Applied Biochemistry and Biotechnology最新文献

{"title":"Refinement of Synthetization Parameters for High Laccase-Like Activity of Imidazole-Copper (II) Nitrate Trihydrate Nanozyme Towards an Efficient Biomimetic Nanozyme.","authors":"Nur Aizura Mat Alewi, Roshanida A Rahman, Rosli Md Illias, Nardiah Rizwana Jaafar, Noor Hidayah Abd Rahman, Bee Jie Chia, Hui Lun Soo, Ariadne L Juwono, Munawar Khalil, Noverra M Nizardo","doi":"10.1007/s12010-025-05229-w","DOIUrl":"https://doi.org/10.1007/s12010-025-05229-w","url":null,"abstract":"<p><p>Laccase's industrial application is hindered by its sensitivity and low stability to extreme conditions. To overcome these limitations, the development of biomimetic nanozymes is gaining momentum. Nevertheless, developing multifunctional nanozymes with high laccase-like activity poses several challenges. This study focused on optimizing the synthesis of imidazole-copper (II) nitrate trihydrate (I-Cu) nanozymes and characterizing its physicochemical properties. Key synthesis parameters (precursor amount, incubation time, and oven temperature) were optimized. I-Cu nanozymes were synthesized in a Teflon-lined autoclave via water-induced precipitation of Cu²⁺ and imidazole, mimicking the N-Cu coordination found in laccase's active sites. Initial screenings revealed the superior catalytic activity of I-Cu nanozymes synthesized using methanol compared to ethanol, and a smaller nano-scale size than laccase. FTIR analysis confirmed the presence of similar chemical components as laccase (C₄₄H₆₉N₁₁O₂₀), verifying I-Cu nanozyme's capability to degrade phenolic compounds, and imidazole did not decompose throughout the synthesis process. The optimized I-Cu nanozyme demonstrated higher catalytic activity (6.569 UA), oxidation efficiency (V_max of 0.00893 mM/min and K_m of 2.4020 mM), and greater stability under varying pH, temperature, and storage conditions, compared to laccase. Conclusively, the optimized I-Cu nanozyme, with a 6.00-fold increase in catalytic activity compared to previous studies, as well as 1.69-fold higher K_m, and 2.08-fold higher V_max compared to laccase, shows promise as a wastewater treatment alternative. Its enhanced performance, achieved with fewer precursors through synthesis optimization, highlights the potential of lesser-known biomimetic nanozymes and underscores the importance of refining the synthesis parameters.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143727054","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mannanase from Lactic Acid Bacteria Pediococcus acidilactici: Production Optimization, Purification, and Application in Juice Clarification.","authors":"Jingjun Shi, Xi Chen, Ruoxi Yang, Dan Zhao","doi":"10.1007/s12010-025-05231-2","DOIUrl":"https://doi.org/10.1007/s12010-025-05231-2","url":null,"abstract":"<p><p>Pediococcus acidilactici, a generally regarded as safe (GRAS) lactic acid bacteria species, is a desirable microbial mannanase source due to its high bio-safety. P. acidilactici HDM2 isolated from dragon fruit culture was identified according to morphological, physiological, and biochemical characteristics as well as preserved nucleic acid (16S rDNA and rpoA) sequencing. A two-step response surface methodology (RSM) experiment subsequently screened out and then predicted the optimal levels of two factors which significantly affected P. acidilactici HDM2's mannanase production. The maximal enzyme activity reached 97.9 U/mL under the condition of glucose 17.5 g/L and initial pH 4.6. RSM optimization resulted in a 39.3% increase of mannanase yield which was verified by six repeated trials and consequent t-test (P < 0.05). The enhancement of mannanase production was also proved by congo-red dyeing on KGM (konjac glucomannan)-MRS (de Man, Rogosa, and Sharpe) agar. Diameter of clearance zone enlarged from 2.7 to 4.5 cm through RSM. The purified P. acidilactici HDM2 mannanase with a molecular weight of 30 KDa was then applied in fruit juice clarification process. The enzyme exhibited significantly higher clarifying efficiency than commercial mannanase in both yield and clarity for six out of eight fruits, i.e., orange, peach, grape, pear, kiwi, and grapefruit. These results promised the application of P. acidilactici HDM2's mannanase in various especially food-level fields.