Optimization Of Medium Composition Research Articles

A Comprehensive Study on the Micropropagation of Costus igneus: Media Composition, Growth, and Development

Costus igneus, commonly known as insulin plant, is an ornamental plant valued for its spiral ginger- like foliage and medicinal properties. Micropropagation is an efficient method for rapid multiplication of Costus igneus. This study aims to determine the optimal media composition and culture conditions for in vitro propagation of C. igneus. Axillary bud explants were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations and combinations of plant growth regulators including auxins (indole-3-acetic acid, indole-3-butyric acid and naphthaleneacetic acid) and cytokinins (6- benzylaminopurine and kinetin). Microshoots cultured on MS + 1.5 mg/L BAP showed the highest shoot proliferation rate (98%) and maximum number of shoots per explant (12.6). Elongated microshoots were rooted on half-strength MS medium supplemented with different auxins. The highest rooting percentage (95%) and maximum number of roots per shoot (9.8) were observed on medium containing 2.5 mg/L IBA. The regenerated plantlets were acclimatized and successfully transferred to pots with 80% survival rate. Morphological and phytochemical analysis showed no significant differences between the in vitro propagated and mother plants. This study demonstrates the potential of micropropagation for large scale production of quality planting material of C. igneus. Further studies on genetic and epigenetic stability are recommended to validate this protocol for commercial applications.

Two-pot optimization of nutrient sources to enhance antioxidants in Citrus reticulata peel powder through solid-state fermentation with Aspergillus niger CGMCC 3.6189

This study aimed to optimize nutrient sources for enhancing the accumulation of antioxidants in Citrus reticulata peel powder (CRPP) through solid-state fermentation (SSF) using Aspergillus niger CGMCC 3.6189. A Plackett-Burman design revealed that four medium compositions, including yeast extract, NH4Cl, CaCl2, and MgCl2, significantly affected antioxidant accumulation in fermented CRPP. The optimal concentrations of these four compositions were predicted as 13.86, 0.40, 0.06, and 0.04 mg/g of CRPP, respectively, using a Box-Behnken design (BBD) when simultaneous optimization of total phenolic content (TPC), total flavonoid content (TFC), 2,2′-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging capacities using Derringer's desirability function. Under the optimal medium compositions, TPC, TFC, ABTS, and DPPH scavenging capacities of fermented CRPP (FO-CRPP) increased by 25.2%, 26.9%, 45.8%, and 57.3%, respectively, in comparison with the unfermented CRPP (UF-CRPP). The HPLC analysis exhibited notable decreases in the levels of hesperidin, narirutin, nobiletin, and tangeretin, while enhancements in the contents of p-coumaric acid, ferulic acid, narirutin, and hesperetin in FO-CRPP compared to UF-CRPP were observed. Scanning electron microscopy and Fourier transform infrared spectroscopy analyses demonstrated changes in the surface microstructure and functional groups in FO-CRPP. Our findings suggested that yeast extract and NH4Cl significantly improve the accumulation of antioxidants in CRPP during SSF by A. niger CGMCC 3.6189.

A primary culture method for the easy, efficient, and effective acquisition of oligodendrocyte lineage cells from neonatal rodent brains

Oligodendrocytes (OL) are myelin-forming glial cells in the central nervous system. In vitro primary OL culture models offer the benefit of a more readily controlled environment that facilitates the examination of diverse OL stages and their intricate dynamics. Although conventional methods for primary OL culture exist, their performance in terms of simplicity and efficiency can be improved. Here, we introduce a novel method for primary OL culture, namely the E3 (easy, efficient, and effective) method, which greatly improves the simplicity and efficiency of the primary OL culture procedure using neonatal rodent brains. We also provided the optimal media composition for the augmentation of oligodendrocyte progenitor cell (OPC) proliferation and more robust maturation into myelin-forming OLs. Overall, E3 offers an undemanding method for obtaining primary OLs with high yield and quality. Alongside its value as a practical tool, in vitro characteristics of the OL lineage additionally identified during the development of the E3 method have implications for advancing research on OL physiology and pathophysiology.

