Optimal Culture Conditions Research Articles

GC-MS Analysis of Polysaccharides from an Intergeneric Hybrid of Pleurotus florida and Cordyceps militaris: A Comparative Study.

Edible and medicinal mushrooms are valuable sources of polysaccharides, known for their dual roles as immunostimulants and immunosuppressants. This study aimed to enhance polysaccharide content by fusing two mushroom species, P. florida and C. militaris, while exploring their antioxidant and antibacterial potential. These mushrooms have diverse health benefits, including lowering high cholesterol, providing anti-inflammatory effects, supporting diabetes management, aiding in cancer treatment, and enhancing the efficacy of COVID-19 vaccines. Successful hyphal fusion was achieved, and optimal culture conditions were determined using response surface methodology. The hybrids exhibited superior growth compared to the parental strains. Hyphal fusion improved several attributes, resulting in diverse hybrids with increased biomass and metabolite production. FTIR analysis confirmed the presence of exopolysaccharides, with concentrations measured at 28.4g/L (P1), 31.50g/L (CD), and 36.74g/L (F3). GC-MS analysis identified various bioactive metabolites, including a higher concentration of dimethyl palmitamine in the hybrid, a novel compound, butanenitrile, 2-(methoxymethoxy), which was not found in the parental strains. These compounds are likely responsible for the enhanced antimicrobial and antioxidant activities.

Comprehensive cultivation of the broiler gut microbiota guides bacterial isolation from chickens.

Chicken gut microbiota plays an important role in maintaining their physiological health. However, the cultivability of chicken gut microbiota is not well understood, limiting the exploration of certain key gut bacteria in regulating intestinal health and nutritional metabolism. This study aimed to examine the cultivability of chicken cecal microbiota and to provide guidance for future chicken gut microbiota cultivation. A total of 58 different culture conditions were applied to culture broiler cecal microbiota, and the culture-dependent (CD; pooled colonies form each plate) and culture-independent (CI; broiler cecal contents) samples were collected for 16S rRNA gene sequencing and microbial analysis. The CD methods detected higher microbial richness (3,636 vs 2,331 OTUs) than CI methods, and the recovery rates of bacterial OTUs and genera reached 43.6% and 68.9%, respectively. The genera of Bacteroides (19.9%), Alistipes (11.0%) and Barnesiella (10.7%) were highly abundant detected by CI methods, however, there occupied a small proportion (<1.0%) of total cultured microbiota in CD methods. We then developed reference figures and tables showing optimal cultivation conditions for different gut bacteria taxa. Moreover, 81 different lactic acid bacteria strains covering 5 genera were isolated, and 15 strains had less than 97.0% similarity to known bacteria in the national center for biotechnology information (NCBI) online database. Overall, this study provides preliminary guidance in culturing specific gut microbiota from chickens, which will contribute to future studies to characterize the biological functions of key microbes in chicken nutritional metabolism and health.

Optimization of culturing conditions for leafcutter ant associated, antimicrobial producing Pseudonocardia by adding ants´ cuticular hydrocarbons

Optimization of culturing conditions for leafcutter ant associated, antimicrobial producing Pseudonocardia by adding ants´ cuticular hydrocarbons

Light Intensity Effects on Productivity and Post-Harvest Quality in Perilla frutescens Cultivated in CEA

Leafy vegetables, mainly lettuces, are currently the main crop cultivated in controlled environment agriculture (CEA), including vertical farming and plant factories. There is a rising demand to expand this portfolio with a wider variety of underutilized edible plants containing various bioactive compounds and sensory properties seeking to enrich human diets. However, the optimal cultivation conditions for these underutilized plants significantly differ from those optimized for lettuce, basil, and other popular CEA crops. Therefore, this study aims to explore the impacts of light-emitting diode (LED) lighting intensity (photosynthetic photon flux density, PPFD) on green leaf Perilla frutescens cultivated in CEA. Plants were grown under four levels of LED lighting PPFDs from 150 to 300 µmol m−2s−1 for 4 weeks. Plant biomass productivity, soluble sugar contents, antioxidant properties (DPPH, ABTS free radical scavenging activities, FRAP antioxidant power), and total contents of phenolic compounds in leaves were evaluated at harvesting time. Further, harvested plant material was stored in the dark, at +6 °C, and the water content, water loss and transpiration rate, leaf sugar contents, and antioxidant properties were monitored 1, 3, and 5 days after harvesting. The summarized data suggest that higher cultivation lighting PPFD results in better harvest quality preservation during post-harvest storage.

