BIOCHEMICAL ENGINEERING JOURNAL
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2024
·
IF 3.7
Shu-Ping Zou,Wei Ding,Yu-Yue Han,Kun Niu,Ya-Ping Xue,Yu-Guo Zheng
L-Phosphinothricin (L-PPT) is the active compound of a broad-spectrum herbicide. In this study, we biosynthesized L-PPT via the asymmetric reduction of 2-carboxy-4-(hydroxymethylphosphoryl) butyric acid (PPO) using a whole-cell biocatalyst. Glutamate dehydrogenase from Pseudomonas moorei ( Pm GluDH), Glucose dehydrogenase from Exiguobacterium sibiricum ( Es GDH) and NAD kinase from Corynebacterium glutamicum ( Cg NADK) were coexpressed in E. coli , significantly increasing the catalytic efficiency by regenerating more NADPH to promote PPO reduction. Following the optimization of balancing the co-expression of the three enzymes in pETDuet-1 and pCDFDuet-1, the coexpressing strain increases in the intracellular NADP(H) concentration to 54.9 μmol/g DCW, which was 3.8-fold higher than the control. The co-expression system exhibited optimal activity at a temperature of 35°C, pH 7.5, and the glucose to PPO molar ratio of 1.4:1. In a 30 L bioconversion system the final concentration and productivity of L-PPT reached 79.8 g/L (>99% ee ) and 1440 g/(L·d), respectively. These results will contribute to the industrial biological production of L-PPT.
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INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2025
·
IF 8.5
Qihui Lu,Hui Liao,Zedong Jiang,Yanbing Zhu,Yijuan Han,Lijun Li,Hui Ni,Qingbiao Li
Glycosylation plays a critical role in regulating activity, stability, and correct folding of enzymes. In this study, recombinant Aspergillus niger α-L-rhamnosidase (r-Rha1) was employed to explore the impact of glycosylation in Pichia pastoris on the enzymatic properties and protein folding . β-elimination reaction and deglycosylase treatment assays demonstrated that r-Rha1 undergoes primarily N-glycosylation. The deglycosylated r-Rha1 was prepared in two ways: treating with Endoglycosidase F 1 after expression (referred to as r-Rha1- vitro ), or inhibiting intracellular glycosylation using tunicamycin (referred to as r-Rha1- vivo ). Deglycosylation resulted in a 0.22-fold decrease in activity for r-Rha1- vitro and due to its slower turnover rate, r-Rha1- vivo showed a 0.73-fold decrease in activity. r-Rha1- vitro maintained the similar optimal temperature as r-Rha1, r-Rha1- vivo displayed a 10 °C lower optimal temperature. Compared to the decreased extent of r-Rha1- vitro in t 1/2 at 55 °C, 60 °C, and 65 °C and T m , chemical interferent deglycosylation in vivo showed a more profound impact on r-Rha1. Analyses based on circular dichroism, fluorescence spectroscopy, and differential scanning calorimetry revealed significant changes in the structure and thermodynamic stability of r-Rha1- vivo , accounting for its marked decline in activity and stability. The significant and unpredictable structure changes of r-Rha1- vivo proved the essential role of glycosylation for appropriate folding in P. pastoris .
