Growth Medium Research Articles

A user-friendly fluorescent biosensor for precise lactate detection and quantification in vitro.

As a critical metabolite, the standardization of lactate quantification is increasingly crucial. Therefore, we developed LaconicSF, a lactate-responsive biosensor exhibiting exceptional specificity in lactate detection. LaconicSF enables efficient lactate quantification in CHO cell culture medium and holds potential as a user-friendly detection tool for lactate quantification in vitro.

Numerical simulation of fluid flow characteristics and ultrasonic cavitation performance in novel-designed stirred tank sonoreactor

A novel-designed stirred tank sonoreactor was developed to well solve large-scale dispersion of nanoparticles in liquid phase system. The fluid flow characteristics and ultrasonic cavitation performance in the sonoreactor were numerically simulated by CFD method. By comparing the pressure, cavitation bubble and velocity distribution under different situations, it is found that larger ultrasonic amplitude, relatively smaller ultrasonic frequency, higher saturated vapor pressure and lower viscosity of liquid medium are beneficial to ultrasonic cavitation. Besides, the acoustic flow action is strengthened with the increase of ultrasonic frequency and decrease of gas-liquid mixture's average density. For the designed sonoreactor, it is critical that high-frequency transducers should be determined near the bottom and upper region of the kettle, and large-amplitude transducers can be confirmed in the middle position. The research findings will provide theoretical and technical supports for developing state-of-the-art sonoreactors and optimizing ultrasonic process parameters in the industrialized preparation of nanocomposites.

Effects of Inosine-5′-monophosphate Dehydrogenase (IMPDH/GuaB) Inhibitors on Borrelia burgdorferi Growth in Standard and Modified Culture Conditions

Borrelia burgdorferi’s inosine-5′-monophosphate dehydrogenase (IMPDH, GuaB encoded by the guaB gene) is a potential therapeutic target. GuaB is necessary for B. burgdorferi replication in mammalian hosts but not in standard laboratory culture conditions. Therefore, we cannot test novel GuaB inhibitors against B. burgdorferi without utilizing mammalian infection models. This study aimed to evaluate modifications to a standard growth medium that may mimic mammalian conditions and induce the requirement of GuaB usage for replication. The effects of two GuaB inhibitors (mycophenolic acid, 6-chloropurine riboside at 125 μM and 250 μM) were assessed against B. burgdorferi (guaB+) grown in standard Barbour–Stoenner–Kelly-II (BSK-II) medium (6% rabbit serum) and BSK-II modified to 60% concentration rabbit serum (BSK-II/60% serum). BSK-II directly supplemented with adenine, hypoxanthine, and nicotinamide (75 μM each, BSK-II/AHN) was also considered as a comparison group. In standard BSK-II, neither mycophenolic acid nor 6-chloropurine riboside affected B. burgdorferi growth. Based on an ANOVA, a dose-dependent increase in drug effects was observed in the modified growth conditions (F = 4.471, p = 0.001). Considering higher drug concentrations at exponential growth, mycophenolic acid at 250 μM reduced spirochete replication by 48% in BSK-II/60% serum and by 50% in BSK-II/AHN (p < 0.001 each). 6-chloropurine riboside was more effective in both mediums than mycophenolic acid, reducing replication by 64% in BSK-II/60% serum and 65% in BSK-II/AHN (p < 0.001 each). These results demonstrate that modifying BSK-II medium with physiologically relevant levels of mammalian serum supports replication and induces the effects of GuaB inhibitors. This represents the first use of GuaB inhibitors against Borrelia burgdorferi, building on tests against purified B. burgdorferi GuaB. The strong effects of 6-chloropurine riboside indicate that B. burgdorferi can salvage and phosphorylate these purine derivative analogs. Therefore, this type of molecule may be considered for future drug development. Optimization of this culture system will allow for better assessment of novel Borrelia-specific GuaB inhibitor molecules for Lyme disease interventions. The use of GuaB inhibitors as broadcast sprays or feed baits should also be evaluated to reduce spirochete load in competent reservoir hosts.

