Biotechnology Journal最新文献

Industrial production of bacterial cellulose (BC) remains challenging due to significant production costs, including the choice of appropriate growth media. This research focuses on optimization of cheese whey (CW) based media for enhanced production of BC. Two modifications were made for CW medium for BC production with Komagataeibacter rhaeticus MSCL 1463. BC production in a medium of enzymatically hydrolyzed CW (final concentration of monosaccharides: glucose 0.13 g L⁻¹, galactose 1.24 g L⁻¹) was significantly enhanced, achieving a yield of 4.95 ± 0.25 g L⁻¹, which markedly surpasses the yields obtained with the standard Hestrin-Schramm (HS) medium containing 20 g L⁻¹ glucose and acid-hydrolyzed CW (final concentration of monosaccharides: glucose 1.15 g L⁻¹, galactose 2.01 g L⁻¹), which yielded 3.29 ± 0.12 g L⁻¹ and 1.01 ± 0.14 g L⁻¹, respectively. We explored the synergistic effects of combining CW with various agricultural by-products (corn steep liquor (CSL), apple juice, and sugar beet molasses). Notably, the supplementation with 15% corn steep liquor significantly enhanced BC productivity, achieving 6.97 ± 0.17 g L⁻¹. A comprehensive analysis of the BC's physical and mechanical properties indicated significant alterations in fiber diameter (62–167 nm), crystallinity index (71.1–85.9%), and specific strength (35–82 MPa × cm³ g⁻¹), as well as changes in the density (1.1–1.4 g cm⁻³). Hydrolyzed CW medium supplemented by CSL could be used for effective production of BC.

The ELISA is the most worldwide method for immunoassay. However, the ELISA is losing ground due to low reproducibility of manual experimental processes in both R&D and IVD areas. An automated platform is a good solution, but there are still limitations owning to extremely high cost and requiring large space to set up especially for a small size laboratory. Here, we present a novel all-in-one platform called “VEUS” settable on the laboratory table that offers comprehensive automation of the entire multiplex immunoassay process by exploiting antibody conjugated magnetic particles, quality control and then immunoanalytical reaction, thereby enhancing detection sensitivity and high reproducibility. As a proof of concept, the system exhibits a sensitive LOD of 0.6 and 3.1 pg mL⁻¹ within 1 h run, comparable precision that of molecular diagnostic systems based on PCR method, enabling rapid multiplex diagnosis of Influenza A, Influenza B, and COVID-19 viruses with similar symptoms. Through automation by the all-in-one system, it can be used by novice users, something innovative for immunoassays, relying heavily on user experience. Furthermore, it can contribute to streamline entire immunoassay processes of diverse biomarkers with high reproducibility and convenience in laboratories.

Daptomycin, a lipopeptide comprising an N-decanoyl fatty acyl chain and a peptide core, is used clinically as an antimicrobial agent. The start condensation domain (dptC1) is an enzyme that catalyzes the lipoinitiation step of the daptomycin synthesis. In this study, we integrated enzymology, protein engineering, and computer simulation to study the substrate selectivity of the start condensation domain (dptC1) and to screen mutants with improved activity for decanoyl loading. Through molecular docking and computer simulation, the fatty acyl substrate channel and the protein–protein interaction interface of dptC1 are analyzed. Key residues at the protein–protein interface between dptC1 and the acyl carrier were mutated, and a single-point mutant showed more than three-folds improved catalytic efficiency of the target n-decanoyl substrate in comparing with the wild type. Moreover, molecular dynamics simulations suggested that mutants with increased catalytic activity may correlated with a more “open” and contracted substrate binding channel. Our work provides a new perspective for the elucidation of lipopeptide natural products biosynthesis, and also provides new resources to enrich its diversity and optimize the production of important components.