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Food Bioscience
·
2024
·
IF 4.8
Jiatao Zhang,Bowen Guan,Yuan Zhang,Jian Hu,Tao Sun,Tungalag Dong,Xueyan Yun
An new high-barrier packaging films for meat preservation were obtained by depositing graphene oxide (GO) layer-by-layer (LbL) on poly( l -lactic acid)(PLLA)-poly (butylene itaconate) (PBI) copolymers (PLBI) with chitosan (CS) as bonding layer in this study. The 1 H NMR results revealed that PLBI copolymer was successful synthesized. The introduction of PBI reduced the T g and crystallization capacity of PLLA film, which made the copolymer films exhibit extremely high flexibility (up to 186.1% elongation at break). The film also exhibited high hydrophilicity, which was favorable for the effective attachment of CS. With CS as an intermediate layer, the FTIR and SEM results showed that the GO was evenly distributed onto the PLBI film surface due to the electrostatic interaction. The oxygen, water vapor and UV light barrier properties of PLBI/CS/(GO-CS) x films were improved and increased with the GO-CS layers. The PLBI/CS/(GO-CS) 10 films, with extremely low oxygen permeability coefficients about 0.4 × 10 −8 cm 3 m/m 2 ·h·Pa, delayed the increase in weight loss, hardness, pH, TVB-N, TBARS, and total viable counts of chilled meat during storage. It also decreased the discoloration, and loss of springness and cohesiveness of chilled meat caused by long-term refrigeration. The LbL films even showed better freshness-keeping effect than commercial PA/PE films due to its superior barrier and certain antibacterial properties, suggesting great potential application in fresh meat preservation.
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Science Advances
·
2025
·Vol. 11
·期 3
·
IF 12.5
Yikai Yun,Qing Chang,Jinjian Yan,Yuanyuan Tian,Sijie Jiang,Wenjie Wei,Shaoqun Li,Yuzheng Guo,Jun Yin,Jing Li,Mengyu Chen,Kai Huang,Cheng Li,Rong Zhang
The utilization of low-dimensional perovskites (LDPs) as interlayers on three-dimensional (3D) perovskites has been regarded as an efficient strategy to enhance the performance of perovskite solar cells. Yet, the formation mechanism of LDPs and their impacts on the device performance remain elusive. Herein, we use dimensional engineering to facilitate the controllable growth of 1D and 2D structures on 3D perovskites. The differences of isomeric ligands in electrostatic potential distribution and steric effects for intermolecular forces contribute to different LDPs. The 1D structure facilitates charge transfer with favored channel orientation and energy level alignment. This approach enables perovskite solar modules (PSMs) using 2,2′,7,7′-tetrakis[N,N-di(4-methoxyphenyl)amino]-9,9′-spirobifluorene to achieve an efficiency of 20.20% over 10 by 10 square centimeters (cm2) and 22.05% over 6 by 6 cm2. In particular, a PSM (6 by 6 cm2) using poly[bis(4-phenyl)(2,4,6-trimethylphenyl)amine] maintains an initial efficiency of ~95% after 1000 hours under the rigorous ISOS-L-3 accelerated aging tests, marking a record for the highest stability of n-i-p structure modules.
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JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2024
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IF 5.7
Kai Zhu,Jinghua Wu,Ankai Hu,Zihao Yin,Zhiqiang Hou,Xingqian Ye,Shiguo Chen
A primary challenge of polysaccharide analysis is the need for comprehensive extraction and characterization methods. In this study, mulberry polysaccharides at different maturities were fully extracted through a two-step process involving ethylenediaminetetraacetic acid (EDTA) and sodium hydroxide (NaOH), and their structures were determined by a combination analysis of monosaccharides and glycosidic linkages based on liquid chromatography triple quadrupole mass spectrometry (LC/QqQ-MS). The results indicate mulberry polysaccharides mainly contain highly branched pectic polysaccharides, (1,3,6)-linked glucan, xylan, and xyloglucan, but the content of different portions varies at different maturity stages. HG decreases from 19.12 and 19.14% (green mulberry) to 9.80 and 6.08% (red mulberry) but increases to 17.83 and 11.83% as mulberry transitioned from red to black. In contrast, the contents of glucan showed opposite trends. When mulberry turns red to black, the RG-I arabinan chains decrease from 47.75 and 28.86% to 13.16 and 12.72%, while the galactan side chains increase from 1.18 and 1.91 to 8.3 and 6.49%, xylan and xyloglucan show an increase in content. Overall, the two-step extraction combined with LC/QqQ-MS provides a new strategy for extensive analysis of complex plant polysaccharides.
