Applied Biochemistry and Biotechnology最新文献

{"title":"A Readily Degradable Polyvinyl Alcohol/Starch Membrane Containing Catechol-Grafted Microcrystalline Cellulose for Preserving Fruit Freshness.","authors":"Jiangfei Lou, Aixue Dong, Lingling Hu","doi":"10.1007/s12010-025-05321-1","DOIUrl":"https://doi.org/10.1007/s12010-025-05321-1","url":null,"abstract":"<p><p>Copolymer membranes based on polyvinyl alcohol (PVA) and starch suffer from low mechanical strength and poor fresh-produce preservation and degradation performance. Adding functional cellulose or derivatives to membranes can improve these properties. Herein, free radical grafting polymerization between microcrystalline cellulose (MCC) and catechol (CA) was performed using a horseradish peroxidase (HRP)/acetylacetone/H₂O₂ system, and the product was incorporated into a PVA/Starch membrane to improve its performance. The MCC-CA polymer was structurally characterized by FTIR, ¹H NMR, ¹³C NMR, XRD, and HPLC methods. The covalent binding mechanism during the free radical polymerization process was analyzed and optimized based on HRP enzyme dosage, H₂O₂ dosage, CA dosage, reaction temperature, and reaction duration. The mechanical properties, antioxidant properties, degradation properties, and freshness properties of the Starch/MCC-CA membrane were explored. The results indicated MCC and CA were graft polymers between free radicals; the Starch/MCC-CA membrane had a significant improvement in degradation, free radical scavenging properties, and mechanical properties, with the freshness of bananas maintained for > 10 days, superior to existing commercially available PVA/Starch preservation membranes.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-06-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144525861","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":"Transcriptomic Analysis of Lipid Accumulation in Mucor circinelloides WJ11 Under Submerged and Solid-State Fermentation.","authors":"Bushra Iqbal, Muhammad Tariq Saeed, Tahira Naz, Hassan Mohamed, Wenrui Dang, Asmaa S Ramadan, Adel Eltoukhy, Huaiyuan Zhang, Yuanda Song","doi":"10.1007/s12010-025-05318-w","DOIUrl":"https://doi.org/10.1007/s12010-025-05318-w","url":null,"abstract":"<p><p>The oleaginous filamentous fungus Mucor circinelloides was extensively investigated for bio-oil production using submerged fermentation (SmF) and solid-state fermentation (SSF) techniques. SSF fermentation may offer advantages over SmF, largely due to the fungus's unique filamentous structure and the lower costs associated with this strategy. In this study, M. circinelloides strain WJ11 exhibited lower fatty acid (FA) content, reaching up to 12% in SSF compared with 24.1% in SmF after 120 h. Furthermore, transcriptomic analysis observed that 7615 genes were differentially expressed between the two fermentation methods, with 3891 genes upregulated and 3724 downregulated in SSF. Notably, genes encoding triglyceride-related synthases were significantly downregulated in WJ11. The mRNA levels of ATP-citrate lyase were also reduced, indicating a decreased supply of acetyl-CoA for fatty acid biosynthesis in SSF. Additionally, isoleucine and leucine metabolism do not contribute to acetyl-CoA production, further hindering FA synthesis. For NADPH production, which is crucial for FA biosynthesis, the pentose phosphate pathway (PPP) was significantly downregulated, and the activity of NADP⁺-dependent malic enzyme was also reduced under SSF. Additionally, the downregulation of key genes contributes to the glycerolipid pathway and FA biosynthesis during SSF, confirming a strong correlation between these pathways and lipid biosynthesis in WJ11.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-06-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144525863","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":"Abnormal Gut Microbiota is Associated with Depressive-like Behavior in the Rat Model of Autoimmune Prostatitis.","authors":"Yihan Wang, Xin Zhu, Yandong He, Wenlong Lu, Zhong Wang, Feng Liu","doi":"10.1007/s12010-025-05294-1","DOIUrl":"https://doi.org/10.1007/s12010-025-05294-1","url":null,"abstract":"<p><p>Many patients with chronic prostatitis (CP)/chronic pelvic pain syndrome (CPPS) also experience depression, although the pathogenesis was unclear. Some studies have shown that gut microbiota disorder might be related to both prostatitis and depression. Therefore, we investigated the alteration of the gut microbiota in autoimmune CP/CPPS complicated