A Thermotolerant Yeast Cyberlindnera rhodanensis DK Isolated from Laphet-so Capable of Extracellular Thermostable β-Glucosidase Production.

This study aims to utilize the microbial resources found within Laphet-so, a traditional fermented tea in Myanmar. A total of 18 isolates of thermotolerant yeasts were obtained from eight samples of Laphet-so collected from southern Shan state, Myanmar. All isolates demonstrated the tannin tolerance, and six isolates were resistant to 5% (w/v) tannin concentration. All 18 isolates were capable of carboxy-methyl cellulose (CMC) degrading, but only the isolate DK showed ethanol production at 45 °C noticed by gas formation. This ethanol producing yeast was identified to be Cyberlindnera rhodanensis based on the sequence analysis of the D1/D2 domain on rRNA gene. C. rhodanensis DK produced 1.70 ± 0.01 U of thermostable extracellular β-glucosidase when cultured at 37 °C for 24 h using 0.5% (w/v) CMC as a carbon source. The best two carbon sources for extracellular β-glucosidase production were found to be either xylose or xylan, with β-glucosidase activity of 3.07-3.08 U/mL when the yeast was cultivated in the yeast malt extract (YM) broth containing either 1% (w/v) xylose or xylan as a sole carbon source at 37 °C for 48 h. The optimal medium compositions for enzyme production predicted by Plackett-Burman design and central composite design (CCD) was composed of yeast extract 5.83 g/L, peptone 10.81 g/L and xylose 20.20 g/L, resulting in a production of 7.96 U/mL, while the medium composed (g/L) of yeast extract 5.79, peptone 13.68 and xylan 20.16 gave 9.45 ± 0.03 U/mL for 48 h cultivation at 37 °C. Crude β-glucosidase exhibited a remarkable stability of 100%, 88% and 75% stable for 3 h at 35, 45 and 55 °C, respectively.

Optimization of medium compositions and X-ray irradiation to enhance monacolin K production by Monascus purpureus in submerged fermentation

The secondary metabolite Monacolin K (MK), derived from Monascus, exhibits potential lipid-lowering and anti-tumor effects. In this study, a mutant strain M. purpureus M400–6 was generated through X-ray treatment, resulting in an average MK production of 181.97 mg/L (a 54.20% increase compared to the parent strain M. purpureus M1) over five successive generations. Furthermore, optimization of medium compositions using response surface methodology led to a further enhancement in MK production by the mutant strain to 266.63 mg/L. Moreover, based on the outcomes of single-factor experiments, the addition of L-citrulline demonstrated a significantly higher level of enhancement in MK production. Therefore, a comparative transcriptome analysis was conducted to unveil the impact of L-citrulline on MK biosynthesis in M. purpureus. The data revealed a significant upregulation of glycolysis pathways, while oxidative phosphorylation including the TCA cycle and the electron transport chain, exhibited significant downregulation. Furthermore, the metabolisms of serine, glycine, threonine, alanine, aspartic acid and glutamate were found to be significantly disturbed. Additionally, the expressions of MPs and MK biosynthetic gene cluster showed a significant downregulation and upregulation respectively. Overall, these findings provide insights into enhancing MK production in submerged fermentation and elucidate the impact of L-citrulline on secondary metabolism.

Direct Organogenesis of Citrus Cultivars from Shoot Tip Nodal Segments.