Fe and Mn Removal from Acid Mine Drainage by Utilizing Chlorella sorokiniana and Monoraphidium neglectum as Biosorbent

The mining industry generates acid mine drainage (AMD) characterized with a low pH value and high dissolved metal concentration that leads to the negative impacts on the environment and human health. The objectives of this research were to investigate the growth response of mixed culture of microalgae Chlorella sorokiniana and Monoraphidium neglectum in a liquid media contaminated with AMD; generate the optimum environmental conditions (pH value and contact time) to determine the efficiency biosorption of iron and manganese contained in the solution of AMD into the consortium of microalgae; and quantify the maximum removal amount of iron and manganese contained in the solution of AMD by utilizing microalgae consortium of Chlorella sorokiniana and Monoraphidium neglectum as biosorbent. AMD used in this research was characterized with a pH value of 1.65 with iron and manganese concentrations of 8.28 mg/L and 4.57 mg/L. The research of biosorption was conducted in 150 rpm with pH level variations of 4, 5, and 6, and contact time variations of 60, 120, and 180 min. The maximum value of iron and manganese removals occurred when pH level reached 5 at 180 min of contact time with removal efficiency of 89.73% for iron and 94.53% for manganese. The results proved that the mixed culture of microalgae namely Chlorella sorokiniana and Monoraphidium neglectum can be utilized to remove iron and manganese contained in acid mine drainage.

A high-throughput protocol for testing heat-stress tolerance in pollen

AbstractViable pollen is crucial for fertilization, but pollen is generally highly susceptible to heat stress. A quick, reliable method for testing the heat-stress tolerance of pollen is needed to improve the heat-stress tolerance in plants, but current methods require considerable space and labor. In addition, many such methods only test tolerance to a single constant temperature, making it time-consuming to screen heat tolerance over a wide temperature range and to examine the dynamics of pollen viability at different temperatures. To address this issue, we aimed to: (1) develop an easy, reliable method for measuring pollen viability at different temperatures; and (2) identify the best temperature range for screening pollen with high heat-stress tolerance. We harvested mature pollen from wheat (Triticum aestivum) plants and transferred it to a 96-well plate filled with liquid medium containing sucrose. We placed the plate in a PCR machine operating under a gradient PCR program to simultaneously test a range of temperatures. After incubating the pollen for 4 h, at temperatures ranging from 21.9 to 47 °C, we examined the pollen grains under a light microscope and employed a specific image analysis pipeline to assess the effects of temperature on pollen morphology, germination, and tube growth. This method facilitated the high-throughput screening of many pollen samples, enabling rapid, reliable, and precise analysis of pollen viability in response to temperature. Our approach should be applicable to other plant species and could be used to identify quantitative trait loci or genes influencing heat stress tolerance in pollen for breeding programs.

Educators’ experiences of using mobile learning as a professional development and knowledge retention tool during the Covid-19 pandemic

This qualitative study intentionally gathered insights from 12 educators of varying seniority across three schools in the Nkangala Education District, Mpumalanga Province, South Africa, regarding their experiences with mobile learning (facilitated through Microsoft Teams and WhatsApp Messenger) as a professional development and knowledge retention tool during the Covid-19 pandemic. The findings, derived from semi-structured interviews and document analysis, indicated that young educators positively embraced the implementation of Microsoft Teams and WhatsApp Messenger. Senior educators deemed WhatsApp Messenger less intimidating than Microsoft Teams, thereby endorsing it as the preferred medium for professional growth and knowledge retention. This led to a lack of incentive among senior educators to participate in the professional learning events facilitated by Microsoft Teams. Thus, they depended on the supplemental learning information disseminated via WhatsApp Messenger as the principal method for professional development and knowledge retention activities. The research identified multiple obstacles to utilising mobile learning for professional development and knowledge retention, specifically: insufficient ICT skills among senior educators, ineffective content facilitation by subject advisors, and inadequate communication and scheduling of professional development programs. The study advocated for a systematic coordination of: 1) a series of ICT skills training for underqualified educators; 2) peer teaching among novice, mid-career, and senior educators to address internal skills deficiencies; 3) effective communication among the district, circuits, and schools; and 4) skills audits for subject advisors to enhance competency in areas of deficiency. By presenting these recommendations, the report alerts schools, circuits, and districts to the likelihood that the Covid-19-induced adoption of hybrid professional development will become the standard practice.