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INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2025
·
IF 8.5
Jing Wen,Shue Jin,Xue Luo,Chunhong Chen,Huan Liu,Yubao Li,Jidong Li
In cases of deep skin defects, spontaneous tissue regeneration and excessive collagen deposition can result in the formation of proliferative scarring. Salvianolic acid B (SAB) demonstrates promising applications in the treatment of fibrotic diseases, including scarring. This study prepared biodegradable gelatin (Gel) and poly(lactic- co -ε-caprolactone) (PLCL) nanofibrous membranes using electrospinning technology and functionalized them with salvianolic acid B to create fibrous membrane dressings with anti-scarring properties. The resulting Gel/PLCL/SAB (GPS) fibrous membrane exhibited good mechanical properties and biodegradability. In vitro experiments demonstrated that the fibrous membranes of Gel/PLCL containing 0.5 wt% SAB (GPS 0.5 ) exhibited good cytocompatibility, regulated macrophage function and polarization, and inhibited fibrosis-related genes, including CD36, α-SMA, and collagen I. In a mouse model of full-thickness skin defects, the GPS 0.5 fibrous membrane effectively modulated the immune microenvironment and promoted vascular regeneration, thereby accelerating skin healing. Furthermore, the GPS 0.5 fibrous membrane reduced the expression of CD36 and TGF-β1, downregulated collagen I and III, and promoted the regeneration of hair follicles, sebaceous glands, and other appendages, ultimately reducing scar formation. Thus, SAB-loaded Gel/PLCL fibrous membranes hold potential as bifunctional skin dressings that promote wound healing and inhibit fibrosis.
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Analytical Methods
·
2024
·Vol. 16
·期 23
·3720-3731
·
IF 2.7
Rongrong Cao,Songlin Xu,Zhirui Yu,Liang Xu,Zhiqiang Ge,Qianyu Huo,Guoqing Zhu,Bin Qiao
In recent years, there has been a growing interest in the thriving monoclonal antibody (mAb) industry due to the wide utilization of mAbs in clinical therapies. Robust and accurate bioanalytical methods are required to enable fast quantification of mAbs in biological matrices, especially in the context of pharmacokinetics (PKs)/pharmacodynamics (PDs) and therapeutic drug monitoring (TDM) studies. In this investigation, we presented a novel immuno-magnetic capture coupled with a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method designed for the quantification of immunoglobulin G-kappa-based mAbs in biological fluids. The immunoaffinity absorbent for mAb drug purification was meticulously crafted by immobilizing protein L onto monosize, magnetic poly(glycidyl methacrylate) (m-pGMA) beads, synthesized through dispersion polymerization. The microspheres were acquired with an average size of 1.6 μm, and the optimal binding of mAbs from the aqueous mAb solution was determined to be 45.82 mg g−1. The quantification of mAbs in 10 μL serum samples was achieved through affinity purification using m-pGMA@protein L beads (employing rituximab as an internal standard (IS)), on-bead reduction, and rapid tryptic digestion. Remarkably, the entire process, taking less than 2.5 hours, held significant potential for simplifying pretreatment procedures and minimizing analytical time. Furthermore, the developed method underwent validation in accordance with the European Medicines Agency (EMA) guidelines. The assay demonstrated commendable linearity within the 2–400 μg mL−1 range for both daratumumab and pembrolizumab. Intra- and inter-assay coefficients of variation fell within the range of 0.7% to 13.4%, meeting established acceptance criteria. Other validation parameters also conformed to regulatory standards. Ultimately, the efficacy of the method was substantiated in a pharmacokinetic study following a single-dose intravenous administration to mice, underscoring its applicability and reliability in real-world scenarios.