Citrus cultivar improvement via conventional breeding strategies is impeded by factors related to its reproductive biology. The orange is a hybrid between pomelo (Citrus maxima) and mandarin (Citrus reticulata). Among various orange cultivars, Valencia oranges have a bit of bitter tang mixed in with their sweetness, as Navel oranges are, the most widely cultivated citrus species, quite sweeter, and also don't contain any seeds. Tangelo mandarin orange cultivar is a hybrid of C. reticulata × C. maxima or × C. paradisi. The present study was undertaken to optimize the hormonal composition of the media with regard to plant growth regulators for in vitro propagation of sweet orange cultivars from nodal segment explants. The nodal segment explants were collected from three citrus cultivars, Washington Navel, Valencia and Tangelo. Murashige and Skoog (MS) medium supplemented with sucrose and different concentrations of growth regulators was used for shoot proliferation and root induction, and the optimum medium composition was assessed. The patent for Citrus Tissue Culture was obtained from the Office of Research Affairs, Haramaya University. The results indicate that the highest shoot response was recorded for Washington's navel with maximum shoot proliferation rate (99.75%), shoot number per explant (1.76), shoot length (10.70 cm), leaf number per explants (3.54) after three weeks of culture. In all experiments, no growth was observed for the basal MS medium. Phytohormone combinations of IAA (1.2 mg/L) and kinetin (2.0 mg/L) were found to be the best for shoot proliferation. Among the cultivars, there were significant differences for the highest rooting rate (81.255), root number (2.22), and root length (2.95 cm) variables for Washington Navel. The lowest rooting rate (48.45%), root number (1.47) and root length (2.26 cm) were observed for Valencia. The highest rooting rate (84.90%), root number per microshoot (2.22) and root length (3.05 cm) was on MS medium supplemented with 1.5 mg/L NAA. A comparison of different concentrations of IAA and NAA on root induction of microshoots from nodal segments of citrus cultivars demonstrated NAA was a more effective hormone than IAA.

Optimization of Stationary Liquid Fermentation Conditions for N-Methylsansalvamide Production by the Endophytic Strain Fusarium sp. R1

To improve the yield of the therapeutic agent N-methylsansalvamide (SA), optimization of stationary liquid fermentation conditions was conducted on an endophytic strain, Fusarium sp. R1, at flask level. Using a One-Factor-At-a-Time approach, the fermentation conditions for SA production were determined as follows: fermentation time of 13 d, 24 °C, initial pH of 6.5, seed age of 24 h, inoculum size of 5.0% (v/v), loading volume of 50% (v/v), and 20.0 g/L salinity. Sucrose, tryptone, and yeast extract were found to be the best sources of carbon and nitrogen. Using response surface methodology, the optimal medium compositions consisted of 22.5 g/L sucrose, 16.5 g/L tryptone, and 0.024 g/L yeast extract. Verification tests suggested that the SA yield under these optimal conditions reached up to 536.77 ± 2.67 mg/L, which was increased by almost ten times the initial yield (54.05 ± 3.45 mg/L). The findings indicate that a high SA production yield can be achieved by stationary culture of strain R1 under proper fermentation conditions using a low-cost medium. This study paves the way toward industrial-scale SA production by strain R1 for new drug development.

Combinatorial optimization of pathway, process and media for the production of p-coumaric acid by Saccharomyces cerevisiae.

Microbial cell factories are instrumental in transitioning towards a sustainable bio-based economy, offering alternatives to conventional chemical processes. However, fulfilling their potential requires simultaneous screening for optimal media composition, process and genetic factors, acknowledging the complex interplay between the organism's genotype and its environment. This study employs statistical design of experiments to systematically explore these relationships and optimize the production of p-coumaric acid (pCA) in Saccharomyces cerevisiae. Two rounds of fractional factorial designs were used to identify factors with a significant effect on pCA production, which resulted in a 168-fold variation in pCA titre. Moreover, a significant interaction between the culture temperature and expression of ARO4 highlighted the importance of simultaneous process and strain optimization. The presented approach leverages the strengths of experimental design and statistical analysis and could be systematically applied during strain and bioprocess design efforts to unlock the full potential of microbial cell factories.

Optimization of Fermentation Process for New Anti-Inflammatory Glycosylceramide Metabolite from Aspergillus sp.