with depression in a rat model. The rat model of autoimmune CP with comorbid depressive-like behavior was constructed by experimental autoimmune prostatitis (EAP) and chronic unpredictable mild stress (CUMS). Then, enzyme linked immunosorbent assay (ELISA) was performed to determine the concentration of inflammatory cytokines. The depression-like behaviors in rats were also detected using the open field test (OFT), forced swimming test (FST), and sucrose preference test (SPT). The composition of the gut microbiota and the microbial profile in rats were measured by bioinformatics analysis of the 16S rRNA gene sequence. The level of interleukin (IL)-1β, IL-6, tumor necrosis factor α (TNF-α), and depression-like behavior in rats showed that the CP/CPPS complicated with depression model was established successfully. The results of the 16S rRNA sequence analysis showed disordered abundance, uniformity, and composition of gut microbiota in EAP + CUMS rats. Over 60 gut microbial metabolic pathways were predicted to be disordered among the four groups of rats. Additionally, metabolomics profiling indicated significant differences in EAP + CUMS rats compared to other groups, including reduced PWY-922, PWY-5198 and PWY-7159, and increased PWY0-321 and PWY-6629, which were predicted to be altered by Lactobacillus, Shigella, Blautia, Clostridiales, Comamonadaceae and Rhodospirillales. Our results have shown that gut microbiota disorders are associated with depressive-like behavior in experimental prostatitis. Our findings concerning gut microbiota provide new insights in understanding prostatitis-associated depression.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144504404","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":"Genetically Engineered Yeast for Enhanced Biodegradation of Β-lactam Antibiotics.","authors":"Carolin Pohl, Cindy Rau, Linda Schuster, Uta Gutbier, Stephan Beil, Katrin Lehmann, Hilmar Börnick, Kai Ostermann, Stefan Stolte","doi":"10.1007/s12010-025-05291-4","DOIUrl":"https://doi.org/10.1007/s12010-025-05291-4","url":null,"abstract":"<p><p>High environmental concentrations of pharmaceuticals, particularly antibiotics, have been observed worldwide, while antibiotic concentrations in the ng/L to µg/L range may adversely affect biota and contribute to the formation of antibiotic resistance. The underlying causes of high environmental concentrations include, in addition to the low rate of antibiotic' metabolization and the high rate of usage, especially of β-lactam antibiotics, the insufficient removal by conventional wastewater treatment methods. Consequently, alternative methods need to be developed to remove antibiotics from wastewater-one possibility is the use of enzymes. In this study, the enzyme β-lactamase was secreted by a genetically modified yeast (Saccharomyces cerevisiae) upon recognition of a pheromone (α-factor) as inducer to enable the degradation of ampicillin. This represents a crucial step on the road to a sensor-actuator system, allowing for the development of an intelligent removal system that can react to the presence of antibiotics. Ampicillin and its transformation products were studied by LC-MS/MS measurements using a carbamoyl functionalized column under hydrophilic interaction liquid chromatography (HILIC) conditions, which allowed detection of ampicillin at concentrations of 2.43 nM. The dependence of ampicillin degradation on α-factor concentration and the cultivation time of the yeast was demonstrated, resulting in higher degradation rate with higher α-factor concentrations and longer yeast cultivation times. Over 90% of 10 µM ampicillin was degraded within 0.5 h using 250 nM α-factor and a cultivation period of 24 h. Finally, the transferability to other β-lactam antibiotics was investigated, resulting in complete degradation of amoxicillin, penicillin G and piperacillin within 24 h.