Respiratory Delivery of Lacticaseibacillus rhamnosus GG by Vibrating-Mesh and Jet Nebulisation

Background: The use of probiotic bacteria to improve lung health has been gaining interest. Although the oral delivery of probiotics and their effects are well documented, there is currently limited knowledge on the respiratory delivery of probiotics. Objectives: This study aimed to investigate whether nebulisation is suitable for delivering Lacticaseibacillus rhamnosus GG (LGG) into the lungs for the potential treatment of bacterial pulmonary infections. Methods: It compared the dose output and aerosol performance of a vibrating-mesh nebuliser (VMN) and a jet nebuliser (JN) in nebulising LGG suspended in de Man Rogosa Sharpe (MRS) broth, phosphate-buffered saline (PBS), or normal saline (0.9% w/v sodium chloride in water). Results: The VMN consistently produced a higher output than the JN for all liquid media, indicating that VMN was more efficient. The fine-particle fractions of both nebulisers were comparable for a given medium. The highest fine-particle fraction was achieved with LGG suspended in MRS broth for both nebulisers (20.5 ± 2.8% for VMN; 18.7 ± 3.4% for JN). This suggests that the aerosol performance of nebulised probiotics may depend on the medium in which the probiotic bacteria were suspended. Conclusions: Therefore, this study demonstrated that the nebulisation efficiency of LGG depended on the nebuliser type and liquid medium of the probiotic suspension.

Synergistic Role of Amino Acids in Enhancing mTOR Activation Through Lysosome Positioning.

Lysosome positioning, or lysosome cellular distribution, is critical for lysosomal functions in response to both extracellular and intracellular cues. Amino acids, as essential nutrients, have been shown to promote lysosome movement toward the cell periphery. Peripheral lysosomes are involved in processes such as lysosomal exocytosis, cell migration, and metabolic signaling-functions that are particularly important for cancer cell motility and growth. However, the specific types of amino acids that regulate lysosome positioning, their underlying mechanisms, and their connection to amino acid-regulated metabolic signaling remain poorly understood. In this study, we developed a high-content imaging system for unbiased, quantitative analysis of lysosome positioning. We examined the 15 amino acids present in cell culture media and found that 10 promoted lysosome redistribution toward the cell periphery to varying extents, with aromatic amino acids showing the strongest effect. This redistribution was mediated by promoting outward transport through SLC38A9-BORC-kinesin 1/3 axis and simultaneously reducing inward transport via inhibiting the recruitment of Rab7 and JIP4 onto lysosomes. When examining the effects of amino acids on mTOR activation-a central regulator of cell metabolism-we found that the amino acids most strongly promoting lysosome dispersal, such as phenylalanine, did not activate mTOR on their own. However, combining phenylalanine with arginine, which activates mTOR without affecting lysosome positioning, synergistically enhanced mTOR activity. This synergy was lost when lysosomes failed to localize to the cell periphery, as observed in kinesin 1/3 knockout (KO) cells. Furthermore, breast cancer cells exhibited heightened sensitivity to phenylalanine-induced lysosome dispersal compared to noncancerous breast cells. Inhibition of LAT1, the amino acid transporter responsible for phenylalanine uptake, reduced peripheral lysosomes and impaired cancer cell migration and proliferation, highlighting the importance of lysosome positioning in these coordinated cellular activities. In summary, amino acid-regulated lysosome positioning and mTOR signaling depend on distinct sets of amino acids. Combining lysosome-dispersing amino acids with mTOR-activating amino acids synergistically enhances mTOR activation, which may be particularly relevant in cancer cells.