Change in p53 nuclear localization in response to extracellular matrix stiffness

AbstractChondrocytes are commonly applied in regenerative medicine and tissue engineering. Thus, the discovery of optimal culture conditions to obtain cells with good properties and behavior for transplantation is important. In addition to biochemical cues, physical and biomechanical changes can affect the proliferation and protein expression of chondrocytes. Here we investigated the effect of extracellular matrix stiffness on mouse articular chondrocyte phenotype, growth, and subcellular p53 localization. Chondrocytes were seeded on collagen‐coated substrates varying in elasticity: 0.5 and 100 kPa. Immunocytochemical staining and immunoblotting showed that a softer substrate significantly increased p53 nuclear localization in chondrocytes. Furthermore, we identified microRNA‐532 (miR‐532) as a potential p53 target gene to influence cell function, indicating a new target for tissue engineering. These findings provide insight into the influence of physical cues on cell phenotype maintenance and could help improve understanding of cartilage‐related pathologies such as osteoarthritis.

Exploring Metalloprotease from Dunaliella sp.: Production, Regulation, and Structural Insight

A green microalgal strain, identified as Dunaliella sp., was isolated from the Tunisian southern region. The enhancement of its protein and protease production was performed through culture condition optimization using the response surface methodology. The optimal conditions for protein and protease production were found to be, respectively, (i) NaCl concentrations of 135 and 45.55 g/L, (ii) NaHCO3 concentrations of 0.5 and 1.5 g/L, (iii) temperature of 28 °C for both, and (iv) light intensities of 400 and 100 µmol photons/m2/s. The optimization led to an increase in microalgae protein content from 11.98% ± 0.26 to 18.39% ± 0.10 and microalgae proteolytic activity from 7.36 ± 0.74 U/mg to 12.54 ± 0.86 U/mg. Specific focus was attributed to ATP-dependent metalloprotease, namely, FtsH2, which is involved in numerous cellular processes including cell division, cell differentiation, signal transduction, and stress response. Differential expression of the FtsH2 gene under various stress conditions showed that this expression was upregulated in response to salt stress, gibberellic acid, and Indole-3-butyric acid. A 3D modeling demonstrated two possible arrangements where the ATPase ring shows either a perfect six-fold symmetry with an open circular entrance covering the crucial pore residues, or a translocated model triggered by substrate binding inward movement of the aromatic pore residues.

Nostoxanthin Biosynthesis by Sphingomonas Species (COS14-R2): Isolation, Identification, and Optimization of Culture Conditions

Nostoxanthin, a yellow pigment, belongs to the xanthophyll group of carotenoids found in various species of bacteria and cyanobacteria. Several species of Sphingomonas can produce appropriate carotenoids for survive in various environments. This comprise nostoxanthin, a significant carotenoid. The study isolated the Sphingomonas species strain COS14-R2 from the Cosmos bipinnatus and identified it through the whole-genome sequence. The strain consists of a circular chromosome with a length of 3,677,457 base pairs.s ecThe genome consists of three carotenoid biosynthesis genes, specifically crtB (phytoene synthase), crtI (phytoene desaturase), and crtY (Lycopene beta-cyclase), which are involved in the synthesis of nostoxanthin. The strain has a circular, undulated colony morphology with a deep yellow color. It demonstrates optimal growth in liquid media at 25 to 35 °C and exhibits a high tolerance for pH levels between 5 and 11 and requires adequate quantities of carbon and nitrogen. We observed the highest concentration of nostoxanthin was recorded at 35 °C, pH of 7.5, glucose concentration of 40 g L−1, and a yeast extract concentration of 5 g L−1 during dark incubation. The fed-batch fermentation process produced nostoxanthin at a concentration of 217.22 ± 9.60 mg L−1, with a selectivity of 72.32% and a productivity of 2.59 g/L/h. The dry biomass extract was purified using column chromatography. The LC–MS/MS analysis of the purified fraction indicated that the molecular weight of nostoxanthin is 600.5098 m/z. The DPPH assay result of 75.5 ± 0.33% indicates nostoxanthin is highly effective in scavenging free radicals.