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144504405","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":"Green Synthesis of Tin Oxide (SnO₂) Nanoparticles Using Ginger Extracts for Photocatalytic Degradation of Organic Dyes in Wastewater.","authors":"Yuvana Sivarajan, Khairul Anwar Ishak, Jamilah Syafawati Yaacob","doi":"10.1007/s12010-025-05293-2","DOIUrl":"https://doi.org/10.1007/s12010-025-05293-2","url":null,"abstract":"<p><p>One of the main issues that has been receiving great attention globally is the pollution of water resources with dyes, as the chemicals present in them are carcinogenic and harmful to aquatic organisms and humans. Metal oxide nanoparticles synthesized using plant extracts have recently emerged as the best option to address this problem as they provide an eco-friendly and inexpensive approach for a clean water supply. This study describes the photocatalytic degradation of four different types of dyes-methylene blue (MB), rhodamine B (RB), Eriochrome black-T (EB), and methyl orange (MO) using tin oxide nanoparticles (SnO₂ NPs) synthesized with ginger (Zingiber officinale) extract. From the results, the SnO₂ NPs play a remarkable role in degrading all the dyes even though they require a longer time to reach maximum degradation. This study also describes the biosynthetic preparation of SnO₂ NPs, which have a  tetragonal structure, prepared using the optimum parameters of 0.02 g mL^-1 of ginger extract, 0.5 M of tin (II) chloride dihydrate (SnCl₂.H₂O), and reaction pH of 5. SnO₂ NPs were characterized using powder X-ray diffraction (XRD) to evaluate their crystallinity and composition, Fourier transform infrared (FTIR) to identify the changes in functional groups of SnO₂ NPs, and ultraviolet-visible (UV-Vis) spectroscopy to analyze the optical properties of SnO₂ NPs through band gap calculation. The photocatalytic activity of SnO₂ NPs was then studied through their efficiency in degrading the four dyes. A toxicity test was also carried out in this study using brine shrimp (Artemia salina) to ensure the safety of the synthesized SnO₂ NPs made with ginger extract for use in aquatic environments to degrade pollutants. The positive results obtained indicate their potential as a good photocatalyst for water remediation.</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144504406","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":"METTL14/YTHDF2 m6A Axis Protects Against M2 Macrophage Polarization in Endometriosis by Regulating KLF4 Stability","authors":"Li Li,&nbsp;Xiaoyan Wang,&nbsp;You Wu,&nbsp;Yi Liu,&nbsp;Mingzhe Zhang","doi":"10.1007/s12010-025-05290-5","DOIUrl":"10.1007/s12010-025-05290-5","url":null,"abstract":"<div><p>Endometriosis (EMs) is a chronic inflammatory disorder featured by infertility and pain. The role of N6-methyladenosine (m6A) in EMs has been evidenced. This study investigated the molecular mechanism of <i>METTL14</i>-m6A-<i>KLF4</i> modulating macrophage polarization in EMs. RT-qPCR assay was conducted to test <i>METTL14</i> levels in tissues and cells and the relative mRNA levels of M2 (<i>Arg-1</i>, <i>Fizz1</i>) and M1 (<i>iNOS</i>, <i>IL-1β</i>) factors in the supernatant after co-culture of macrophages with normal endometrial stromal cells (nESCs) or ectopic endometrial stromal cells (eESCs). <i>CD206</i> and <i>CD86</i> expression, as well as <i>Arg-1</i>, IL-10, and IL-4 levels, were assessed. Meanwhile, the relationship between <i>METTL14</i> and the m6A modification of <i>KLF4</i> was analyzed. Additionally, the effect of <i>KLF4</i>-activated M2 macrophages on in vitro ESC progression was observed. Cellular and tissue <i>METTL14</i> was under-expressed in EMs. <i>METTL14</i> expression might be related to macrophage M2 polarization. Co-culture of eESCs overexpressing <i>METTL14</i> and macrophages downregulated <i>Arg-1</i>, <i>Fizz1</i>, <i>CD206</i>, IL-10, and IL-4 levels. Mechanistically, <i>METTL14</i> could mediate <i>KLF4</i> m6A modification through the m6A reading protein <i>YTHDF2</i>. <i>KLF4</i> overexpression could nullify <i>METTL14</i> re-expression-repressed M2 macrophage polarization. In addition, <i>KLF4</i>-activated M2 macrophages accelerated the proliferation and migration of ESCs in vitro. <i>METTL14</i>-m6A-<i>KLF4</i> regulated macrophage polarization in EMs.</p></div>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":"197 8","pages":"5535 - 5551"},"PeriodicalIF":3.3,"publicationDate":"2025-06-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144473642","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":"Identification of Associations Between Peripheral Blood Gene Expression and Cerebrospinal Fluid Biomarkers in Alzheimer's Disease Using an Improved Joint Multi-Task Sparse Canonical Correlation Analysis Algorithm.","authors":"Qianqian Wu, Zhihui Ma, Feng Wang","doi":"10.1007/s12010-025-05297-y","DOIUrl":"https://doi.org/10.1007/s12010-025-05297-y","url":null,"abstract":"<p><p>Alzheimer's disease (AD) is an irreversible neurodegenerative disorder, and early diagnosis is crucial for effective clinical intervention. Traditional diagnostic methods involve detecting