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143727050","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Review on the Over-liming Detoxification of Lignocellulosic Biomass Prehydrolysate for Bioethanol Production.","authors":"Kar Fee Chong, Yi Lu, Yuke Han, Yuli Shen, Suchithra Thangalazhy-Gopakumar, Suan Shi, Lujia Han","doi":"10.1007/s12010-025-05212-5","DOIUrl":"https://doi.org/10.1007/s12010-025-05212-5","url":null,"abstract":"<p><p>There is an increasing interest in utilizing biomass for biofuel production to mitigate the negative environmental impact caused by fossil fuels. The production of ethanol from lignocellulosic biomass necessitates a pre-treatment process to break down the biomass into fermentable sugars. However, the use of high temperatures and chemical concentrations during pre-treatment leads to the formation of inhibitors, adversely impacting ethanol production yields. Detoxification, the process of removing inhibitors by physical adsorption or by converting them into less harmful compounds, is crucial in preventing inhibition during fermentation. Over-liming detoxification is one of the most employed methods for ethanol production due to its high efficiency, easy operation, and low cost. In addition to the types of alkali used, factors like pH value, temperature, and incubation time also play a crucial role in its effectiveness. Therefore, determining optimal detoxification conditions and selecting suitable alkalis are essential to the successful removal of inhibitors from biomass hydrolysate, ultimately enhancing ethanol production. Suitable microorganisms used during fermentation also have a substantial impact on ethanol production. This review paper summarizes the effect of the alkali detoxification method on the ethanol fermentation of lignocellulosic biomass. The limitations and problems of over-liming detoxification and potential methods that could enhance the over-liming are also discussed.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143727473","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Astragaloside IV Relieves Mitochondrial Oxidative Stress Damage and Dysfunction in Diabetic Mice Endothelial Progenitor Cells by Regulating the GSK-3β/Nrf2 Axis.","authors":"Xiaoling Zou, Xiangnan Liu, Wenjing Qu, Xi Zhang, Yixian Zou, Xiangdong Lin, Wenxiao Hu, Ruchun Gao, Youyuan He, Siyuan Zhou, Yuxuan Huang, Wu Xiong","doi":"10.1007/s12010-025-05211-6","DOIUrl":"https://doi.org/10.1007/s12010-025-05211-6","url":null,"abstract":"<p><p>Dysregulation of mitochondrial activity is a major cause of diabetes mellitus (DM) and its complications. Astragaloside IV, a natural herbal product, possesses protective properties against DM. This study aimed to evaluate how astragaloside IV affects oxidative stress and mitochondrial function in endothelial progenitor cells (EPCs) and elucidate the underlying mechanisms. A high glucose (HG)-induced human EPC (hEPC) model and a streptozotocin (STZ)-induced DM mouse model were established to investigate the effects of astragaloside IV on EPC function and wound healing in the context of DM. In HG-exposed hEPCs, astragaloside IV reduced apoptosis and increased cell viability and tube formation (P < 0.05). In STZ-induced DM mice, astragaloside IV promoted wound healing and increased the expression of the endothelial marker CD31 (P < 0.05) in wound tissues. In addition, the regulation of oxidative damage and mitochondrial dysfunction by astragaloside IV was investigated. We found that astragaloside IV attenuated mitochondrial damage, decreased ROS and mtROS levels (P < 0.05), decreased MDA activity and enhanced SOD activity (P < 0.05), and downregulated DPR1 levels and upregulated MFN1, MFN2, and OPA1 levels (P < 0.05). Mechanistically, the potential involvement of GSK-3β/Nrf2 was investigated by molecular docking and intervention with the GSK-3β activator sodium nitroprusside (SNP). Astragaloside IV was confirmed to dock with GSK-3β, and it increased the phosphorylation of GSK-3β (P < 0.05) and the expression of Nrf2 as well as its downstream factors HO-1 and NQO1 (P < 0.05). SNP reversed the protective effects of astragaloside IV. These results indicated that astragaloside IV attenuated HG- and STZ-induced injury through the GSK-3β/Nrf2 pathway. These results revealed that astragaloside IV may have the potential to be an active component for protection against DM and its complications.