Increase CO2 recycling of Escherichia coli containing CBB genes by enhancing solubility of multiple expressed proteins from an operon through temperature reduction.

IMPORTANCE: In a previous study, we successfully engineered Escherichia coli capable of endogenous CO2 recycling through the heterologous expression of the Calvin-Benson Bassham genes. Establishing an efficient gene expression environment for recombinant strains is crucial, on par with the importance of metabolic engineering design. Therefore, the primary objective of this study was to further mitigate greenhouse gas emissions by investigating the effects of culture temperature on the formation of inclusion bodies (IB) and CO2 fixation activity in the engineered bacterial strain. The findings demonstrate that lowering the culture temperature effectively suppresses IB formation, enhances CO2 recycling, and concurrently increases the accumulation of organic acids. This temperature control approach, without adding or modifying compounds, is both convenient and efficient for enhancing CO2 recycling. As such, additional optimization of various environmental parameters holds promise for further enhancing the performance of recombinant strains efficiently.

Enhancing biodegradation of PBAT through bio-stimulation using Pseudozyma jejuensis for effective plastic waste reduction.

Polybutylene adipate-co-terephthalate (PBAT) is a flexible and biodegradable material that finds applications in mulching film and the food packaging industry. In this study, we aimed to address the global plastic waste problem by developing an improved biodegradation system for PBAT. Our focus was on utilizing the biodegradation capabilities of Pseudozyma jejuensis, a microorganism known for its ability to decompose Polycaprolactam (PCL). Through bio-stimulation, we aimed to enhance the growth mechanism of P. jejuensis and optimize PBAT biodegradation. Our results demonstrated significant structural changes in the PBAT film, as revealed by FT-IR analysis. Moreover, FE-SEM imaging exhibited evident surface erosion and pitting, indicating physical alterations due to biodegradation. These findings provide strong evidence for the efficiency of our developed biodegradation system. To fully harness the potential of this system and enable its practical implementation, further research is warranted to optimize and scale up the process. Our work contributes to the ongoing efforts to combat the global plastic waste crisis, offering a valuable solution for the efficient biodegradation of PBAT.

Detection of triclosan using 2-domain hemoglobin promoter of Daphnia magna.

Triclosan has been widely used as an antimicrobial agent. However, triclosan was found to cause toxicity, including muscle contraction disturbances, carcinogenesis, and endocrine disorders. In addition, it was found to affect central nervous system function adversely and even have ototoxic effects. Conventional methods for detecting such triclosan can be performed easily. However, the conventional detection methods are inadequate in precisely reflecting the impact of toxic substances on stressed organisms. Therefore, a test model for the toxic environment at the molecular level through the organism is needed. From that point of view, Daphnia magna is being used as a ubiquitous model. D. magna has the advantages of easy cultivation, a short lifespan and high reproductive capacity, and high sensitivity to chemicals. Therefore, the protein expression pattern of D. magna that appear in response to chemicals can be utilized as biomarkers for detecting specific chemicals. In this study, we characterized the proteomic response of D. magna following triclosan exposure via two-dimensional (2D) gel electrophoresis. As a result, we confirmed that triclosan exposure completely suppressed D. magna 2-domain hemoglobin protein and evaluated this protein as a biomarker for triclosan detection. We constructed the HeLa cells in which the GFP gene was controlled by D. magna 2-domain hemoglobin promoter, which under normal conditions, expressed GFP, but upon triclosan exposure, suppressed GFP expression. Consequently, we consider that the HeLa cells containing the pBABE-HBF3-GFP plasmid developed in this study can be used as novel biomarkers for triclosan detection.

Detection of Salmonella dublin using the vitellogenin 2 promoter of Daphnia magna.

Salmonella is a well-known bacterium that causes waterborne diseases in humans and primates. The need for test models to detect such pathogens and study the responses of such organisms to induced toxic environments is vital. Daphnia magna has been ubiquitously used in aquatic life monitoring for decades because of outstanding properties, such as facile cultivation, short lifespan, and high reproductive capacity. In this study, the proteomic response of D. magna exposed to four Salmonella strains (Salmonella dublin, Salmonella enteritidis, Salmonella enterica, and Salmonella typhimurium) was characterized. As indicated by two-dimensional gel electrophoresis, vitellogenin fused with superoxide dismutase was completely suppressed under exposure to S. dublin. Thus, we evaluated the feasibility of using the vitellogenin 2 gene as a biomarker for S. dublin detection, particularly in providing rapid, visual detection through fluorescent signals. Accordingly, the applicability of the HeLa cells transfected with pBABE-Vtg2B-H2B-GFP as a biomarker for the detection of S. dublin was evaluated, and it was confirmed that the fluorescence signal decreased only when S. dublin was treated. Therefore, such HeLa cells can be utilized as a novel biomarker for detecting S. dublin.