Artificial Intelligence (AI) technology is spearheading a new industrial revolution, which provides ample opportunities for the transformational development of traditional fermentation processes. During plasmid fermentation, traditional subjective process control leads to highly unstable plasmid yields. In this study, a multi-parameter correlation analysis was first performed to discover a dynamic metabolic balance among the oxygen uptake rate, temperature, and plasmid yield, whilst revealing the heating rate and timing as the most important optimization factor for balanced cell growth and plasmid production. Then, based on the acquired on-line parameters as well as outputs of kinetic models constructed for describing process dynamics of biomass concentration, plasmid yield, and substrate concentration, a machine learning (ML) model with Random Forest (RF) as the best machine learning algorithm was established to predict the optimal heating strategy. Finally, the highest plasmid yield and specific productivity of 1167.74 mg L⁻¹ and 8.87 mg L⁻¹/OD₆₀₀ were achieved with the optimal heating strategy predicted by the RF model in the 50 L bioreactor, respectively, which was 71% and 21% higher than those obtained in the control cultures where a traditional one-step temperature upshift strategy was applied. In addition, this study transformed empirical fermentation process optimization into a more efficient and rational self-optimization method. The methodology employed in this study is equally applicable to predict the regulation of process dynamics for other products, thereby facilitating the potential for furthering the intelligent automation of fermentation processes.

This article primarily introduces a new treatment for liver fibrosis/cirrhosis. We developed a hepatic patch by combining decellularized liver matrix (DLM) with the hepatocyte growth factor (HGF)/heparin–complex and evaluated its restorative efficacy. In vitro prophylactic results, the HGF/heparin–DLM patches effectively mitigated CCl₄-induced hepatocyte toxicity and restored the cytotoxicity levels to the baseline levels by day 5. Furthermore, these patches restored albumin synthesis of injured hepatocytes to more than 70% of the normal levels within 5 days. In vitro therapeutic results, the urea synthesis of the injured hepatocytes reached 91% of the normal levels after 10 days of culture, indicating successful restoration of hepatic function by the HGF/heparin–DLM patches in both prophylactic and therapeutic models. In vivo results, HGF/heparin–DLM patches attached to the liver and gut exhibited a significant decrease in collagen content (4.44 times and 2.77 times, respectively) and an increase in glycogen content (1.19 times and 1.12 times, respectively) compared to the fibrosis group after 1 week, separately. In summary, liver function was restored and inflammation was inhibited through the combined effects of DLM and the HGF/heparin–complex in fibrotic liver. The newly designed hepatic patch holds promise for both in vitro and in vivo regeneration therapy and preventive health care for liver tissue engineering.

All-trans retinoic acid (atRA) is an endogenous ligand of the retinoic acid receptors, which heterodimerize with retinoid X receptors. AtRA is generated in tissues from vitamin A (retinol) metabolism to form a paracrine signal and is locally degraded by cytochrome P450 family 26 (CYP26) enzymes. The CYP26 family consists of three subtypes: A1, B1, and C1, which are differentially expressed during development. This study aims to develop and validate a high throughput screening assay to identify CYP26A1 inhibitors in a cell-free system using a luminescent P450-Glo assay technology. The assay performed well with a signal to background ratio of 25.7, a coefficient of variation of 8.9%, and a Z-factor of 0.7. To validate the assay, we tested a subset of 39 compounds that included known CYP26 inhibitors and retinoids, as well as positive and negative control compounds selected from the literature and/or the ToxCast/Tox21 portfolio. Known CYP26A1 inhibitors were confirmed, and predicted CYP26A1 inhibitors, such as chlorothalonil, prochloraz, and SSR126768, were identified, demonstrating the reliability and robustness of the assay. Given the general importance of atRA as a morphogenetic signal and the localized expression of Cyp26a1 in embryonic tissues, a validated CYP26A1 assay has important implications for evaluating the potential developmental toxicity of chemicals.

