Biotechnology and Bioprocess Engineering最新文献

Epilepsy, a predominant neurological disorder affecting about 1% of the worldwide population, demands effective treatment options. An antiepileptic drug called brivaracetam has proven amazing efficacy in preventing epilepsy progression, garnering attention for novel synthesis methods. Despite recent progress in conventional synthesis routes, challenges such as expensive catalysts, inconvenient substrates, and hazardous solvents persist. In this context, we share the first finding that immobilized penicillin G acylase (IPGA) can catalyze the polarity reversal conjugate addition reaction. This synthesis is straightforward and does not require any purification. Yield up to 92.41% was achieved at 55 °C using dimethyl sulfoxide as a solvent. The catalytic specificity of IPGA was demonstrated through control experiments. Nonetheless, this research demonstrates the potential of IPGA and other biocatalysts to enable sustainable and effective organic synthesis processes and showcase the promiscuity of existing enzymes.

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Gossypium hirsutum L., commonly known as upland cotton, is cultivated globally for natural fiber, feed, and seed oil. To enhance agricultural productivity and quality, living modified (LM) cotton has been developed and utilized since the late 1990s. Due to environmental concerns, such as biodiversity risks associated with living modified organisms (LMOs), the Korean government, including the Ministry of Environment, has been conducting LMO natural environment monitoring and post-management projects. In this study, we developed a loop-mediated isothermal amplification (LAMP) assay to detect five specific LM cotton events (MON88701, MON531, MON15985, MON88913, and COT102), which were the most abundant volunteers observed from 2019 to 2021 in South Korea. The event-specific LAMP assays for the five LM cotton events were established with a 40-min reaction time using LAMP reaction buffer, Bst DNA polymerase, and event-specific primers. The limit of detection ranged from 0.01 to 1 ng/μL. The assays' specificity and sensitivity were validated through colorimetric changes, fluorescence intensity measurements, and conventional PCR. To demonstrate the practical application of the LAMP assays, we tested 22 LM cotton volunteers collected from the natural environment in 2021. Each LAMP assay event-specifically amplified the respective LM cotton volunteers. These results indicate that the developed LAMP assays are effective tools for the efficient management and detection of LM cotton in field surveys, supporting regulatory compliance and environmental monitoring efforts.

As azo dyes are commercially used to treat textiles and leather articles, but they were toxic, allergenic, carcinogenic, and mutagenic to human and environment if they were not well treated, the content and degradation of azo dyes in wastewater were very important. So far, various methods such as physical, chemical, and biological methods have been applied to solve this. As a sequential work, we tried to remove a famous azo dye, Reactive Black 5 (RB5) by biological assay and found that Clostridium acetobutylicum showed the best decolorization of RB5 among the hydrogen producing Clostridium species examined. It was also found 100 ppm concentration of RB5 did not affect biohydrogen production of C. acetobutylicum suggesting simultaneous degradation of azo dye and biohydrogen production was possible. Optimal condition for simultaneous decolorization and biohydrogen production was selected as pH 6, 40 °C, and 200 rpm, and high concentration (200 ppm) of RB5 could be decolorized up to 97%. When C. acetobutylicum was examined for further decoloring activities, it was showed the decolorization of various dyes such as Bromophenol, Bromocresol, Methyl Red, and Reactive Red 120. This study showed potential application of C. acetobutylicum in wastewater treatment by simultaneous decolorizing and biohydrogen production.