Optimized Rhodobacter sphaeroides for the Production of Antioxidants and the Pigments with Antioxidant Activity.

The photosynthetic bacterium, Rhodobacter sphaeroides, is a bacterium that can grow in a variety of environments and produces substances with antioxidant effects. In this study, we investigated the culture conditions to increase the production of antioxidants in R. sphaeroides, which can grow under both aerobic and anaerobic conditions. After incubation in the exponential phase and stationary phase under both conditions, a 2,2-diphenyl-1-picrylhydrazyl assay was used to confirm the antioxidant effect. Although the highest antioxidant effect was shown in the stationary phase under aerobic conditions, the antioxidant effect of each cell was found to be greater when cultured under anaerobic conditions. The antioxidant activity of R. sphaeroides was increased by sonication. In addition, the contents of carotenoids and bacteriochlorophyll, which are pigments with antioxidant effects, produced by R. sphaeroides were measured. We confirmed that the content of carotenoids was higher in anaerobic conditions than in aerobic conditions. However, when measuring the content of the bacterium, we found that there was more content in aerobic conditions. Therefore, we confirm that when grown in anaerobic conditions, the antioxidant effect of R. sphaeroides is higher, which suggests that this antioxidant effect comes from the effect of carotenoid.

Evaluation of Growth and Utilization Potential of Rhodobacter sphaeroides in Reused Medium.

Rhodobacter sphaeroides is a metabolically versatile purple non-sulfur bacteria that can produce valuable substances. As the low-cost and high-efficiency production of valuable substances is attracting attention, the reuse of the medium is emerging as a promising strategy. Therefore, in this study, the growth of R. sphaeroides was evaluated by reusing the medium of Escherichia coli and Saccharomyces cerevisiae. As a result, in the reuse of the medium in which S. cerevisiae was cultured, sufficient growth of R. sphaeroides could be confirmed, and especially, the growth of R. sphaeroides was not inhibited under aerobic conditions. Therefore, it is considered that the strategy of reusing the medium of S. cerevisiae is sufficiently feasible. Of the organic compounds investigated, R. sphaeroides grew best in succinic acid, followed by malic acid, citric acid, acetic acid, and glucose. In addition, by comparing photopigment synthesis in the reused medium, we propose the hypothesis that succinic acid may play an important role in photopigment synthesis for the first time.

Comparison of polylactic acid biodegradation ability of Brevibacillus brevis and Bacillus amyloliquefaciens and promotion of PLA biodegradation by soytone.

Polylactic acid (PLA), a biodegradable plastic, is used to substitute commercial plastics in various fields such as disposable packaging materials and mulching films. Although the biodegradation of PLA under submerged or composting conditions is accelerated, increasing the biodegradability of PLA under soil burial conditions is still a challenge. This study reviews and compares the PLA biodegradation ability of Bacillus amyloliquefaciens and Brevibacillus brevis, both PLA-degrading bacteria. The biodegradation ability of a single bacteria in non-composting conditions was evaluated. In addition, in terms of biostimulation, PLA biodegradation according to nitrogen sources was compared. As a result, a higher PLA biodegradation ability was found in B. brevis than in B. amyloliquefaciens. Moreover, it was confirmed that the biodegradation of the PLA film was increased by using soytone as a nitrogen source in both bacteria. Controlling the nitrogen source could be a new way to increase the biodegradation of PLA.

Isoprene production by Rhodobacter sphaeroides and its antimicrobial activity.