A novel ceramide compound, named Aspercerebroside A (AcA), was successfully isolated from the ethyl acetate layer of the marine symbiotic fungus Aspergillus sp. AcA exhibited notable anti-inflammatory activity by effectively inhibiting the production of nitric oxide (NO) in RAW 264.7 cells at concentrations of 30 μg/mL and 40 μg/mL, offering a promising avenue for the treatment of inflammatory diseases. To optimize the yield of glycosylceramide (AcA), a series of techniques, including single-factor experiments, orthogonal experiments, and response surface optimization, were systematically employed to fine-tune the composition of the fermentation medium. Initially, the optimal carbon source (sucrose), nitrogen source (yeast extract powder), and the most suitable medium salinity (14 ppt) were identified through single-factor experiments. Subsequently, orthogonal experiments, employing an orthogonal table for planning and analyzing multifactor experiments, were conducted. Finally, a mathematical model, established using a Box-Behnken design, comprehensively analyzed the interactions between the various factors to determine the optimal composition of the fermentation medium. According to the model's prediction, when the sucrose concentration was set at 37.47 g/L, yeast extract powder concentration at 19.66 g/L, and medium salinity at 13.31 ppt, the predicted concentration of glycosylceramide was 171.084 μg/mL. The experimental results confirmed the model's accuracy, with the actual average concentration of glycosylceramide under these conditions measured at 171.670 μg/mL, aligning closely with the predicted value.

Heavy metal tolerance, and metal biosorption by exopolysaccharides produced by bacterial strains isolated from marine hydrothermal vents

The present study highlights heavy metal tolerance, EPS production, and biosorption capacity of four hydrothermal vent bacterial strains, namely Exiguobacterium aquaticum, Mammaliicoccus sciuri, Micrococcus luteus, and Jeotgalicoccus huakuii against As, Cd, Cr, Cu, Co, Pb and Ni. The biosorption assay showed high removal efficiency of As (83%) by E. aquaticum, Cd (95%) by M. sciuri, Cu (94%) by M. luteus, and Ni (89%) by J. huakuii and their produced EPS with these metals in aqueous solution were 84%, 85%, 98%, and 91%, respectively. The maximum EPS yield was attained by optimized medium composition consisting of 1% Xylose, and 1% NaCl at pH 7. In metal-amended conditions, the four bacterial strains showed induced EPS production in the initial concentrations. SEM with EDX and CLSM images showed that the growth and EPS production of bacterial strains were affected by metal ion concentrations. A phenol sulphuric acid method and BCA assay were used to identify both the carbohydrate and total protein content of four extracted EPS. A DPPH assay revealed that EPS influences free radical scavenging and has a highly enhanced synergistic effect with its antioxidant activity. FT-IR analysis of four extracted EPS showed the shifting of peaks in the functional groups of EPS before and after adsorption of metal ions. At pH 5 and after 60 min contact time metal removal efficiency and adsorption capacity increased as calculated for As, Cd, Cu, and Ni by four extracted EPS: (86%, 20 mg/g), (74%, 19 mg/g), (94%, 60 mg/g) and (89%, 32 mg/g) and (89%, 16 mg/g), (85%, 16 mg/g), (96%, 22 mg/g) and (91%, 16 mg/g), respectively. The Langmuir compared to the Freundlich model was found to better represent the adsorption by EPS providing maximum adsorption capacities for As (34.65 mg/g), Cd (52.88 mg/g), Cu (24.91 mg/g), and Ni (58.38 mg/g).

Medium Optimization and Fermentation Kinetics for Antifungal Compounds Production by an Endophytic Paenibacillus polymyxa DS-R5 Isolated from Salvia miltiorrhiza.

