical and biosynthetic potential of Penicillium shentong XL-F41.

Penicillium strains are renowned for producing diverse secondary metabolites with unique structures and promising bioactivities. Our chemical investigations, accompanied by fermentation media optimization, of a newly isolated fungus, Penicillium shentong XL-F41, led to the isolation of twelve compounds. Among these are two novel indole terpene alkaloids, shentonins A and B (1 and 2), and a new fatty acid 3. Shentonin A (1) is distinguished by an unusual methyl modification at the oxygen atom of the typical succinimide ring, a feature not seen in the structurally similar brocaeloid D. Additionally, shentonin A (1) exhibits a cis relationship between H-3 and H-4, as opposed to the trans configuration in brocaeloid D, suggesting a divergent enzymatic ring-expansion process in their respective fungi. Both shentonins A (1) and B (2) also feature a reduction of a carbonyl to a hydroxy group within the succinimide ring. All isolated compounds were subjected to antimicrobial evaluations, and compound 12 was found to have moderate inhibitory activity against Candia albicans. Moreover, genome sequencing of Penicillium shentong XL-F41 uncovered abundant silent biosynthetic gene clusters, indicating the need for future efforts to activate these clusters and unlock the full chemical potential of the fungus.

Enhanced synthesis of S-adenosyl-L-methionine through combinatorial metabolic engineering and Bayesian optimization in Saccharomyces cerevisiae.

S-Adenosyl-L-methionine (SAM) is a substrate for many enzyme-catalyzed reactions and provides methyl groups in numerous biological methylations, and thus has vast applications in the agriculture and medical field. Saccharomyces cerevisiae has been engineered as a platform with significant potential for producing SAM, but the current production has room for improvement. Thus, a method that consists of a series of metabolic engineering strategies was established in this study. These strategies included enhancing SAM synthesis, increasing ATP supply, down-regulating SAM metabolism, and down-regulating competing pathway. After combinatorial metabolic engineering, Bayesian optimization was conducted on the obtained strain C262P6S to optimize the fermentation medium. A final yield of 2972.8mg·L-1 at 36h with 29.7% of the L-Met conversion rate in the shake flask was achieved, which was 26.3 times higher than that of its parent strain and the highest reported production in the shake flask to date. This paper establishes a feasible foundation for the construction of SAM-producing strains using metabolic engineering strategies and demonstrates the effectiveness of Bayesian optimization in optimizing fermentation medium to enhance the generation of SAM.

Improvement of ribonucleic acid production in Cyberlindnera jadinii and optimization of fermentation medium

To enhance the ribonucleic acid (RNA) productivity for industrial applications, this study employed strain screening and medium optimization to improve the content of RNA in Cyberlindnera jadinii. A rapid screening method, combining atmospheric and room temperature plasma mutagenesis, 48-deep-well plates fermentation, and microplate reader detection, was developed. A mutant strain named WB15 with high RNA content was successfully obtained, exhibiting the RNA content of 156 ± 4.5 mg/g DCW, 1.4 times of the starting strain CCTCC AY 92020. Furthermore, Plackett–Burman design and response surface methodology were employed to identify three significant factors (yeast extract, soybean peptone, and KH2PO4) affecting the RNA content. By utilizing the optimal medium composed of 13.43 g/L yeast extract, 12.12 g/L soybean peptone and 2.78 g/L KH2PO4, the RNA content of WB15 further increased to 184 ± 4.9 mg/g DCW. Additionally, the mutant strain WB15 exhibited a greater cellular width compared to AY 92020, along with increased growth rate and single-cell RNA content by 22% and 48.9%, respectively. Perturbations in ribosome assembly, specifically a reduction in the ratio of ribosomal proteins to ribosomal RNA of the large subunit, might indirectly contribute to the higher RNA content in the WB15 strain. Overall, the combination of rapid screening with fermentation medium optimization proved to be an effective approach for improving the RNA content of C. jadinii, thus facilitating the industrial production of RNA.

Application of sequential design for enhanced L-asparaginase synthesis from Ganoderma australe GPC191.