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Frontiers in Pharmacology
·
2024
·
IF 4.4
Xin Yang,Canhui Hong,Tangfei Guan,ChengGui Zhang,Peiyun Xiao,Yongshou Yang,Huai Xiao,Zhengchun He
Ischemic stroke (IS) is a highly lethal type of cardiovascular and cerebrovascular disease. Improving survival rates and promoting recovery in patients with IS pose significant challenges, however, recent research has identified the gut–brain axis as a therapeutic target. In this study, we evaluated the regulatory effect of Periplaneta americana (L.) extract (PAS840), which has established anti-inflammatory, antioxidant, and neuroprotective effects, on the gut microbiota using a rat model of temporary middle cerebral artery occlusion (tMCAO). We evaluated the protective effects of PAS840 on brain damage in IS rats through TTC (triphenyltetrazolium chloride), Nissl staining, and pathological section analysis. Additionally, we investigated the impact of PAS840 on the gut microbiota and metabolites using 16S rRNA sequencing, untargeted metabolomics of gut contents, and transcriptomics analyses of brain tissues to explore its mechanism of action. PAS840 intervention resulted in significant changes in the gut microbiota, including an increase in the abundance of probiotic flora, decrease in the abundance of harmful flora, and significant changes in metabolite profiles. It also attenuated brain damage, decreased platelet activity, inhibited oxidative stress and genes related to inflammation, and improved neurological function in rats. These findings suggest that PAS840 has preventive and therapeutic effects against IS via the gut–brain axis by regulating the gut microbiota and related metabolites. Accordingly, PAS840 is a candidate therapeutic drug for further research.
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CARBOHYDRATE POLYMERS
·
2024
·
IF 10.7
Shuting Peng,Sen Niu,Qin Gao,Ruiyuan Song,Zhengxiao Wang,Ziyun Luo,Xi Zhang,Xiaofei Qin
The biggest obstacle to treating wound healing continues to be the production of simple, inexpensive wound dressings that satisfy the demands associated with full process of repair at the same time. Herein, a series of injectable composite hydrogels were successfully prepared by a one-pot method by utilizing the Schiff base reaction as well as hydrogen bonding forces between hydroxypropyl chitosan (HCS), ε-poly- l -lysine (EPL), and 2,3,4-trihydroxybenzaldehyde (TBA), and multiple cross-links formed by the reversible coordination between iron (III) and pyrogallol moieties. Notably, hydrogel exhibits excellent physicochemical properties, including injectability, self-healing, water retention, and adhesion, which enable to fill irregular wounds for a long period, providing a suitable moist environment for wound healing. Interestingly, the excellent hemostatic properties of the hydrogel can quickly stop bleeding and avoid the serious sequelae of massive blood loss in acute trauma. Moreover, the powerful antimicrobial and antioxidant properties also protect against bacterial infections and reduce inflammation at the wound site, thus promoting healing at all stages of the wound. The study of biohydrogel with multifunctional integration of wound treatment and smart medical treatment is clarified by this line of research.
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MOLECULAR NUTRITION & FOOD RESEARCH
·
2024
·
IF 4.5
Qiuming Liu,Qingyu Ma,Jiao Li,Liping Sun,Yongliang Zhuang
Scope Type 2 diabetes mellitus (T2DM) is a metabolic disease with a major global public health effect. Rambutan peel polyphenols (RPPs) have been reported to exert hypoglycemic activity. However, few studies have been explored from the viewpoint of gut microbiota and its metabolites. Methods and results RPPs are administered by gavage to a mice model of T2DM established by using a high-fat diet combined with streptozotocin. It finds that RPPs treatment alleviates hyperglycemia symptoms by improving glucolipid metabolism and liver function. Immunohistochemistry indicates that the antihyperglycemic effect of RPPs is regulated by the IRS-1/PI3K/AKT/GSK3β signaling pathway. RPPs treatment remodels the structure of gut microbiota ( Odoribacter , Lachnospiraceae_NK4A136_group , Lactobacillus , Turicibacter , Erysipelatoclostridium , and Tuzzerella ) and enriches the metabolites (RPPs-derived urolithins, short-chain fatty acids, dehydrocholic acid, (+)-catechin, dihydroberberine, pterostilbene, and artesunate) associated with diabetes regulation in T2DM mice. The effects of RPPs in ameliorating glycolipid metabolism disorders are correlated with differential gut microbiota and metabolites. Conclusion The gut microbiota and its metabolites are key targets for the hypoglycemic effects of RPPs.