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The serum carcinoembryonic antigen (CEA) level lacks the specificity required for precise colorectal cancer (CRC) diagnosis. To address this limitation, we investigated CEA-expressing circulating epithelial cells (CCECs) as a diagnostic biomarker for CRC. Human peripheral blood mononuclear cells, obtained from patient samples, were suspended in Matrigel and subjected to cyto-centrifugation for immunohistochemistry staining, enhancing cell immobilization on glass slides. Using this method, we successfully identified CCECs in 63% of CRC patients, whereas they were present in only 5% of non-cancer cohorts. Subsequently, receiver operating characteristic (ROC) analysis demonstrated the superior diagnostic capability of the CCEC count (area under the curve [AUC]-ROC: 0.817; p = 0.003) compared to serum CEA levels (AUC-ROC: 0.747; p = 0.021). Furthermore, a strong correlation emerged between tissue CEA expression and CCEC counts, with CCECs exclusively detectable in patients with CEA-positive tumors. The CCEC count also exhibited a high predictive value for estimating lymphovascular invasion and distant metastasis. To further enhance the clinical utility of CCECs, we established an integrated CEA score by combining serum CEA levels and CCEC counts. This scoring system exhibited superior AUC-ROCs for distinguishing malignancy and estimating tumor metastatic potential compared to individual biomarkers. Our study underscores the promise of CCECs as a biomarker for distinguishing and diagnosing CRC.

Quercetin (Q), a plant-derived flavonoid, has antibacterial properties against a wide range of bacterial types, and preferentially targets gram-positive bacteria. However, because of its lower activity than that of current antibiotics, Q has only been used as an antimicrobial adjuvant, particularly against antimicrobial-resistant strains, where it works synergistically with other agents to increase their antimicrobial activity and selectivity. Zinc oxide (ZnO) nanomaterials, which are among the most extensively researched and powerful antibacterial compounds, have limitations in terms of off-target effects and cytotoxicity at high concentrations and this necessitates the use of biocompatible chemicals for their modification. In this study, we bio-conjugated ZnO with Q using cell-free supernatant of Staphylococcus ATCC 25923. UV–visible spectroscopy, X-ray diffraction analysis, Fourier-transform infrared spectroscopy, dynamic light scattering, and transmission electron microscopy confirmed the formation of ZnO nanoparticles (NPs) with Q nanocomposite (ZnO@Q NC). The in vitro antibacterial activity of ZnO@Q NC against gram-negative and gram-positive bacterial species was evaluated as the minimum inhibitory concentration. The results demonstrated that ZnO@Q NC exhibited a 4–32-fold higher preferential activity toward Staphylococcus species than Q and ZnO NPs. Additional mechanistic studies revealed that ZnO@Q NC disrupted bacterial membranes and prevented biofilm formation. The biocompatibility of ZnO@Q NC with WI-38 cells was assessed, and the synergistic use of ZnO@Q NC with antibiotics was suggested to reduce its cytotoxicity. Overall, the results demonstrated that conjugating ZnO NPs with Q can significantly boost their bactericidal efficacy and selective pressure against Staphylococcus species.

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Cyanin chloride, one of the active ingredients in figs, is a glycoside made of two sugars linked to a cyanidin aglycone. Although many studies have reported on the skin efficacy of cyanidin aglycone, there have been few reports on human psoriasis to date. Therefore, we focused on excessive inflammation and loss of skin barrier function, which are the main characteristics of psoriasis, and verified the function of cyanin chloride on the psoriasis. Cyanine chloride removed DPPH and ABTS radicals in a concentration-dependent manner and significantly inhibited NO production in LPS-induced RAW264.7 cells as well as suppressed inflammatory cytokines such as iNOS, COX-2, IL-6, and IL-1α/β. Moreover, we used TNF-α/IL-17A/IFN-γ-induced HaCaT cells, a human skin cell model of psoriasis. Cyanin chloride significantly inhibited the mRNA expression of inflammatory cytokines, such as IL-1α, IL-1β, and IL-6, and chemokines CXCL8 and CCL20 in TNF-α/IL-17A/IFN-γ-induced HaCaT cells. Cyanin chloride significantly inhibited the phosphorylation of STAT3 transcription factor in a concentration-dependent manner, confirming the regulatory function of CCL20 chemokine. Finally, cyanin chloride significantly restored the TEER value in TNF-α/IL-17A/IFN-γ-induced HaCaT cells, confirming the effect of strengthening the skin barrier function. In addition, cyanin chloride increased the mRNA levels of filaggrin which is cornified envelope proteins of the epidermal layer, in a concentration-dependent manner in normal epidermal cells. Taken together, the above results suggest that cyanin chloride can be considered a natural candidate for improving psoriasis, an incurable skin disease, not only by lowering excessive skin inflammatory reactions but also by restoring the skin barrier.