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143707827","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Role and Validation of Lactylation-Related Gene Markers in Postmenopausal Osteoporosis.","authors":"Fuzhu Tan, Xinmei Cui, Shujun Ren, Yu Zhang","doi":"10.1007/s12010-025-05216-1","DOIUrl":"https://doi.org/10.1007/s12010-025-05216-1","url":null,"abstract":"&lt;p&gt;&lt;strong&gt;Background: &lt;/strong&gt;Osteoporosis (OP) is a systemic bone disease characterized by bone loss, disrupted bone structure, and increased susceptibility to fractures. Postmenopausal osteoporosis (PMOP) refers to OP that occurs in women during the late postmenopausal period, with the main cause being a decrease in estrogen levels. Lactylation, as a glycation modification, may affect bone cell function in PMOP. However, its specific role in the development of PMOP remains unclear.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Methods: &lt;/strong&gt;In this study, we collected single-cell RNA sequencing (scRNA-seq) data and transcriptome data of PMOP. The scRNA-seq data were processed using the \"Seurat\" package, including cell filtering, normalization, dimensionality reduction, and clustering. The cell types were annotated using the \"singleR\" package. Based on lactylation-related genes (LRGs), all cells were divided into high- and low-expression cell groups. Differences in signaling pathways, developmental trajectories, and transcription factor activity between the two cell groups were explored using the \"fgsea,\" \"monocle,\" and \"DoRothEA\" packages, respectively. Mendelian randomization (MR) analysis was performed to identify genes with significant differential expression between the two cell groups that are causally related to PMOP. The differentially expressed genes (DEGs) between the high- and low-expression cell groups were selected using the \"limma\" package, and the intersection with DEGs was taken. A diagnostic model for PMOP was constructed using multiple machine learning algorithms and their combinations based on the intersection genes. Immune infiltration analysis was performed on the transcriptome data using the ssGSEA algorithm. Finally, a column line plot model of PMOP was constructed based on diagnostic genes.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Results: &lt;/strong&gt;After annotating the cell types in the scRNA-seq data, a total of 11 cell types were obtained, including neutrophils, tissue stem cells, monocyte, macrophage, erythroblast, myelocyte, Pre-B cell CD34-, BM, T cells, B cell, and Pro-B cell CD34 + . The high- and low-expression cell groups divided based on the expression levels of LRGs showed significant differences in signaling pathways, developmental trajectories, and transcription factor activity. The MR analysis identified RPS10 and RPL12 as risk factors causally related to PMOP. A diagnostic model for PMOP was constructed based on the transcriptome data and the intersection DEGs between the high- and low-expression cell groups. The model achieved AUCs of 0.961, 0.730, and 0.9 in the training set and two testing sets, respectively, indicating high predictive accuracy. Additionally, eight diagnostic genes, including S100A9, ARHGEF10, RPL30, ANGPT1, RPL18, LAMB1, RBMS3, and RPL27A, were identified. The column line plot model constructed based on these diagnostic genes also showed high AUCs and clinical utility.