An endophytic bacterium Paenibacillus polymyxa DS-R5 which can effectively inhibit the growth of pathogenic fungi was isolated from Salvia miltiorrhiza in our previous study. By using hydrochloric acid precipitation, methanol extraction, silica gel column isolation, dextran gel chromatography column, and HPLC, 3 compounds with antifungal activity were isolated. To further improve the production of antifungal compounds produced by this strain, fermentation medium was optimized using one-factor-at-a-time, Plackett-Burman design, and Box-Behnken design experiments. Through statistical optimization, the optimal medium composition was determined to be as follows: 14.7g/l sucrose, 20.0g/l soluble starch, 7.0g/l corn steep liquor, 10.0g/l(NH4)2SO4, and 0.7g/lKH2PO4. In this optimized medium, the highest titer of antifungal compounds reached 3452 U/ml, which was 123% higher than that in the initial medium. In addition, in order to guide scale-up for production, logistic and Luedeking-Piret equations were proposed to predict the cell growth and antifungal compounds production. The fermentation kinetics and empirical equations of the coefficients (X0, Xm, μm, α, and β) for the two models were reported, which will aid the design and optimization of industrial processes. The degrees of fit between calculated values of the model and the experimental data were 0.989 and 0.973, respectively. The results show that the cell growth and product synthesis models established in this study may better reflect the dynamic process of antifungal compounds production and provide a theoretical basis for further optimization and on-line monitoring of the fermentation process.

Transcriptomic profiling of adding cobalt chloride to improve dendrobine-type total alkaloid production.

Trichoderma longibrachiatum UN32 is known for its efficient production of dendrobine-type total alkaloids (DTTAs). This study aimed to determine the optimal medium composition for the UN32 strain using response surface methodology. Key factors, including glucose, beef extract, and CoCl2, were selected through the Plackett-Burman design. Subsequently, a factorial optimization approach was employed using the steepest ascent design, and 17 trial sets were completed via the Box-Behnken design. The optimal medium composition was found to consist of 29.4g/L of glucose, 17.3g/L of beef extract, and 0.28mmol/L of CoCl2. This optimized medium resulted in an impressive 80.8% increase in mycelial dry weight (reaching 12.303g/L) and a substantial 76.4% boost in DTTA production (reaching 541.63 ± 46.95μg). Furthermore, the fermentation process was scaled up to a 5-L bioreactor, leading to a DTTA production approximately 1.95 times than the control. Transcriptome analysis of strain UN32 in response to CoCl2 supplementation revealed significant changes in the expression of critical genes associated with the TCA cycle and L-valine, L-leucine, and L-isoleucine biosynthesis changed. These alterations resulted in a heightened influx of acetyl-CoA into DTTA production. Additionally, the expression of genes related to antioxidant enzymes was modified to maintain homeostasis of reactive oxygen species (ROS). A potential mechanism for the accumulation of DTTAs based on ROS as a signal transduction was proposed. These findings provide valuable insights into the regulatory mechanisms of DTTA biosynthesis, potentially offering a method to enhance the production of secondary metabolites in the UN32 strain. KEY POINTS: • After the RSM optimization, there is a substantial increase of 80.8% in biomass production and a significant 76.4% rise in DTTA production. • Transcriptome analysis revealed that the inclusion of CoCl2 supplements resulted in an enhanced influx of acetyl-CoA. • Proposed a mechanism for the accumulation of DTTAs for the role of ROS as a signal transduction pathway.

An in vitro agent-based modelling approach to optimization of culture medium for generating muscle cells.

Methodologies for culturing muscle tissue are currently lacking in terms of quality and quantity of mature cells produced. We analyse images from in vitro experiments to quantify the effects of culture media composition on mouse-derived myoblast behaviour and myotube quality. Metrics of early indicators of cell quality were defined. Images of muscle cell differentiation reveal that altering culture media significantly affects quality indicators and myoblast migratory behaviours. To study the effects of early-stage cell behaviours on mature cell quality, metrics drawn from experimental images or inferred by approximate Bayesian computation (ABC) were applied as inputs to an agent-based model (ABM) of skeletal muscle cell differentiation with quality indicator metrics as outputs. Computational modelling was used to inform further in vitro experiments to predict the optimum media composition for culturing muscle cells. Our results suggest that myonuclei production in myotubes is inversely related to early-stage nuclei fusion index and that myonuclei density and spatial distribution are correlated with residence time of fusing myoblasts, the age at which myotube-myotube fusion ends and the repulsion force between myonuclei. Culture media with 5% serum was found to produce the optimum cell quality and to make muscle cells cultured in a neuron differentiation medium viable.