Sugarcane molasses is one of the by-products from the sugar industry, which is a candidate carbon source for microbes to produce high-value docosahexaenoic acid (DHA) that is beneficial to human health. The aim of this study was to optimize the sugarcane molasses medium for DHA production by Aurantiochytrium sp. and investigate the treatment and disposal of sugarcane molasses fermentation wastewater. Firstly, the sugarcane molasses volume ratio, carbon–nitrogen ratio and citric acid concentration were optimized for the sugarcane molasses medium, and the DHA production in Aurantiochytrium sp. R2A35 was increased from 2.63 to 7.48 g/L. Then, the two-staged oxygen supply strategy was adopted in the expanded culture, and the DHA production reached to 10.04 g/L. And the DHA proportion in total lipid was significantly improved from 41.08 to 60.46% (w/w), which promoted the total DHA production. The data showed that high oxygen supply promoted biomass accumulation and low oxygen supply promoted DHA accumulation in Aurantiochytrium sp. R2A35. It was also found that Aurantiochytrium sp. R2A35 preferred to use the reducing sugar in sugarcane molasses and then used sucrose. The final utilization rate of reducing sugar was 100% and the utilization rate of sucrose was 94.37%. Finally, the concentrated fermentation wastewater, sugarcane leaves and bagasse were mixed as a ratio of 2:5:12 for composting, and the compost product could meet the commercial organic fertilizer standard. This study accelerated the resource utilization of sugarcane waste and provided an alternative solution for DHA industry.

Cancer is one of the most common health problems affecting individuals worldwide. In the field of biomedical engineering, one of the main methods for cancer diagnosis is the analysis of histological images of tissue structures and cell nuclei using artificial intelligence. Here, we compared the performance of 15 deep learning methods viz: UNet, Deep-UNet, UNet-CBAM, RA-UNet, SA-Unet and Nuclei-SegNet, UNet-VGG2016, UNet-Resnet-101, TransResUNet, Inception-UNet, Att-UNet++ , FF-UNet, Att-UNet, Res-UNet and a new model, DanNucNet, in pathological nuclei segmentation on tissue slices from different organs on five open datasets: MoNuSeg, CoNSeP, CryoNuSeg, Data Science Bowl, and NuInsSeg. Before training on the data, the pixel intensity and color distribution were analyzed, and different augmentation techniques were applied. The results showed that the UNet-based model with 34.57 million Deep-UNet parameters performed the best, outperforming all models in terms of the Dice coefficient from 3.13 to 22.91%. The implementation of Deep-UNet in this context provides a valuable tool for accurate extraction of cancer cell nuclei from histological images, which in turn will contribute to further developments in cancer pathology and digital histology.

The chemical, pharmaceutical, and cosmetic industries are currently confronted with the challenge of transitioning from traditional chemical processes to more sustainable biocatalytic methods. To support that aim, we developed various heterogeneous biocatalysts for an industrially relevant enzyme called oleate hydratase that converts oleic acid to 10-hydroxystearic acid, a fatty emollient substance useful for various technical applications. We used cheap support matrices such as silica, chitosan, cellulose, and agarose for further scale-up and economic feasibility at the industrial level alongside more sophisticated supports like metal–organic frameworks. Different physical and chemical binding approaches were employed. Particularly, by immobilizing oleate hydrates on a 3-aminopropyltriethoxysilane surface-functionalized cellulose matrix, we developed an enzyme immobilizate with almost 80% activity of the free enzyme. The long-term goal of this work was to be able to use the developed heterogeneous biocatalyst for multiple reuse cycles enabling profitable biocatalysis. Despite high initial conversion rate by the developed cellulose-based immobilizate, a depletion in enzyme activity of immobilized oleate hydratase was observed over time. Therefore, further enzyme modification is required to impart stability, the optimization of operational conditions, and the development of carrier materials that enable economical and sustainable enzymatic conversion of oleic acid to meet the commercial demand.

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