Navigating stem cell culture: insights, techniques, challenges, and prospects.

Stem cell research holds huge promise for regenerative medicine and disease modeling, making the understanding and optimization of stem cell culture a critical aspect of advancing these therapeutic applications. This comprehensive review provides an in-depth overview of stem cell culture, including general information, contemporary techniques, encountered problems, and future perspectives. The article begins by explaining the fundamental characteristics of various stem cell types, elucidating the importance of proper culture conditions in maintaining pluripotency or lineage commitment. A detailed exploration of established culture techniques sheds light on the evolving landscape of stem cell culture methodologies. Common challenges such as genetic stability, heterogeneity, and differentiation efficiency are thoroughly discussed, with insights into cutting-edge strategies and technologies aimed at addressing these hurdles. Moreover, the article delves into the impact of substrate materials, culture media components, and biophysical cues on stem cell behavior, emphasizing the intricate interplay between the microenvironment and cell fate decisions. As stem cell research advances, ethical considerations and regulatory frameworks become increasingly important, prompting a critical examination of these aspects in the context of culture practices. Lastly, the article explores emerging perspectives, including the integration of artificial intelligence and machine learning in optimizing culture conditions, and the potential applications of stem cell-derived products in personalized medicine. This comprehensive overview aims to serve as a valuable resource for researchers and clinicians, fostering a deeper understanding of stem cell culture and its key role in advancing regenerative medicine and biomedical research.

In vitro strategy to enhance the production of bioactive polyphenols and caffeoylputrescine in the hairy roots of Physalis peruviana L.

The Rhizobium rhizogene-transformed root culture from Physalis peruviana L. (P. peruviana) may be a promising and novel source of valuable phenolics, including caffeoylputrescine (CP), which is known for antioxidant, antidiabetic, insect-resistant, disease-resistant, and neuroprotective properties. In this study, to improve the production efficiency of phytochemical components in P. peruviana hairy root cultures, we optimized various culture conditions, including the inoculum size, liquid volume, culture media type, carbon source, sucrose concentration, initial pH, and application of elicitors, to enhance the total phenolic content and CP yield in these hairy root cultures. The findings indicate that the use of sucrose as carbon source resulted in the highest biomass (13.28 g DW/L), total phenolic content (6.26 mg/g), and CP yield (2.40 mg/L). The White medium excelled in enhancing the total phenolic content (9.35 mg/g), whereas the B5 medium was most effective for the biomass (13.38 g DW/L) and CP yield (6.30 mg/L). A sucrose concentration of 5% was best for the biomass (18.40 g DW/L), whereas a sucrose concentration of 4% was ideal for the CP yield. Optimal culture conditions were as follows: an inoculum size of 0.5 g/100 mL, a liquid volume of 100 mL in a 250-mL flask, B5 medium, 4% sucrose, and a pH of 5.5. Among the tested elicitors, methyl jasmonate (MeJA) at 100 µM significantly increased the biomass (21.3 g/L), total phenolic content (23.34 mg/g), and CP yield (141.10 mg/L), which represent 0.96-, 2.12-, and 13.04-fold increases, respectively, over the control after 8 days. The optimized HR culture of P. peruviana provides a promising system to enhance the production of CP for pharmaceutical applications.

Biomass Accumulation, Contaminant Removal, and Settling Performance of Chlorella sp. in Unsterilized and Diluted Anaerobic Digestion Effluent