Establishment of an In Vitro Micropropagation Protocol for Hibiscus moscheutos L. ‘Berry Awesome’

Hibiscus moscheutos L. ‘Berry Awesome’ is a complex hybrid of the new Proven Winners Summerific series of varieties with highly ornamental characteristics. Micropropagation of highly ornamental varieties is important for mass production of planting material for commercial purposes. The traditional methods for propagating Hibiscus varieties, such as cuttings or seed propagation, however, do not guarantee high rates of production of high-quality seedlings. To solve this problem, an attempt was made to develop protocols for micropropagation of Hibiscus moscheutos L. ‘Berry Awesome’ in vitro on agar and liquid medium using a bioreactor system, followed by ex vitro adaptation of the regenerants. The optimal method for sterilization of nodal explants as well as the optimal composition of the initiation medium for shoot proliferation and rooting were determined. For micropropagation on a liquid medium, a rocker-type bioreactor was used, and its advantages over micropropagation on an agar medium were demonstrated. The results showed that the best sterilization method for nodal segment explants was as follows: pretreatment by rinsing with running tap water, sterile water, and distilled water for 70 min and soaking for 5 min in a mixture of solutions of ethyl alcohol (96%), hydrogen peroxide (38%), and water in a ratio of 1:1:2. In this case, live and sterile explants accounted for 62.6%. The optimal initiation medium for axillary buds in nodal segments was the Murashige and Skoog (MS) medium supplemented with 0.1 mg L−1 N-(2-chloro-4-pyridinyl)-N’-phenylurea (CPPU), which resulted in 73.3% of axillary buds being induced. The optimal solid proliferation medium was MS medium supplemented with 0.1 mg L−1 CPPU with a proliferation coefficient of 5.8. In a liquid medium, the optimal concentration of CPPU was 0.05 mg L−1 with a proliferation coefficient of 9.2. The best medium for rooting/shoots with agar and in bioreactors was MS medium with the addition of 0.1 mg L−1 indole-3-butyric acid (IBA). The highest rooting rate was 99.0% in both types of media, and the survival rate of plantlets was 88.7% in solid media and 98.7% in the bioreactor.

Using Static Culture Method to Increase the Production of Acetobacter Xylinum Bacterial Cellulose

ABSTRACT Bacterial cellulose is a natural polymer, usually produced through synthetic (chemically defined) or natural media. However, the current methods of bacterial cellulose production are not ideal because of their low productivity and large number of byproducts. In order to improve the yield of bacterial cellulose, the influence of medium composition, culture time, culture temperature and culture pH on the production of bacterial cellulose by Acetobacter xylinum ATCC 23,767 was studied by single factor method. The optimal medium composition for Acetobacter xylinum can be analyzed by Thermal field emission scanning electron microscopy (TF-SEM) as glucose (20 g/L), yeast extract (5 g/L), peptone (5 g/L), citric acid (2 g/L). Using the logarithmic calculation method, we can determine that the optimal culture conditions are controlled by initial pH of the culture medium of 6 and constant static culture at 28°C for 9 days. Using TF-SEM, Fourier transform infrared (FT-IR) spectroscopy and water contact angle test, the best post-treatment solution can be selected as sodium hydroxide (1%). The results showed that the yield of bacterial cellulose could reach up to 8.5 g/L under the above conditions, and the obtained product exhibited a dense three-dimensional network structure.