Continuous wave laser ablation for tailored titanium nanoparticle synthesis: temperature and liquid medium effects

We reported for the first time the generation of titanium (Ti) nanoparticles (NPs) in different liquids (deionized (DI) water and sodium dodecyl sulfate (SDS) solution) and at a range of temperatures (5 °C, 28 °C, 60 °C, and 80 °C) using continuous-wave high-power laser ablation in liquid (CWLAL). The CWLAL technique is a convenient and cost-effective way to generate NPs. The key outcomes of our investigation are the effects of temperature and the liquid mediums on the average size, generation rate, shape, surface charges, and crystallographic structure of the NPs. Generated NPs show consistent spherical shape regardless of liquid medium changes and temperature variation. SDS solution notably impacts NP size and generation rate with higher surface charges than DI water. For instance, at temperatures of 28 °C and 80 °C, the generation rates in SDS solution are 316 mg/hr and 309 mg/hr, with average NP sizes of 33 nm and 34 nm, respectively. In contrast, the generation rates in DI water are 96 mg/hr and 302 mg/hr, with NP sizes of 13 nm and 20 nm, respectively. The weaker crystallographic structure observed in NPs generated in SDS solution, compared to the more robust crystallographic structure of NPs synthesized in DI water. Liquid temperature plays a significant role in determining surface charges, average particle size, and molecular structure of NPs. The choice of the liquid medium and temperature can be critical for tailoring NP characteristics to specific applications. Ongoing work is being conducted to explore the possibilities of further progress in this area to generate efficient and customized NPs.

Impact of Protein Coronas on Lipid Nanoparticle Uptake and Endocytic Pathways in Cells.

Lipid nanoparticles (LNPs), widely used in disease diagnosis and drug delivery, face the challenge of being surrounded by biological macromolecules such as proteins upon entering the human body. These molecules compete for binding sites on the nanoparticle surfaces, forming a protein corona. The impact of different types of protein coronas on LNP delivery remains unclear. In this study, we employed a newly developed, highly sensitive LNP labeling platform and analyzed the endocytosis of HeLa cells under different nutritional conditions using proteomics to address this critical issue. Our research found that under conditions of complete medium and amino acid starvation, most DNA-FITC vesicles in HeLa cells were located in the perinuclear region 4 h after transfection. In contrast, under serum starvation conditions, only a small portion of DNA-FITC vesicles were in the perinuclear region. On the other hand, through proteomics, we discovered that cells that were enriched in amino acids and complete medium contained more proteins, whereas those under serum starvation had relatively fewer enriched proteins. Through KEGG pathway enrichment analysis, we identified the phagosome and endocytosis pathways as particularly important. Lastly, differential analysis of proteins in these pathways revealed that proteins such as F-actin, Coronin, vATPase, TUBA, TUBB, MHCII, and TSP may have significant impacts on cellular endocytosis. Our research findings indicate that it is necessary to regulate cellular endocytosis based on different protein coronas to achieve optimal cytoplasmic release.

Analysis of low-density lipoprotein receptor gene mutations in a family with familial hypercholesterolemia.