Tang, T., Xu, Y., Wang, L., & Zhang, P. (2023). In vitro acne disease model from inertial focusing effect for studying the interactions between sebocyte glands and macrophages. Biotechnology Journal, 18, e2300108. https://doi.org/10.1002/biot.202300108

In “ACKNOWLEDGMENTS” section, the text “The SZ95 cells and CBD in work is kindly provided by Yunnan Baiyao Group Co. Ltd.” was incorrect. We purchased the SZ95 cells from Yuchi Cell Biological Technology Co. Ltd. This should have read: “The CBD in work is kindly provided by Yunnan Baiyao Group Co. Ltd.” In “2.1 cell culture” section, the text “SZ95 cells were purchased from Yuchi Cell Biological Technology Co. Ltd.” should be added on the beginning of paragraph.

We apologize for this error.

Alzheimer's disease (AD), the most common form of dementia, has gotten considerable attention. Previous studies have demonstrated that clioquinol (CQ) as a metal chelator is a potential drug for the treatment of AD. However, the mode of action of CQ in AD is still unclear. In our study, the antioxidant effects of CQ on yeast cells expressing Aβ42 were investigated. We found that CQ could reduce Aβ42 toxicity by alleviating reactive oxygen species (ROS) generation and lipid peroxidation level in yeast cells. These alterations were mainly attributable to the increased reduced glutathione (GSH) content and independent of activities of superoxide dismutase (SOD) and/or catalase (CAT). CQ could affect antioxidant enzyme activity by altering the transcription level of related genes. Interestingly, it was noted for the first time that CQ could combine with antioxidant enzymes to reduce their enzymatic activities by molecular docking and circular dichroism spectroscopy. In addition, CQ restored Aβ42-mediated disruption of GSH homeostasis via regulating YAP1 expression to protect cells against oxidative stress. Our findings not only improve the current understanding of the mechanism of CQ as a potential drug for AD treatment but also provide ideas for subsequent drug research and development.

IMP (inosinic acid) is a crucial intermediate in the purine metabolic pathway and is continuously synthesized in all cells. Besides its role as a precursor for DNA and RNA, IMP also plays a critical or essential role in cell growth, energy storage, conversion, and metabolism. In our study, we utilized the circularly permuted fluorescent protein (cpFP) and IMP dehydrogenase to screen and develop the IMP biosensor, IMPCP1. By introducing a mutation in the catalytically active site of IMPCP1, from Cys to Ala, we disrupted its ability to catalyze IMP while retaining its capability to bind to IMP without affecting the IMP concentration in the sample. To immobilize IMPCP1, we employed the SpyCatcher/SpyTag system and securely attached it to Magarose-Epoxy, resulting in the development of the IMP rapid test kit, referred to as IMPTK. The biosensor integrated into IMPTK offers enhanced stability, resistance to degradation activity, and specific recognition of IMP. It is also resistant to peroxides and temperature changes. IMPTK serves as a rapid and stable assay for analyzing IMP concentrations in fermentation broth. Within the linear range of IMP concentrations, it can be utilized as a substitute for HPLC. The IMPTK biosensor provides a reliable and efficient alternative for monitoring IMP levels, offering advantages such as speed, stability, and resistance to environmental factors.

Tang, T., Wei, Y., Jia, H., Wang, L., Xu, Y. e., & Zhang, P. (2023). 3D artificial sebocyte glands from inertial focusing effect for facile and flexible analysis of light damage and drug screening. Biotechnology Journal, 18, e2200634. https://doi.org/10.1002/biot.202200634

In “2.1 cell culture” section, the text “Human immortalized SZ95 sebocytes (granted from Yunnan Baiyao Group Co., Ltd.) were cultured in a 35 mm petri dish (Corning, New York, USA) at 37°C in a humidified atmosphere containing 5% CO₂.” was incorrect. We purchased the SZ95 cells from Yuchi Cell Biological Technology Co. Ltd. This should have read: “Human immortalized SZ95 sebocytes (purchased from Yuchi Cell Biological Technology Co. Ltd) were cultured in a 35 mm petri dish (Corning, New York, USA) at 37°C in a humidified atmosphere containing 5% CO₂.”

We apologize for this error.