Microalgae demonstrate significant efficacy in wastewater treatment. Anaerobic digestion effluent (ADE) is regarded as an underutilized resource, abundant in carbon, nitrogen, phosphorus, and other nutrients; however, the presence of inhibitory factors restricts microalgal growth, thereby preventing its direct treatment via microalgae. The purpose of this study was to dilute ADE using various dilution media and subsequently cultivate Chlorella sp. to identify optimal culture conditions that enhance microalgal biomass and water quality. The effects of various dilution conditions were assessed by evaluating the biomass, sedimentation properties, and nutrient removal efficiencies of microalgae. The results demonstrate that microalgal biomass increases as the dilution ratio increased. The microalgae biomass in the treatments diluted with simulated wastewater was significantly higher than that with deionized water, but their effluent quality failed to meet discharge standards. The treatment diluted with deionized water for 10 times exhibited abundant microbial biomass with strong antioxidant properties. The corresponding total phosphorus concentration in the effluent (6.96 mg/L) adhered to emission limits under the Livestock and Poultry Industry Pollutant Emission Standards (8 mg/L), while ammonia nitrogen concentration (90 mg/L) was near compliance (80 mg/L). The corresponding microbial biomass, with a sludge volume index (SVI30) of 72.72 mL/g, can be recovered economically and efficiently by simple precipitation. Its high protein (52.07%) and carbohydrate (27.05%) content, coupled with low ash (10.75%), makes it a promising candidate for animal feed and fermentation. This study will aid in understanding microalgal growth in unsterilized ADE and establish a theoretical foundation for cost-effective ADE purification and microalgal biomass production.

Improved exopolysaccharide production by Lactiplantibacillus plantarum Z-1 under hydrogen peroxide stress and its physicochemical properties

In this study, a strain with good exopolysaccharide (EPS)-producing ability was isolated from the fermented Benincasa hispida and identified as Lactiplantibacillus plantarum Z-1. Its EPS production was further improved by H2O2 stress under optimized culture conditions, increasing from 180 ± 0.45 mg/L to 409.52 ± 2.16 mg/L. Purification of EPS with DEAE-52 and subsequent Sephadex G-100 column chromatography provided three fractions, namely, EPS-0, EPS-1 and EPS-3, respectively. The molecular weight of EPS, EPS-0, EPS-1 and EPS-3 were 85.4, 25.7, 131.88 and 93.2 kDa, respectively. EPS, EPS-1 and EPS-3 were mainly composed of glucose, rhamnose, arabinose and galactose with molar ratios of 1:0.544:0.211:0.281, 1:1.279:0.807:0.704, and 1:1.459:0.759:0.75, respectively, along with small proportions of fucose, mannose and xylose. EPS-0 was composed of glucose, arabinose, galactose and xylose, with molar ratios of 1:0.618:0.206:0.275. The structural analysis indicated that EPS-3 was mainly consisted of (1,2,4)-β-Rhap, (1,2,3)-β-Araf, (1,4)-β-Galp, T-α-Glcp units. The three purified fractions showed typical characteristics of non-Newtonian fluids and good viscoelasticity. Congo red test revealed that irregular triple-helical conformation existed in EPS and EPS-3. These physicochemical properties of EPSs make them a potential candidate for the use as a health-beneficial food additive in the food processing industry.

Biological Properties of S. warneri KYS-164 Isolated from Kefir Grains.

Staphylococcus worderi KYS-164, isolated from homemade Tibetan kefir grains, produces bacteriocin-like inhibitory substances (BLIS), which are peptides with antimicrobial properties, but have not been fully characterized. The research on BLIS will lay the foundation for mining new bacteriocins. In this study, the optimal culture conditions for the production of highly active BLIS were found to be incubation at 30 °C and 120 rpm, and the most effective extraction method was ammonium sulfate precipitation (ASP) using ammonium sulfate at 80% saturation. The postantibiotic effect (PAE) of BLIS on Staphylococcus aureus CICC 10384 is significant, with a 4 × MIC BLIS concentration able to prolong the PAE to 2.39 h. BLIS has excellent biosafety, with no deleterious effects observed at 8 × MIC concentration. Gas chromatography-ion mobility spectrometry (GC-IMS) was used to analyze the volatile compounds synthesized by Staphylococcus warneri KYS-164 during its growth. Hydroxycitronellal, ethyl pyruvate, and α-pinene were found to be unique substances produced by this strain, which can provide fresh, refreshing floral and fruity aromas as well as strong pine and resinous aromas in the process of kefir grain fermentation of milk. Analysis of the S. warneri KYS-164 genome provided insights into the major metabolic pathways in which genes expressed in this strain are involved. This study represents the first isolation of S. warneri KYS-164 from kefir grains prepared by Tibetan families, and provides a comprehensive analysis of its physicochemical properties. This research provides a solid foundation for better understanding and utilization of S. warneri KYS-164.

Design of an Innovative Method for Measuring the Contractile Behavior of Engineered Tissues.