Study of the polysaccharide production by the microalgae C-1509 Nannochloris sp. Naumann

Biologically active compounds, including polysaccharides isolated from microalgae, have various properties. Although Nannochloropsis spp. have the potential to produce secondary metabolites important for biotechnology, only a small part of the research on these microalgae has focused on their ability to produce polysaccharide fractions. This study aims to evaluate the physicochemical growth factors of Nannochloropsis spp. microalgae, which ensure the maximum accumulation of polysaccharides, as well as to optimize the parameters of polysaccharide extraction. The optimal nutrient medium composition was selected to maximize biomass and polysaccharide accumulation. The significance of selecting the extraction module and extraction temperature regime, as well as the cultivation conditions (temperature and active acidity value) is emphasized. Important chemical components of polysaccharides responsible for their biological activity were identified

Оптимизация состава криопротектора для сохранения репродуктивных клеток африканского сома

The article presents a study on the selection of the optimal composition of the protective medium for cryopres-ervation of the reproductive cells of male African catfish. Glycerol, dimethyl sulfoxide (DMSO) at concentrations of 3, 5 and 10% were studied as the main cryoprotective additives. The influence of these compounds on the quality indicators of catfish sperm (the percentage and time of sperm movement after activation), the level of sperm lipid peroxidation, and the activity of the antioxidant enzyme catalase before and after the cryopreservation process was assessed. A decrease in fish breeding indicators of the quality of African catfish sperm (the proportion of living cells, motility time) was established when DMSO was added to the basic cryomedium. The greatest decrease in the percentage of sperm motility before freezing is observed when DMSO is added at a concentration of 10%. A dose-dependent effect of a decrease in sperm activity after freezing was noted with an increase in the addition of DMSO to the cryogenic medium from 3 to 10%. It has been shown that 3% glycerol in a cryogenic medium does not contribute to the preservation of cells during freezing, cryodamage caused by the formation of ice crystals leads to a significant decrease in the motility of catfish sperm. The best indicators of sperm quality were obtained when using glycerol, the most effective concentration is 5% (40% of mobile reproductive cells within 53 s). The addition of 5% glycerol to the cryomedium contributes to the greatest decrease in the level of peroxidation of sperm lipids and an increase in catalase activity, which is fully consistent with the results on sperm motility. The calculated correlation coefficients confirm the dependence of the reproductive qualities of gametes on the antioxidant status of cells. The results indicate the importance of using antioxidants as cryoprotective additives to improve the quality of African catfish sperm breeding and indicate the need for more research to evaluate the effect of adding new potentially effective and safe antioxidants as cryoprotective additives.

High-level production of poly-γ-glutamic acid by a newly isolated Bacillus sp. YJY-8 and potential use in increasing the production of tomato

Strain YJY-8, a new γ-polyglutamic acid producer, was separated from fermented soybean paste samples. The strain was identified as a genus of Bacillus by morphological and 16S rDNA sequence analysis and was named Bacillus sp. YJY-8. The optimal medium composition and cultural conditions were studied using a single-factor experiment and a response surface experiment. The optimized medium consisted of monosodium glutamate 70 g/L, glucose 54.3 g/L, glycerol 31.8 g/L, ammonium sulfate 11.1 g/L, yeast extract 3.2 g/L, tryptone 1.5 g/L, L-glutamic acid 6.8 g/L, MgSO4 7H2O 0.5 g/L, FeCl3 6H2O 0.02 g/L, KH2PO4 0.9 g/L, CaCl2 0.03 g/L, MnSO4 H2O 0.3 g/L, ammonium molybdate 0.02 g/L, pH 7.0. The optimal cultivation conditions were 35 °C and pH 7.0. Under the optimized conditions, after 48 hr of cultivation, the highest shaking flask fermentation level of γ-PGA reached 65.2 ± 0.36 g/L. In addition, through fed-batch fermentation in 30 L fermenters, the fermentation level of γ-PGA reached its highest level at 88.42 g/L and productivity was 1.23 g/(L hr) after 72 hr. Then, the effect of γ-PGA on tomato yield was investigated. At the seedling stage, the plant height and stem diameter of γ-PGA treated plants increased by 5.69 and 15.735% after spraying γ-PGA for 19 days. During the flowering and fruiting period, the stem diameter of the γ-PGA treatment group increased by 6.74%, with a maximum increase of 11.65%. The number of fruit branches increased by 0.56–16.29% and the number of fruit sets increased by 1.01–28.47%. At the fruit maturation stage, the yield of tomatoes increased by 10.51, 14.27, and 5.83%.