Familial hypercholesterolemia (FH) is a common monogenic autosomal dominant disorder, primarily mainly caused by pathogenic mutations in the low-density lipoprotein receptor (LDLR) gene. Through phenotypic-genetic linkage analysis, two LDLR pathogenic mutations were identified in FH families: c.G1027A (p.Gly343Ser) and c.G1879A (p.Ala627Thr). Whole exome sequencing was conducted on the proband with familial hypercholesterolemia to identify the target gene and screen for potential pathogenic mutations. The suspicious responsible mutation sites in 14 family members were analyzed using Sanger sequencing to assess genotype-phenotype correlations. Mutant and wild type plasmids were constructed and transfected into HEK293T cells to evaluate LDLR mRNA and protein expression. In parallel, bioinformatics tools were employed to predict structural and functional changes in the mutant LDLR. Immunofluorescence analysis revealed no significant difference in the intracellular localization of the p.Gly343Ser mutation, whereas protein expression of the p.Ala627Thr mutation was decreased and predominantly localized in the cytoplasm. Western blotting has showed that protein expression levels of the mutant variants were markedly declined in both cell lysates and supernatants. Enzyme linked immunosorbent assay has demonstrated that LDLR protein levels in the supernatant of cell culture medium was not significant different from those of the wild-type group. However, LDLR protein levels in the cell lysate of both the Gly343Ser and Ala627Thr variants groups were significantly lower than those in the wild-type group. Bioinformatic predictions further suggested that these mutations may affect post-translational modifications of the protein, providing additional insight into the mechanisms underlying the observed reduction in protein expression. In this study, we identified two heterozygous pathogenic variants in the LDLR gene, c.G1027A (p.Gly343Ser) and c.G1879A (p.Ala627Thr), in a family with familial hypercholesterolemia. We also conducted preliminary investigations into the mechanisms by which these mutations contribute to disease pathology.

Optimization of Screening Media to Improve Antimicrobial Biodiscovery from Soils in Undergraduate/Citizen Science Research-Engaged Initiatives.

Background/Objectives: Research-engaged academic institutions offer the opportunity to couple undergraduate education/citizen science projects with antimicrobial biodiscovery research. Several initiatives reflecting this ethos have been reported internationally (e.g., Small World, Tiny Earth, MicroMundo, Antibiotics Unearthed). These programs target soil habitats due to their high microbial diversity and promote initial screening with non-selective, nutrient media such as tryptic soy agar (TSA). However, evaluation of published outputs to date indicates that isolate recovery on TSA is consistently dominated by the genera Bacillus, Pseudomonas, and Paenibacillus. In this study, we evaluated the potential of soil extract agar to enhance soil isolate diversity and antibiosis induction outcomes in our undergraduate Antibiotics Unearthed research program. Methods: We comparatively screened 229 isolates from woodland and garden soil samples on both tryptic soy agar (TSA) and soil extract agar (SEA) for antimicrobial activity against a panel of clinically relevant microbial pathogens. Results: On one or both media, 15 isolates were found to produce zones of clearing against respective pathogens. 16S rRNA gene sequencing linked the isolates with three genera: Streptomyces (7), Paenibacillus (6), and Pseudomonas (2). Six of the Streptomyces isolates and one Pseudomonas demonstrated antimicrobial activity when screened on SEA, with no activity on TSA. Furthermore, incorporation of the known secondary metabolite inducer N acetyl-glucosamine (20 mM) into SEA media altered the pathogen inhibition profiles of 14 isolates and resulted in broad-spectrum activity of one Streptomyces isolate, not observed on SEA alone. In conclusion, SEA was found to expand the diversity of culturable isolates from soil and specifically enhanced the recovery of members of the genus Streptomyces. SEA was also found to be a superior media for antibiosis induction among Streptomyces isolates when compared to TSA. It was noted that Paenibacillus isolates' antibiosis induction demonstrated a strain-specific response with respect to the growth media used. Conclusions: The authors propose SEA inclusion of in soil screening protocols as a cost-effective, complementary strategy to greatly enhance outcomes in undergraduate/citizen science-engaged antimicrobial biodiscovery initiatives.

Insect frass as a promising remedy of increasing the fertility of agricultural land in Siberia

One of the main tasks of modern organic crop production is the search for effective environmentally friendly fertilizers. A promising approach to this problem could be the study insect frass, a product of organic processing by insects. Insect frass is obtained as secondary products from the cultivation of such economic insects as, for example, housefly (Musca domestica) and black soldier fly (Hermetia illucens). Insect frass has a high concentration of nitrogen, which was processed within the digestive tract of insects, which makes it more accessible to plants and contributes to their active distribution and development. It also has insecticidal and fungicidal properties due to its own microbiome. Insect frass can also enhance the moisture-holding capacity of the environment and be used as sorbents for petroleum products. The actual study examines the use of insect frass as a stand-alone growing medium or as one component of a finished substrate. A study was carried out and the nitrogen content in ammonium and nitrate forms, total acidity and the phytotoxicity index of insect frass in mixtures with lowland peat were measured. The study showed that insect frass has acceptable acidity for use as a substrate, as well as a high nitrogen content in two forms. At high concentrations, insect frass is a source of phytotoxic properties, therefore, when using it, it is necessary to maintain the ratio with other components of the substrate. Thus, insect frass, with normal application, can become an effective fertilizer for all household producers engaged in organic farming, where the tools for increasing productivity are seriously limited, and all useful elements taken from the soil must be replenished without the use of chemical fertilizers.