Hypertrophic scarring is a common complication in severely burned patients who undergo autologous skin grafting. Meshed skin grafts tend to contract during wound healing, increasing the risk of pathological scarring. Although various technologies have been used to study cellular contraction, current methods for measuring contractile forces at the tissue level are limited and do not replicate the complexity of native tissues. Self-assembled skin substitutes (SASSs) were developed at the "Centre de recherche en organogénèse expérimentale de l'Université Laval/LOEX" and are used as permanent full-thickness skin grafts. The autologous skin substitutes are produced using the self-assembly method, allowing the cultured cells to produce their extracellular matrix leading to a tissue-engineered substitute resembling the native skin. The level of contraction of the SASSs during the fabrication process is patient-dependent. Thus, because of its architecture and composition, SASS is an interesting model to study skin contraction in vitro. Unfortunately, standard measurement methods are unsuited for SASS contraction assessment, mainly due to incompatibilities between the SASS manufacturing process and the current contraction force measurement methods. Here, we present an innovative contraction measurement method specifically designed to quantify the contractile behavior of tissue-engineered substitutes, without disrupting the protocol of production. The method uses C-shape anchoring frames that close at different speeds and magnitudes according to the tissue contractile behavior. A finite element analysis model is then used to associate the frame deformation to a contractile force amplitude. This article shows that the method can be used to measure the contraction force of tissues produced with cells displaying different contractile properties, such as primary skin fibroblasts and myofibroblasts. It can also be used to study the effects of cell culture conditions on tissue contraction, such as serum concentration. This protocol can be easily and affordably applied and tuned to many regenerative medicine applications or contraction-related pathological studies. Impact Statement The protocol presented in this article is a new and simple method to quantify contraction forces present in tissue-engineered substitutes. Using finite element analysis, it allows for the measurement of a contraction force rather than a surface reduction as usually provided by other tissue contraction measurement methods. The results shown are in correlation with the current literature relevant to tissue contraction. It can be easily implemented, and hence, this method will open up new avenues to study tissue contraction of living substitutes engineered with various cell types and to optimize culture conditions.

A New Enzyme for Biodiesel Production and Food Applications: Lipase of Bacillus megateriumF25 Isolated From an Aquatic Insect Rhantus suturalis

ABSTRACTThis study aimed to isolate, purify, and characterize a lipase from the gut symbiont Bacillus megaterium F25 (GenBank accession: MF597792) of the aquatic insect Rhantus suturalis, with a focus on its potential applications in biodiesel and food industries. Under optimized culture conditions, B. megaterium F25 could produce 583 U/L of lipase in shaking flask culture. The purified lipase (PL) exhibited a specific activity with 113.89 U/mg, and its molecular weight was determined as 34 kDa. The activity of PL was enhanced by methanol, ethanol, Tween‐80, Triton X‐100, Ca2+, and Mg2+, while β‐mercaptoethanol, EDTA, SDS, Fe2+, Mn2+, and Cu2+ were inhibitory. PL showed optimal activity and stability at neutral and slightly acidic pHs, as well as in a temperature range of 20°C–30°C. PL displayed strong hydrolytic activity toward plant oils and animal fats, indicating its potency for both the food industry and the remediation of oil‐contaminated environments. When tested as a catalyst, PL provided biodiesel production with a transesterification yield of 86.8% under optimized conditions (36 h reaction time, 4 mL enzyme solution, 30°C, pH 7.0, and waste cooking oil:methanol ratio of 10 mL/40 mL). This is the first report on the lipase‐producing potential of gut microbial symbionts of aquatic insects. Furthermore, B. megaterium lipase was tested for the first time as a biocatalyst for biodiesel production.