Optimization of Mustard Pakcoy Growth: The Influence of Media Composition and Coconut Water Concentration

This research aimed to investigate the effect of different media compositions and varying coconut water concentrations on the growth and yield of mustard pakcoy (Brassica rapa var. parachinensis). The study was conducted in Modong Village, Sidoarjo Regency, with further laboratory analysis at the Faculty of Science and Technology, Universitas Muhammadiyah Sidoarjo. The experimental design followed a Randomized Block Design (RAK) in a factorial arrangement with two primary factors (media composition and coconut water concentration) and four replications. While no significant interaction was observed between the media composition and the coconut water concentration, independent effects of each factor were notable. The media composition significantly influenced the leaf count at 7, 28, and 35 days after planting (DAP) and the wet weight at 35 DAP. The optimum media composition included 2 kg of topsoil, 90 g of goat manure, and 45 g of rice husk charcoal per polybag, which resulted in the highest leaf count and wet weight. Additionally, coconut water concentration significantly affected the stem diameter at 35 DAP, with a concentration of 100 ml per liter of water yielding the maximum stem diameter. The findings imply potential growth optimization strategies for mustard pakcoy, enhancing yields via targeted alterations in growing media and the use of coconut water, which could be of international interest to agronomists, horticultural scientists, and sustainable agriculture practitioners.

Optimization of Rhodococcus erythropolis JCM3201T Nutrient Media to Improve Biomass, Lipid, and Carotenoid Yield Using Response Surface Methodology.

The oleaginous bacterium Rhodococcus erythropolis JCM3201T offers various unique enzyme capabilities, and it is a potential producer of industrially relevant compounds, such as triacylglycerol and carotenoids. To develop this strain into an efficient production platform, the characterization of the strain's nutritional requirement is necessary. In this work, we investigate its substrate adaptability. Therefore, the strain was cultivated using nine nitrogen and eight carbon sources at a carbon (16 g L-1) and nitrogen (0.16 g L-1) weight ratio of 100:1. The highest biomass accumulation (3.1 ± 0.14 g L-1) was achieved using glucose and ammonium acetate. The highest lipid yield (156.7 ± 23.0 mg g-1DCW) was achieved using glucose and yeast extract after 192 h. In order to enhance the dependent variables: biomass, lipid and carotenoid accumulation after 192 h, for the first time, a central composite design was employed to determine optimal nitrogen and carbon concentrations. Nine different concentrations were tested. The center point was tested in five biological replicates, while all other concentrations were tested in duplicates. While the highest biomass (8.00 ± 0.27 g L-1) was reached at C:N of 18.87 (11 g L-1 carbon, 0.583 g L-1 nitrogen), the highest lipid yield (100.5 ± 4.3 mg g-1DCW) was determined using a medium with 11 g L-1 of carbon and only 0.017 g L-1 of nitrogen. The highest carotenoid yield (0.021 ± 0.001 Abs454nm mg-1DCW) was achieved at a C:N of 12 (6 g L-1 carbon, 0.5 g L-1 nitrogen). The presented results provide new insights into the physiology of R. erythropolis under variable nutritional states, enabling the selection of an optimized media composition for the production of valuable oleochemicals or pigments, such as rare odd-chain fatty acids and monocyclic carotenoids.