A liquid static culture using a gas-permeable film bag contributes to microbiology

We propose a simple tool for liquid static culture using a copolymer film with high gas permeability. The film bags were successfully used to culture microorganisms Escherichia coli, Komagataella phaffii (methylotrophic) and Bacillus sp. (biofilm-forming), with cells cultured under physical stress-free conditions with sufficient oxygen supply. Similar growth curves and plasmid productivity were observed for liquid shake and film bag E. coli cultures. The early growth response of the film bag culture following colony inoculation of liquid media differed from conventional shake cultures. Our results indicate that a gas-permeable film bag is a promising liquid culture tool and provides novel microbiology materials.

Unlocking sustainable solutions: wood waste as a novel substrate for polyhydroxybutyrate (PHB) production to combat plastic pollution

ABSTRACT Polyhydroxybutyrate (PHB) is considered as a hope for bioplastic production, which can serve as a sustainable alternative. Utilizing feedstock as substrate is widely explored for the production but wood waste, which is abundant in cellulose, hemicellulose and lignocellulose, has limited studies for PHB production. Herein, wood waste is used as a biobased feedstock Hydrolyses of wood waste was done using sulphuric acid (H2SO4) to break down of cellulose and hemicellulose into simple carbon forms. The hydrolysed product was analysed for sugar presence by quantitative and qualitative methods. Pseudomonas fluorescens bacterial strain was used for the production purpose using hydrolysed wood waste as substrate media. The Plackett-Burman design (PBD) and response surface methodology (RSM) were applied to optimize the growth media. The results of PBD were used to identify significant factors influencing PHB production, which were then further optimized using RSM. The work’s results conclusively demonstrated that P. fluorescens possesses the capability to effectively utilize wood waste and wastewater as substrate media up to production rate of 13–14 mg mL−1 of PHB. Fourier Transformed Infra-Red (FTIR) spectroscopic peaks confirm the produced product is PHB, which is a type of polyhydroxyalkanoate (PHA), classified within the polyester family highlighting wood waste potential as a sustainable solution to address plastic pollution.

Characterization of the dynamic viscosity of cell cultures and its effect on mixing performance in a spinner flask bioreactor

Computational fluid dynamics (CFD) models have been developed to simulate cell culturing bioreactors but assume water-like viscosity properties due to significant data gaps. This study characterized the dynamic viscosity of HEK-293 cell cultures and evaluated its effect on mixing performance in a spinner flask bioreactor. Viscosity measurements indicated that the cell culture media, media with microcarriers, and cell cultures presented shear thinning behaviors within the measured shear rate range of 1–100 s−1. The viscosity also increased with the microcarrier concentrations and growth of cell culture. The CFD model, incorporating dynamic viscosity data, showed that shear stress and Kolmogorov length profiles are significantly influenced by microcarrier concentrations and cell culture growth. Higher microcarrier concentrations led to higher average shear stress and Kolmogorov values. The cultured HEK-293 cells after seven days of growth also had higher average shear stress and Kolmogorov values than at the day of seeding, indicating an impact caused by the cells’ metabolism and biomass. Overall, the results indicated that assuming water-like properties underestimates shear stress and Kolmogorov length scales, especially at zones of lower shear rates due to the observed shear thinning behavior. Thus, careful monitoring of dynamic viscosity of cell cultures and proper control of mixing parameters are critical to deliver the desired mixing conditions for optimized cell growth especially during scale-up production operations.