Ultraviolet-diethyl sulfate composite mutagenesis of Rhodosporidium toruloides and culture optimization to improve carotenoid production

Rhodosporidium toruloides is a carotenoid-producing oleaginous yeast, and its carotenoids are of considerable value to food, chemical, and medical industries. In this study, to improve carotenoid production capacity, the parent R. toruloides strain (CK) was compound mutagenized using ultraviolet-diethyl sulfate (UV-DES), and cultivation conditions were simultaneously optimized to enhance the carotenoid synthesis ability of the mutant strain (M5). The carotenoid content was significantly increased following UV-DES compound mutagenesis (CK = 249.34 μg/g, M5 = 535.28 μg/g). The carotenoid content of M5 was increased to 763.85 μg/g under optimal culture conditions. In addition, mutagenesis altered the carotenoid components, and torulene and torularhodin levels were also increased following mutation, leading to changes in antioxidant capacity. Real-time quantitative PCR results showed that expression of six genes (CAR1, CAR2, CrtE, CrtI, ME1, HMG1) related to carotenoid accumulation were significantly upregulated by UV-DES composite mutagenesis. In conclusion, combined UV-DES mutagenesis and optimal culture conditions promoted the biosynthesis of carotenoids in R. toruloides.

Optimization of fermentation conditions for physcion production of Aspergillus chevalieri BYST01 by response surface methodology.

This study aimed to optimize the culture conditions of the termite-derived fungus Aspergillus chevalieri BYST01 for the production of physcion, a characteristic component of the traditional herb rhubarb, which has been commercially approved as a botanical fungicide in China. First, potato dextrose broth was screened as the suitable basal medium for further optimization, with an initial yield of 28.0 mg/L. Then, the suitable carbon source, fermentation time, temperature, pH value, and the rotary shaker speed for physcion production were determined using the one-variable-at-a-time method. Based on the results of single factors experiments, the variables with statistically significant effects on physcion production were further confirmed using the Plackett-Burman design (PBD). Among the five variables, temperature, initial pH, and rotary shaker speed were identified as significant factors (P < 0.05) for physcion productivity in the PDB and were further analyzed by response surface methodology (RSM). Finally, we found that the maximum physcion production (82.0 mg/L) was achieved under the following optimized conditions:initial pH 6.6, rotary shaker speed of 177 rpm, temperature of 28 °C, and glucose concentration of 30 g/L in PDB medium after 11 d of fermentation. The yield of physcion under the optimized culture conditions was approximately threefold higher than that obtained using the basal culture medium. Furthermore, the optimum fermentation conditions in the 5-L bioreactor achieved a maximal physcion yield of 85.2 mg/L within 8 d of fermentation. Hence, response surface methodology proved to be a powerful tool for optimizing physcion production by A. chevalieri BYST01. This study may be helpful in promoting the application of physcion produced by A. chevalieri BYST01 to manage plant diseases.

Effects of Temperature and Salinity on the Growth, Reproduction, and Carotenoid Accumulation in Artemia sinica and Transcriptome Analysis

Brine shrimp (Artemia), rich in carotenoids, are widely used in intensive aquaculture to supplement nutrients and enhance the coloration of farmed organisms. This study investigates the growth, reproduction, and carotenoid accumulation in Artemia sinica under varying salinity and temperature conditions. The results showed that temperature and salinity displayed significant interactions with survival, body length, and carotenoid accumulation in the body. The optimal survival and growth conditions of A. sinica (Bohai Sea Gulf) were a temperature range of 25–30 °C and a salinity range of 30–50‰. High temperatures accelerated growth and sexual maturity at the expense of survival rates, while temperatures below 20 °C prevented ovigerous development. Extreme salinity levels negatively affected survival and growth, though high salinity promoted sexual maturity. Carotenoids in A. sinica mainly accumulate as echinenone and canthaxanthin form. Carotenoid accumulation decreased with increased temperature and salinity, and the temperature effect decreased with rising salinity. A. sinica cultivated at a salinity of 10‰ and a temperature of 25 °C exhibits the highest carotenoid content. Transcriptomic analysis revealed that high temperatures primarily affected genes related to stress response and metabolism, while high-salinity regulated genes associated with ion balance and signaling pathways. These findings provide a theoretical basis for enhancing Artemia sinica aquaculture and optimizing cultivation conditions, offering novel insights into nutritional and environmental impacts on brine shrimp biology.

Recent Advancements in Production and Extraction Methods of Phycobiliprotein C-phycocyanin by Arthrospira (Spirulina) platensis: A Mini Review.