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Foods
·
2024
·Vol. 13
·期 11
·1615
·
IF 4.7
Yuting Fu,Shuai Chen,Xinjun Wang,Lu Wang,Zexin Wang,Yanfei Cheng,Yuyi Liu,Lin Zhang,Sixin Liu,Jiamu Kang,Congfa Li
White pepper, used both as a seasoning in people’s daily diets and as a medicinal herb, is typically produced by removing the pericarp of green pepper through the retting process. However, the mechanism of the retting process for peeling remains unclear. Therefore, this study aimed to investigate the changes in physicochemical factors, microbial community succession effects, and metabolites of the pepper pericarp during the pepper peeling process. The findings indicated that pre-treatment involving physical friction before the retting process effectively reduced the production time of white pepper. During the retting process, the pectinase activity increased, leading to a decrease in the pectin content in the pepper pericarp. There was a significant correlation observed between the changes in pH, pectin content, and peeling rate and the Shannon diversity index of bacteria and fungi.Prevotella,Lactococcus, andCandidawere the dominant microbial genera during the retting. The functional predictions suggested that the monosaccharides degraded from the pepper pericarp could have been utilized by microbes through sugar metabolism pathways. Metabolomic analysis showed that the metabolic pathways of carbohydrates and amino acids were the main pathways altered during the pepper peeling process. The verification experiment demonstrated that the degradation of pectin into galacturonic acid by polygalacturonase was identified as the key enzyme in shortening the pepper peeling time. The structure of the pepper pericarp collapsed after losing the support of pectin, as revealed by scanning electron microscopy. These results suggest that the decomposition of the pepper pericarp was driven by key microbiota. The succession of microbial communities was influenced by the metabolites of the pepper pericarp during retting. These findings provide new insights into the retting process and serve as an important reference for the industrial production of white pepper.
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PROCESS SAFETY AND ENVIRONMENTAL PROTECTION
·
2025
·
IF 7.8
Fenyan Chen,Wenqing Zhou,Lihua Qi,Jingjing Yang,Zhiman Li,Xiaoge Zhang,Yalan Zhu,Chunfeng Guan
The excessive accumulation of dimethyl phthalate (DMP) in soil exerts tremendous pressure on soil ecosystems and human health. This study explored the feasibility of using bacterial quorum sensing signal molecules, N -acyl-homoserine lactones (AHLs), to enhance phytoremediation of DMP contaminated soil. The effects of N -butyryl- L -homoserine lactone (C 4 -HSL) on soybean ( Glycine max L.) physiology and phytoremediation efficiency were assessed. Results indicated that C 4 -HSL significantly promoted the efficiency of soybean in remediating DMP contaminated soil, achieving an 87.40 % DMP removal efficiency after 28 d cultivation. Applying C 4 -HSL significantly enhanced soybean photosynthetic by the potential promotion of chlorophyll synthesis and bolstered the antioxidant with a notable reduction in malondialdehyde content. The presence of C 4 -HSL also stimulated plant growth and improved soil enzymatic activities, likely aiding in nutrient cycling and pollutant degradation in soil. Moreover, C 4 -HSL modified the bacterial community, increasing the relative abundance of bacteria related to DMP degradation (Proteobacteria, Actinobacteria) and plant growth promotion (Micromonosporales, Sphingomonadaceae). In general, this study proposed that AHLs-assisted phytoremediation offers a promising, eco-friendly strategy for DMP remediation. This approach provides economic and ecological benefits while reducing damage to soybeans and lays the groundwork for practical applications in agriculture.
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