With an increasing demand for L-asparaginase in pharmaceutical and food sectors for its cytostatic and acrylamide-reducing qualities, there's a need to discover novel, highly productive enzyme sources with improved pharmacokinetic profiles. Keeping this in mind, the present study aimed at maximizing the potential of Ganoderma australe GPC191 to produce L-asparaginase by fermentation medium optimization using statistical validation. Of the 11 physicochemical parameters evaluated under submerged fermentation conditions through one-factor-at-a-time approach and Plackett-Burman design, only four parameters (inoculum load, L-asparagine, soybean meal, and initial pH) influenced L-asparaginase production, significantly (p < 0.001). The optimal levels and interaction effects of these on the overall production were further evaluated by the central composite rotatable design of response surface methodology. Post-optimization, 27.34 U/mL was predicted as the maximum activity at pH 7 with 5n inoculum load and 15g/L each of L-asparagine and soybean meal. Experimental validation yielded an activity of 28.52 U/mL, indicating an overall 18.17-fold increase from the unoptimized stage. To our knowledge, this is the first report signifying the L-asparaginase production aptitude of G. australe with sequential statistical validation using agricultural waste, which can serve as a model to enhance its yields, offering a sustainable and cost-effective solution for industrial application.

Development of platensimycin, platencin, and platensilin overproducers by biosynthetic pathway engineering and fermentation medium optimization.

Implementation of a CRISPR-Cas9 system in Streptomyces platensis CB00739 enabled the construction of a suite of designer recombinant strains for the overproduction of platensimycin, platencin, and platensilin, discovery of new diterpene synthase chemistries, and development of platform strains for future diterpenoid biosynthesis engineering.

Vegan grade medium component screening and concentration optimization for the fermentation of the probiotic strain Lactobacillus paracasei IMC 502® using Design of Experiments.

Lactobacillus paracasei IMC502® is a commercially successful probiotic strain. However, there are no reports that investigate growth medium composition in relation to improved biomass production for this strain. The major outcome of the present study is the design and optimization of a growth medium based on vegan components to be used in the cultivation of Lactobacillus paracasei IMC502®, by using Design of Experiments. Besides comparing different carbon sources, the use of plant-based peptones as nitrogen sources was considered. In particular, the use of guar peptone as the main nitrogen source, in the optimization of fermentation media for the production of probiotics, could replace other plant peptones (e.g. potato, rice, wheat, and soy) which are part of the human diet, thereby avoiding an increase in product and process prices. A model with R2 and adjusted R2 values higher than 95% was obtained. Model accuracy was equal to 94.11%. The vegan-optimized culture medium described in this study increased biomass production by about 65% compared to growth on De Man-Rogosa-Sharpe (MRS) medium. Moreover, this approach showed that most of the salts and trace elements generally present in MRS are not affecting biomass production, thus a simplified medium preparation can be proposed with higher probiotic biomass yield and titer. The possibility to obtain viable lactic acid bacteria at high density from vegetable derived nutrients will be of great interest to specific consumer communities, opening the way to follow this approach with other probiotics of impact for human health.

Fermentation medium optimization of Streptomyces sp. as an antifungal agent against the Ganoderma boninensis pathogen in oil palm

Ganoderma boninensis is the most common fungus which attacks oil palm trees. However, a significant percentage of inhibition to the problem is found through the use of Streptomyces sp. The optimization of the Streptomyces sp. fermentation medium growth factors affects the secondary metabolites production. This study aimed to identify the best formulation of carbon and nitrogen sources and the optimum concentration of Streptomyces sp. fermentation medium for antifungal compound production. The results showed that the best sources of carbon and nitrogen were liquid glucose and monosodium glutamate in the inhibition zones of 16.7 mm and 6.3 mm, while the best concentration levels were 20 g/L and 14.19 g/L, respectively. The results of the first optimization showed an inhibition zone response and area (%) of the optimum high‐performance liquid chromatography (HPLC) chromatogram of 24.39 mm and 62.68 percent, respectively. Taking the suggestion of the first optimization, the second optimization produced 15.2 g/L and 8.3 g/L. The predicted value of the inhibition zone was 21.47 mm, and the area (%) of the HPLC chromatogram was 53.44 percent. The validation results showed an inhibition zone response of 22.01 mm and an HPLC chromatogram area (%) of 54.86 percent. The difference between the predicted and validation values was less than 5 percent; the validation value was thus in line with the value predicted by Design Expert 10.0.7. The chemical formula of the probable active compound is that of the cyclo(phenylalanyl‐prolyl) compound.