Arthrospira platensis has been utilized as a food source since ancient times due to its rich nutrient profile. In recent years, its popularity as a dietary supplement has soared, especially due to the presence of a water-soluble phycobiliprotein, C-phycocyanin C (C-PC), which is abundant and notable for its fluorescent properties. C-PC contains the chromophore phycocyanobilin B (PCB-B), a tetrapyrrole molecule, that is why it plays a dual role as a food colorant and as nutraceutical. However, comprehensive studies have mostly evaluated C-PC's broader health-promoting properties, particularly its antioxidative and anti-inflammatory effects, which are linked to its ability to contrast oxidative stress and related pathological conditions. That is why this review explores recent advancements in optimizing culture conditions to enhance C-PC and PCB-B production, with a particular emphasis on novel extraction and purification techniques that increase yield and bioactivity. This focus on efficient production methods is crucial for expanding the commercial and therapeutic applications of C-PC, contributing to its growing relevance in the food and pharmaceutical industries.

Engineering Escherichia coli via introduction of the isopentenol utilization pathway to effectively produce geranyllinalool

BackgroundGeranyllinalool, a natural diterpenoid found in plants, has a floral and woody aroma, making it valuable in flavors and fragrances. Currently, its synthesis primarily depends on chemical methods, which are environmentally harmful and economically unsustainable. Microbial synthesis through metabolic engineering has shown potential for producing geranyllinalool. However, achieving efficient synthesis remains challenging owing to the limited availability of terpenoid precursors in microorganisms. Thus, an artificial isopentenol utilization pathway (IUP) was constructed and introduced in Escherichia coli to enhance precursor availability and further improve terpenoid synthesis.ResultsWe first constructed an artificial IUP in E. coli to enhance the supply of precursor geranylgeranyl diphosphate (GGPP) and then screened geranyllinalool synthases from plants to achieve efficient synthesis of geranyllinalool (274.78 ± 2.48 mg/L). To further improve geranyllinalool synthesis, we optimized various cultivation factors, including carbon source, IPTG concentration, and prenol addition and obtained 447.51 ± 6.92 mg/L of geranyllinalool after 72 h of shaken flask fermentation. Moreover, a scaled-up production in a 5-L fermenter was investigated to give 2.06 g/L of geranyllinalool through fed-batch fermentation. To the best of our knowledge, this is the highest reported titer so far.ConclusionsEfficient synthesis of geranyllinalool in E. coli can be achieved through a two-step pathway and optimization of culture conditions. The findings of this study provide valuable insights into the production of other terpenoids in E. coli.

High-throughput optimization of organic carbon provision strategies enables enhanced arachidonic acid production in novel microalgae

BackgroundMicroalgae are potential sustainable resources for the production of value-added chemicals that can be used as biofuels, pharmaceuticals, and nutritional supplements. Arachidonic acid (ARA), a omega-6 fatty acid, plays a crucial role in infant development and immune response, and can be used in cosmetics and pharmaceuticals. Demand for industrial-scale ARA production is continuously increasing because of its broad applicability. To address this demand, there has been a significant shift towards microorganism-based ARA production. To accelerate large-scale ARA production, it is crucial to select suitable strains and establish optimal culture conditions.ResultsHere, we isolated a novel microalga Lobosphaera incisa CFRC-1, a valuable strain that holds promise as a feedstock for ARA production. Optimal cultivation conditions were investigated using a high-throughput screening method to enhance ARA production in this novel strain. Out of 71 candidates, four organic carbon substrates were identified that could be utilized by L. incisa CFRC-1. Through flask-scale verification, fructose was confirmed as the optimal organic carbon substrate for promoting microalgal growth, total lipid accumulation, and ARA production. Subsequently, we investigated appropriate substrate concentration and cultivation temperature, confirming that the optimal conditions were 30 g L− 1 of fructose and 27 ℃ of temperature. Under these optimized conditions, biomass and ARA production reached 13.05 ± 0.40 g L− 1 and 97.98 ± 7.33 mg L− 1, respectively, representing 9.6-fold and 5.3-fold increases compared to the conditions before optimization conditions. These results achieved the highest biomass and ARA production in flask-scale cultivation, indicating that our approach effectively improved both production titer and productivity.ConclusionsThis study presents a novel microalgae and optimized conditions for enhancing biomass and ARA production, suggesting that this approach is a practical way to accelerate the production of valuable microalgae-based chemicals. These findings provide a basis for large-scale production of ARA-utilizing microalgae for industrial applications.