living brain tissue across the blood-brain barrier, but these invasive procedures cause unavoidable damage to patients. Genetic biomarkers in peripheral blood may provide valuable insights into brain lesions, potentially offering a non-invasive method for early AD diagnosis. The aim of this study is to propose an improved joint multi-task sparse canonical correlation analysis (MTSCCA) algorithm to identify significant genetic biomarkers in peripheral blood that correlate with brain markers of AD, such as cerebrospinal fluid (CSF) markers. This approach aims to accurately predict AD and assess disease progression. The study employs a multi-task sparse canonical correlation analysis (MTSCCA) approach with separate analyses for AD and healthy controls. Both tasks are constrained with class-consistent and class-specific conditions to identify significant features for each diagnostic group. To enhance robustness, the Laplacian matrix constraints were incorporated into the MTSCCA-LR algorithm to reduce noise in genetic data. The proposed algorithm identifies key differentially expressed genes (DEGs) that are involved in pathways closely linked to AD pathogenesis. These genes have specific diagnostic significance. Validation of these genes for predicting CSF markers was conducted using two regression models, showing good predictive accuracy. Furthermore, a Support Vector Machine (SVM) classifier was used to classify the two diagnostic groups, demonstrating high classification accuracy. The Top 20 genes identified using the proposed algorithm were used to construct an AD diagnostic model, which exhibited strong potential for non-invasive AD diagnosis, with significant implications for clinical practice. The code and example data of the proposed algorithm have been made publicly available on GitHub ( https://github.com/Zoe491/Improved-MTSCCA1 ).</p>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":" ","pages":""},"PeriodicalIF":3.1,"publicationDate":"2025-06-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144473630","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":"Combinational Therapy of Mesenchymal Stem Cells and Metformin in Bleomycin-Induced Idiopathic Pulmonary Fibrosis in Rat Model","authors":"Nourhan Hassan,&nbsp;Mariam N. Elbyoume,&nbsp;Mariam A. Taha,&nbsp;Hagar S. Mohamed,&nbsp;Omnia M. Elmoghini,&nbsp;Shorouk S. Raouf,&nbsp;Rwan K. Elsayem,&nbsp;Mohrail M. Medhat,&nbsp;Razan M. Rostom,&nbsp;Mohmed Hosney,&nbsp;Emad M. Elzayat","doi":"10.1007/s12010-025-05289-y","DOIUrl":"10.1007/s12010-025-05289-y","url":null,"abstract":"<div><p>Idiopathic pulmonary fibrosis (IPF) is a progressive and severe lung disease characterized by the buildup of interstitial fibrosis, where excessive collagen accumulates, leading to airway obstruction. This condition is initiated by the abnormal proliferation of alveolar type II (AT2) cells. Metformin, an established antidiabetic drug, has gained attention for its repurposed use as an anti-fibrotic agent. Meanwhile, adipose-derived mesenchymal stem cells (ADMSCs) exhibit potent anti-inflammatory and regenerative properties, and they have been shown to reduce collagen deposition. In this study, we hypothesize that the combination of metformin and ADMSCs can synergistically alleviate IPF and promote healthy lung tissue regeneration in a rat model. The goal is to evaluate the safety and efficacy of this approach at multiple levels; biochemical, molecular, histopathological, and histochemical. To induce IPF, Wistar albino rats received a single intratracheal dose of bleomycin (5 mg/kg body weight). The therapeutic phase involved treatment with either metformin or ADMSCs or a combination of both. Metformin was administered intraperitoneally (65 mg/kg body weight) every other day, while ADMSCs were delivered intravenously (1 × 10⁶ cells/0.5 ml DMEM/rat) through the tail vein. Our results demonstrated the effectiveness of combinational therapy, especially in mitigating oxidative stress. This was evidenced by the restoration of oxidative stress biomarkers, malondialdehyde (MDA), and catalase (CAT), as well as the regulation of collagenase type IV (MMP9), bovine serum albumin (BSA), and total protein levels in lung tissues. Moreover, the therapy modulated the expression of key inflammatory and fibrotic genes, including the pro-fibrotic marker TGF-β1, proinflammatory cytokine IL-6, and anti-inflammatory cytokine IL-10. Histopathological and histochemical analyses further supported the therapeutic benefits, showing significant recovery from bleomycin-induced fibrosis in rats treated with either the single or combined therapy. The findings suggest that this combinational approach could be a promising strategy for IPF treatment by simultaneously reducing inflammation, oxidative stress, and fibrosis while promoting tissue regeneration.