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Conclusion: &lt;/strong&gt;This study revealed the important r","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143707830","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Phytochemical Investigation, Antibacterial and Antioxidant Activity Determination of Leaf Extracts of Cynoglossum coeruleum (Shimgigit).","authors":"Tsegaye Atnaf Chekol, Belete Bedemo Beyene, Addisu Taddele Teshale, Shewa Getachew Mamo","doi":"10.1007/s12010-025-05226-z","DOIUrl":"https://doi.org/10.1007/s12010-025-05226-z","url":null,"abstract":"<p><p>The leaf of Cynoglossum coeruleum is traditionally used in tropical medicine for treating various ailments. This study analyzed the phytochemicals, flavonoid and phenolic content, antioxidant capacity, and antibacterial activity of its leaf extracts. Phytochemical tests revealed the presence of alkaloids, flavonoids, phenols, steroids, terpenoids, and saponins. The total flavonoid content in methanol, ethyl acetate, and petroleum ether extracts was 74.66 mgQE/g, 67.34 mgQE/g, and 15.39 mgQE/g, respectively. Total phenolic content was 61.93 mg GAE/g, 25.61 mg GAE/g, and 17.72 mg GAE/g. The extracts showed significant antibacterial activity against Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, and Staphylococcus epidermidis at higher concentrations.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-03-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143699293","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Potassium Hydroxide-Mediated Pretreatment of Sugarcane Bagasse: High-Efficiency Enzymatic Hydrolysis and Excellent Sugar Recovery.","authors":"Zhaoyu Wang, Xuewen Chi, Hanwen Feng, Xiaoya Yao, Yanhong Bi, Rongling Yang, Bin Li","doi":"10.1007/s12010-025-05225-0","DOIUrl":"https://doi.org/10.1007/s12010-025-05225-0","url":null,"abstract":"<p><p>Sugarcane bagasse, an important by-product of the sugar industry, is the fibrous residue remaining from sugarcane stalks following the extraction of sugarcane juice. In this study, the efficacy of potassium hydroxide (KOH) pretreatment on sugarcane bagasse under mild conditions is examined, and its effectiveness was assessed through enzymatic saccharification at various solid loadings. After pretreatment with a KOH dosage of 15 wt.%, a solid-liquid ratio of 1:10, and reaction time at 90 °C for 2 h, enzymatic saccharification experiments were performed to observe the change rule for the sugar recovery yield. The results showed that the glucose concentration achieved 47.6 ± 1.1 g/L under optimal conditions. With a specified solid loading of 5% and an enzyme dosage of 20 FPU cellulase/g-substrate, the total sugar yield reached as high as 81.6 ± 1.1%. Additionally, the entire fermentable sugar production process was clean and sustainable, which is crucial for converting lignocelluloses to advanced bioenergy or chemicals.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-03-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143699383","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Biochemical Characterization of a GH11 Xylanase from Xylanase-Producing Trichoderma citrinoviride.","authors":"Beom Soo Kim, Bo Min Jeong, Dongeon Kim, Soo Rin Kim, In Jung Kim","doi":"10.1007/s12010-025-05213-4","DOIUrl":"https://doi.org/10.1007/s12010-025-05213-4","url":null,"abstract":"<p><p>Xylan, a prevalent component of lignocellulose, ranks as the second most abundant carbohydrate in nature. Endo-1,4-xylanase, pivotal for its ability to cleave β-1,4-glycosidic linkages within xylan, is crucial for various applications in the food/feed processing, biofuel production, and paper/pulp industries. Although Trichoderma citrinoviride is renowned for its robust xylan-degrading capacity, the biochemical properties of xylanases derived from T. citrinoviride remain largely uncharacterized. Therefore, this study was conducted to explore the biochemical characteristics of a glycoside hydrolase family 11 xylanase derived from T. citrinoviride (TciGH11). This newly identified enzyme efficiently hydrolyzed beechwood xylan into xylooligosaccharides, exhibiting optimal activity at pH 4.5 and 50 °C, with a specific activity of 1801.5 U mg^-1 against beechwood xylan. Kinetic analysis revealed a Michaelis constant (K_m), turnover number (k_cat), and catalytic efficiency (k_cat/K_m) of 3.82 mg ml^-1, 977.8 s^-1, and 256.0 ml mg^-1 s^-1, respectively. Structural modeling analysis demonstrated that the non-conserved residues in TciGH11, specifically Glu162 and Ser165, affected the substrate binding properties, potentially resulting in different enzymatic activity compared to that of other xylanases from the Tricoderma genus. Overall, these results could be instrumental in advancing the utilization of lignocellulosic biomass, thereby supporting sustainable bioprocessing of lignocellulosic biomass.