Valorisation of chewing gum production waste in bioethanol production: a response surface methodology study

ABSTRACT Interest in fast-moving consumer goods (FMCG) is constantly growing all over the world. Chewing gum, which is in the group of pleasant and entertaining snacks preferred by people of all ages, especially children and young people, is one of the most consumed products in the food sector. Due to its remarkable production and the major components, chewing gum waste has the potential as an alternative sugar source for bioethanol production. To evaluate bioethanol production from chewing gum wastes, the sugar was first extracted, and directly fermented into ethanol under different conditions. Saccharomyces cerevisiae (fresh and dry) and Saccharomyces bayanus were used as the yeast. To determine the optimal fermentation medium, the response surface methodology (RSM) study was employed and the fermentation medium variables were tested such as agar, pepton and glucose (APG medium) concentrations. The APG medium in which the maximum bioethanol yield obtained was determined as 5 g/L agar, 10 g/L peptone and 5 g/L glucose. The highest ethanol yield under optimal APG medium was obtained as 9.1-4 g ethanol/L sugar after 72 h by using Saccharomyces cerevisiae (fresh). The obtained results showed that chewing gum wastes can be utilised a potential source for the bioethanol production.

Biogas from lignocellulosic feedstock: current status and challenges.

The organic wastes and residues generated from agricultural, industrial, and domestic activities have the potential to be converted to bioenergy. One such energy is biogas, which has already been included in rural areas as an alternative cooking energy source and agricultural activities. It is produced via anaerobic digestion of a wide range of organic nutrient sources and is an essential renewable energy source. The factors influencing biogas yield, i.e., the various substrate, their characteristics, pretreatment methods involved, different microbial types, sources, and inoculum properties, are analyzed. Furthermore, the optimization of these parameters, along with fermentation media optimization, such as optimum pH, temperature, and anaerobic digestion strategies, is discussed. Novel approaches of bioaugmentation, co-digestion, phase separation, co-supplementation, nanotechnology, and biorefinery approach have also been explored for biogas production. Finally, the current challenges and prospects of the process are discussed in the review.

Strategies for the Microbial Carotenoids Production Competitiveness Improvement

The research and development of carotenoids production have a long history, and interest in this group of pigments has not decreased to this day. Among all existing carotenoids, six are considered industrially important: astaxanthin, β-carotene, lutein, zeaxanthin, canthaxanthin, and lycopene. These carotenoids have a wide range of application and are used as additives in food and beverage, feed, nutraceuticals, pharmaceuticals, and cosmetics due to their bioactive and colour properties. An undisputed leader in the global pigment market is chemically synthesized carotenoids. To a lesser extent, carotenoids derived from natural sources as plants and microorganisms. Currently, the market of natural carotenoids is mainly represented by microalgae Haematococcus pluvialis, Dunaliella salina, Botryococcus braunii, fungus Blakeslea trispora, yeast Phaffia rhodozyma and bacteria Paracoccus carotinifaciens. These microorganisms afford the production of astaxanthin, βcarotene, canthaxanthin, and lycopene. In turn, lutein is obtained by extracting marigold flowers Tagetes eracta L. and there is no other competitive source yet. Therefore, the potential of microorganisms to synthesize and accumulate lutein and other equally important carotenoids in their cells has been actively studied. Several yeast and bacteria species from Rhodosporidium, Rhodotorula, Sporobolomyces, Sphingomonas, Gordonia, and Sphingobacterium genus have a potential to replenish the diversity of sources of industrially important natural pigments, but available technologies still need improving. This paper reviews strategies for increasing of competitiveness of fungal and bacterial carotenoids production. Strategies such as selection of carotenogenic strain, use of low-cost substrates, simultaneous production of carotenoids and other value-added compounds, and optimization of fermentation medium and conditions are considered.