Lubrication and antibacterial performance and mechanism of functionalized ionic liquids in glycerol media with different water contents

Glycerol lubricants face limitations in industrial applications due to their inadequate antimicrobial properties. In this study, a functionalized long-chain quaternary ammonium salt ionic liquid, BTA-16-BTA, was synthesized. The antimicrobial and tribological properties of BTA-16-BTA as a lubricant additive in glycerol media with different water contents were investigated. The results show that the antimicrobial performance of the glycerol medium is positively correlated with the amount of BTA-16-BTA added. At the optimal concentration of 1.5 % BTA-16-BTA dispersion, excellent friction-reducing, anti-wear, and extreme pressure properties were observed. The friction-reducing and anti-wear effects were further enhanced when 1 % water was added to the system. The liquid medium containing BTA-16-BTA can inhibit the growth of Escherichia coli, indicating that BTA-16-BTA possesses antimicrobial properties.

Laser-driven synthesis of functional materials for advanced energy applications: A short review

Since the initial exploration of synthesis possibilities at a liquid–solid interface through pulsed laser irradiation of iron in water, the pulsed laser ablation in liquid media (PLAL) process has garnered significant attention for constructing various nanostructures due to its simple, ultrafast, and clean methodology that avoids byproduct formation. Recent advances in functionalized heterostructures via the PLAL process suggest a broad range of applications for energy- conversion systems, including water splitting and energy harvesting. This review surveys recent advancements in PLAL for energy applications, with a specific focus on its applications in oxygen evolution reaction (OER) electrocatalysts and energy harvesting devices. The review discusses the synthesis mechanisms, advantages, challenges, and future research directions of PLAL, aiming to provide a comprehensive understanding of its potential and limitations.

Study on the feasibility of using livestock blood as a fetal bovine serum substitute for cultured meat.

Fetal bovine serum (FBS) accounts for the largest portion of the cost of cultured meat production or cell culture experiments and is highly controversial in terms of animal welfare because it is taken from the fetus of a pregnant cow during slaughtering. Nevertheless, FBS is the most important supplement in the cell culture manufacturing process. This study aimed to develop an FBS substitute from slaughterhouse waste blood to reduce the cost of FBS in cultured meat production through various experiments. Our study successfully demonstrated that adult livestock blood obtained from slaughterhouses can effectively replace FBS. Our substitute, when cultured with bovine myosatellite cells, demonstrated cell growth that was either equivalent to or superior to that of commercial FBS. In the process of muscle generation through differentiation, the substitutes from bovine and chicken formed 70%-75% more bovine muscle compared to the control group using FBS. Furthermore, using the FBS substitute can reduce cell culture costs by approximately 61% compared to using commercial FBS. Therefore, the groundbreaking FBS substitute will not only contribute to the development of technology to mass-produce cultured meat using livestock byproducts but will also lower the production cost of media for experimental cell culture or vaccine production.

Biological conversion of lignin-derived ferulic acid from wheat bran into vanillin

Lignin is a promising feedstock for producing vanillin, one of the most extensively used flavor enhancers. However, the biotransformation performance of lignin derivatives into vanillin is still unsatisfactory. In this study, an efficient conversion strategy of lignin into vanillin was established by employing engineered Saccharomyces cerevisiae as a whole-cell biocatalyst. Optimization of cell culture media and whole-cell bioconversion improved the production efficiency of vanillin. The vanillin titer reached 15.3 mM with a molar yield of 71 % in fed-batch fermentation mode, while incorporating in-situ product separation, demonstrated a remarkable 2.6-fold increase. The whole-cell bioconversion, coupled with in-situ separation, successfully converted real lignin hydrolysate into a record vanillin titer of 21.1 mM, equivalent to 1.8 mg of vanillin per gram of wheat bran biomass. The whole-cell bioconversion process integrated in-situ product separation, represents a sustainable approach for vanillin production and offers a promising pathway for lignin valorization.