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-03-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143699337","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"6-Gingerol Inhibits Ferroptosis in Endothelial Cells in Atherosclerosis by Activating the NRF2/HO-1 Pathway.","authors":"Shuai Wang, Xiaoliang Song, Hui Gao, Yi Zhang, Xin Zhou, Fengrong Wang","doi":"10.1007/s12010-025-05214-3","DOIUrl":"https://doi.org/10.1007/s12010-025-05214-3","url":null,"abstract":"<p><p>Targeting endothelial cell ferroptosis is a potential approach for the treatment of atherosclerosis (AS). 6-Gingerol (6-Gin) is an active substance in ginger that is beneficial for improving AS. We conducted this study to explore whether 6-Gin mediated AS progression by regulating ferroptosis of endothelial cells. ApoE-/- mice were fed a high-fat diet to establish AS mouse model. Additionally, oxidized low-density lipoprotein (ox-LDL) was used to treat human umbilical vein endothelial cells (HUVECs) to generate injured cell model. Ferroptosis was evaluated by propidium iodide staining assay, western blot, and detecting iron, glutathione, malonaldehyde, and reactive oxygen species levels. The results showed that ox-LDL inhibited the proliferation and induced inflammation and ferroptosis of HUVECs, which was reversed by 6-Gin treatment. Moreover, 6-Gin upregulated HO-1 and NQO1 levels and promoted nuclear translocation of NRF2 in ox-LDL-treated HUVECs. ATRA, an NRF2 inhibitor, abrogated the promotion of proliferation and the inhibition of inflammation and ferroptosis induced by 6-Gin. Additionally, 6-Gin alleviated AS and suppressed ferroptosis in vivo. In conclusion, 6-Gin inhibited endothelial cell ferroptosis by inactivating the NRF2/HO-1 pathway, thereby improving abnormal lipid metabolism in AS mice. These findings suggest that 6-Gin may be a novel therapeutic drug for AS.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-03-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143690689","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Constructing Genetically Engineered Escherichia coli for De Novo Production of L-Threo-3-Hydroxyaspartic Acid.","authors":"Jing Guo, Jiayi Cui, Mingyue Xun, Wencheng Zhang, Mo Xian, Rubing Zhang","doi":"10.1007/s12010-025-05224-1","DOIUrl":"https://doi.org/10.1007/s12010-025-05224-1","url":null,"abstract":"<p><p>L-threo-3-hydroxyaspartic acid (L-THA) is a non-proteinogenic amino acid that has garnered significant attention due to its diverse biological activities. However, the synthesis of L-THA through enzymatic and whole-cell catalysis requires the expensive substrate L-aspartic acid or L-asparagine, and co-substrate α-ketoglutarate, which limits their large-scale application. Here, this is the first report of engineering E. coli as a cell factory for de novo production of L-THA from glucose by fermentation. Firstly, the asnO gene encoding asparagine hydroxylases from Streptomyces coelicolor was heterologously expressed in E. coli to yield the L-THA producing strain. The formation and configuration of L-THA were characterized by LC-MS and HPLC after FDAA derivatization. Secondly, the pathway genes aspC and asnB, which encode aspartate aminotransferase and asparagine synthase, respectively, were overexpressed to enhance L-THA titer from 49.9 to 90.84 mg/L. Thirdly, the efforts were made to improve the key precursor L-aspartic acid pool by overexpressing the aspartase encoding gene aspA and knocking out aspartate kinase (AK) III encoding gene lysC. The best strain CC03 was obtained and L-THA titer reached 278.3 mg/L in a shake flask, representing an approximately 5.6-fold increase compared to the original strain. Ultimately, 2.87 g/L L-THA was obtained after 32 h fed-batch fermentation. This research underscores the potential use of E. coli fermentation as a feasible platform for de novo biosynthesis of L-THA from glucose, which is amenable to industrial application.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-03-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143690641","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}