</p><h3>Graphical Abstract</h3>\u0000<div><figure><div><div><picture><source><img></source></picture></div></div></figure></div></div>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":"197 8","pages":"5511 - 5534"},"PeriodicalIF":3.3,"publicationDate":"2025-06-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12334527/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144339697","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enhanced Nonaqueous Stability of Pseudomonas Cepacia Lipase Immobilized on Phthalocyanine-Dyed Absorbent Cotton","authors":"Yingchao Wang,&nbsp;Xing Zhao,&nbsp;Fangdi Cong,&nbsp;Wei Yang,&nbsp;Liyao Liu,&nbsp;Zhaowang Shen","doi":"10.1007/s12010-025-05295-0","DOIUrl":"10.1007/s12010-025-05295-0","url":null,"abstract":"<div><p>To enhance the nonaqueous catalysis of lipases, <i>Pseudomonas cepacia</i> lipase was immobilized on cotton fibers dyed with tetra-<i>β</i>-amino-phthalocyanine zinc or tetra-<i>β</i>-nitro-phthalocyanine zinc by physical adsorption in a column glass bottle and used to catalyze transesterification between hexanol and vinyl acetate. The suitable ratios of lipase to immobilized carriers were 10: 10 (mg: mg), and the employed carriers enhanced the enzymatic activity by more than 2.3-fold during the initial reaction stage. When the immobilized lipases were subjected to repeated uses at 37 °C and 160 rpm, their nonaqueous stability followed this order: cotton dyed with tetra-<i>β</i>-amino-phthalocyanine zinc &gt; cotton fibers &gt; cotton dyed with tetra-<i>β</i>-nitro-phthalocyanine zinc. The decrease in their substrate conversion efficiency after 24 h was 5.7%, 24.7%, and 33.3%, respectively, indicating that tetra-<i>β</i>-amino-phthalocyanine zinc provides effective protection for the enzyme. </p></div>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":"197 8","pages":"5494 - 5510"},"PeriodicalIF":3.3,"publicationDate":"2025-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144323993","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 Novel Esterase from Burkholderia sp. YD106 Capable of Hydrolysis of Methyl (R, S)-N-(2, 6-Dimethylphenyl) Alaninate, and Its Mutation for Improving Enantioselectivity","authors":"Ruixue Yang,&nbsp;Jianing Wu,&nbsp;Yunhe Zhang,&nbsp;Zhaohui Zhang","doi":"10.1007/s12010-025-05292-3","DOIUrl":"10.1007/s12010-025-05292-3","url":null,"abstract":"<div><p>Methyl (<i>R, S</i>)-2, 6-dimethylphenylaminopropionate ((<i>R, S</i>)-1) is an intermediate in the production of the agricultural fungicide (<i>R, S</i>)-metalaxyl. (<i>R, S</i>)-1 can be hydrolyzed enantioselectively by some hydrolases to produce (<i>R</i>)-1, which was used for the production of (<i>R</i>)-metalaxyl. In this work, a strain <i>Burkholderia</i> sp. YD106 that could hydrolyze (<i>R, S</i>)-1 was screened from the activated sludge, but it had almost no enantioselectivity. The intracellular active esterase WZest was successfully heterologously expressed in the recombinant <i>E. coli</i> BL21 (DE3)-pET-28a ( +)-GE04845. Using the recombinant strain as the parent strain, the mutants were constructed by a site-directed mutation. Among all 33 mutants, seven had altered enantioselectivity, of which four mutants were (<i>R</i>)-enantioselective and three were (<i>S</i>)-enantioselective. The mutant WZest-W23T had the highest (<i>R</i>)-enantioselectivity. When it catalyzed the hydrolysis of (<i>R, S</i>)-1 at 44.6% substrate conversion, <i>e.e.</i>_p reached 94.70% with an enantiomeric ratio (<i>E</i>) of 85.0. WZest showed significant amino acid sequence differences from the two reported esterases capable of hydrolyzing (<i>R, S</i>)-1. It was both active in two kinds of solutions. One was an emulsion with the substrate (<i>R, S</i>)-1 emulsified with Tween80, and the other was a homogeneous solution with acetone as a co-solvent. The activity of WZest in the former was higher than that in the latter.</p></div>","PeriodicalId":465,"journal":{"name":"Applied Biochemistry and Biotechnology","volume":"197 8","pages":"5452 - 5469"},"PeriodicalIF":3.3,"publicationDate":"2025-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144324091","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}