Extensive studies on the antimicrobial activity of terpene-based substances, which are the main components of essential oils, are continuously underway. And some hydrocarbons constituting antimicrobial substances have been reported to exhibit the antimicrobial activity. This study confirmed the production of isoprene, the most basic constituent hydrocarbon of terpene, by Rhodobacter sphaeroides, and investigated the antimicrobial activity of isoprene and its mechanism. We developed an air-sharing culture system in which different bacterial cultures aseptically shared the same atmosphere, to evaluate the effect of volatile isoprene. Effects were tested on two Gram-negative bacteria, and on two Gram-positive bacteria. As a result, the isoprene released from R. sphaeroides showed the antimicrobial activity against all evaluated strains, especially against Gram-positive bacteria than Gram-negative bacteria. In addition, the microstructure of the bacteria was evaluated via FE-SEM. The FE-SEM images showed that isoprene has the antimicrobial activity mechanism that causes cell death by acting on the cell wall or the extracellular membrane.

Evaluation of muscle damage using ultrasound imaging.

[Purpose] This study aimed to quantitatively analyze characteristics of and changes in internal muscle structure according to the time of delayed onset muscle soreness (DOMS) using ultrasound imaging, thereby presenting clinical evidential data for evaluation of muscle damage. [Subjects] We recruited 38 male subjects. [Methods] Ultrasound images of the medial gastrocnemius muscle prior to induction of DOMS and immediately after, 24 hours after, 48 hours after, and 72 hours after induction of DOMS were obtained, and the thickness and pennation angle of the muscle were measured. [Results] The muscle thickness gradually increased until 48 hours after induction of DOMS and decreased after 72 hours. The pennation angle also gradually increased until 48 hours after induction of DOMS and decreased after 72 hours. [Conclusion] Ultrasound imaging is considered useful for assessment of structural characteristics of muscles when muscle damage like DOMS takes place.

Estimation of mercury speciation in soil standard reference materials with different extraction methods by ion chromatography coupled with ICP-MS.

Analytical methods for the speciation of mercury, based on microwave extraction and sonication extraction, have been tested to determine the inorganic mercury and methyl mercury contents in two standard soil reference materials: SRM 2710 Montana Soil and BCR 580 estuarine sediment. Prior to applying the speciation extraction methods, the mineral compositions were analyzed via XRD analysis, with SRM 2710 shown to be composed mostly of aluminum silicate minerals, while carbonate minerals were the major constituent in BCR 580. Two extraction methods, microwave and sonication, were tested for the analysis and recovery efficiency of total mercury. The accuracy and efficiency of each extraction method was also compared. In the analysis of total mercury, the microwave extraction method, with using methanol and HCl as extractants, was better for SRM2710, while the application of the sonication extraction method was more efficient for the calcite-based BCR 580. The results showed good separation and recovery efficiencies, with values reaching 100% of those estimated. The sonication method was selected for the speciation of mercury, especially in BCR 580. An extraction solution comprising of a 1:1 mixture of methanol and HCl was used for the sonication extraction of BCR 580, with the resulting extractants analyzed by IC-HG-ICP-MS for methyl mercury and inorganic mercury. As a simple, rapid, sensitive, and accurate method, sonication extraction was found to be satisfactory.

Erectile potentials of a new phosphodiesterase type 5 inhibitor, DA-8159, in diet-induced obese rats.

AIM: To examine the changes in the erectile function in diet-induced obese rats and investigate the oral efficacy of DA-8159, a new phosphodiesterase type 5 (PDE5) inhibitor, on penile erection in obese rats. METHODS: The rats were fed a high-energy diet for 12 weeks and divided into three groups: an obesity-resistant (OR) control group, an obesity-prone (OP) control group, and an OP-DA-8159 treatment (DA-8159) group. The electrostimulation-induced erectile responses were measured in all groups. The body weight, plasma cholesterol, triglyceride and glucose levels were also measured. RESULTS: In the OP control group, the maximum intracavernous pressure (ICP) and ICP/blood pressure (ICP/BP) ratio after electric stimulation were significantly lower than those in OR control group. The corresponding area under the curve (AUC) of the ICP/BP ratio, the detumescence time and the baseline cavernous pressure were also lower than those in the OR control group, but this difference was not significant. The body weight gain, plasma cholesterol and triglyceride level in the OP group were significantly higher than those in the OR group. After administering the DA-8159, a significant increase in the maximum ICP and the ICP/BP ratio were observed. The corresponding AUCs in the DA-8159 group were also higher than those in the two control groups. Furthermore, the detumescence time was significantly prolonged after treatment with DA-8159. CONCLUSION: These results demonstrate that diet-induced obesity affects the erectile function in rats and these erectile dysfunction (ED) can be improved by the treatment with DA-8159, indicating DA-8159 might be a treatment option for ED associated with obesity.