Process Biochemistry最新文献

An integrated study on the interactions of Lactobacillus brevis components with deoxynivalenol and zearalenone

IF 3.7 3区生物学

Process Biochemistry Pub Date : 2025-04-10 DOI: 10.1016/j.procbio.2025.04.005

Fatemeh Adami Ghamsari , Abdolamir Ghadaksaz , Maryam Tajabadi Ebrahimi , Maryam Bagheri Varzaneh , Alireza Iranbakhsh , Abbas Akhavan Sepahi

{"title":"An integrated study on the interactions of Lactobacillus brevis components with deoxynivalenol and zearalenone","authors":"Fatemeh Adami Ghamsari , Abdolamir Ghadaksaz , Maryam Tajabadi Ebrahimi , Maryam Bagheri Varzaneh , Alireza Iranbakhsh , Abbas Akhavan Sepahi","doi":"10.1016/j.procbio.2025.04.005","DOIUrl":"10.1016/j.procbio.2025.04.005","url":null,"abstract":"<div><div>Deoxynivalenol (DON) and zearalenone (ZEN) are toxic <em>Fusarium</em> mycotoxins that frequently contaminate food and feed, posing significant public health risks. Among mitigation strategies, biological methods using lactic acid bacteria (LAB; e.g., <em>Lactobacillus</em> spp.) show particular promise, as these microorganisms can effectively biodegrade mycotoxins into less toxic or inactive metabolites. This study explored the binding and biotransformation capabilities of <em>Lactobacillus brevis</em> components (S-layer, aryl-alcohol dehydrogenase, carboxylesterase, and lipoteichoic acid repeats) using computational docking and Fourier transform infrared (FTIR) spectroscopy. Docking simulations revealed stronger binding affinities for ZEN than DON, involving interactions with hydroxyl groups, oxygen atoms, alkyl chains, and aromatic rings. Enzymes formed stable complexes with mycotoxins, suggesting biotransformation potential. FTIR spectra after 24 hours of incubation revealed mycotoxin adsorption through interactions with bacterial cell wall components, indicated by C-H and C-C peak shifts. For DON, changes in OH and C<img>O peaks suggested oxidation-reduction, likely mediated by aryl-alcohol dehydrogenase, producing 3-keto-DON and 3-epi-DON. For ZEN, shifts in OH, C-O, and C<img>O peaks indicated lactone ring hydrolysis, likely catalyzed by carboxylesterase. These findings highlight <em>L. brevis</em> as a promising biocontrol agent that detoxifies DON and ZEN through binding and biotransformation, offering a potential strategy to mitigate mycotoxin contamination in food and feed.</div></div>","PeriodicalId":20811,"journal":{"name":"Process Biochemistry","volume":"154 ","pages":"Pages 12-21"},"PeriodicalIF":3.7,"publicationDate":"2025-04-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143823529","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Affinity resin selection for efficient capture of bispecific antibodies as guided by domain composition

IF 3.7 3区生物学

Process Biochemistry Pub Date : 2025-04-08 DOI: 10.1016/j.procbio.2025.04.007

Nattha Ingavat , Xinhui Wang , Yee Jiun Kok , Nuruljannah Dzulkiflie , Han Ping Loh , Eunice Leong , Kia Ngee Low , Amihan Anajao , Say Kong Ng , Yuansheng Yang , Xuezhi Bi , Wei Zhang

{"title":"Affinity resin selection for efficient capture of bispecific antibodies as guided by domain composition","authors":"Nattha Ingavat , Xinhui Wang , Yee Jiun Kok , Nuruljannah Dzulkiflie , Han Ping Loh , Eunice Leong , Kia Ngee Low , Amihan Anajao , Say Kong Ng , Yuansheng Yang , Xuezhi Bi , Wei Zhang","doi":"10.1016/j.procbio.2025.04.007","DOIUrl":"10.1016/j.procbio.2025.04.007","url":null,"abstract":"<div><div>Affinity chromatography is a cornerstone of bispecific antibody (bsAb) purification, with resin selection playing a critical role in developing downstream processes to ensure both process efficiency and product quality. Unlike monoclonal antibody purification, where Protein A chromatography is the gold standard for antibody capture, affinity chromatography in bsAb purification is often employed not only for capture but also for the removal of hard-to-eliminate product-related impurities. This study demonstrates that affinity resin selection can be effectively guided by analysing the domain composition of the target bsAb molecule and its potential impurities. Using faricimab, a CH1-CL CrossMab, as a model, Protein L — an affinity resin targeting the variable region of the light chain — was predicted to be the most effective affinity chromatography due to its different binding avidity towards faricimab from its major product-related impurities. Validation through screening four different types of affinity chromatography, each binding to distinct regions of faricimab, confirmed this prediction. Under optimized elution conditions, the purification process achieved ∼73 % purity from ∼30 % in the culture with ∼86 % monomeric yield, as well as decent removal of host cell proteins and host cell DNA.</div></div>","PeriodicalId":20811,"journal":{"name":"Process Biochemistry","volume":"154 ","pages":"Pages 1-11"},"PeriodicalIF":3.7,"publicationDate":"2025-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143821411","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Efficient removal of atrazine in wastewater by washed peanut shells biochar: Adsorption behavior and biodegradation

IF 3.7 3区生物学

Process Biochemistry Pub Date : 2025-04-08 DOI: 10.1016/j.procbio.2025.04.008

Yidan Wang , Yuhang Chen , Lianying Wang , Yayuan Mo , Xiangfeng Lin , Shu Gao , Menglin Chen

{"title":"Efficient removal of atrazine in wastewater by washed peanut shells biochar: Adsorption behavior and biodegradation","authors":"Yidan Wang , Yuhang Chen , Lianying Wang , Yayuan Mo , Xiangfeng Lin , Shu Gao , Menglin Chen","doi":"10.1016/j.procbio.2025.04.008","DOIUrl":"10.1016/j.procbio.2025.04.008","url":null,"abstract":"<div><div>Peanut shell powder was washed with water and then calcined to produce biochar, which acted as an adsorbent and biofilm supporter for the removal of atrazine (ATR). Biochar sample prepared by washing 6 h (named BC-W) achieved the highest biodegradation amount (<em>Q</em><sub><em>m</em></sub><em>,</em> 12.48 mg/g). The adsorption process of ATR on BC-W involved hydrogen bonding and π-π interactions. We found that the removal rate of ATR by biodegradation was 6.34–6.75 times that of adsorption, with an ATR concentration of 30 mg/L under half aeration. The results of high-throughput sequencing showed that there were two dominant bacterial phyla in the BC-W samples after biofilm culture: <em>Firmicutes</em> (51.89 %) and <em>Proteobacteria</em> (47.79 %), among which <em>Bacillus</em>, <em>Clostridium</em> (<em>sensu stricto</em>), and <em>Pseudomonas</em>, played a major role in the biodegradation of ATR. These findings showed that biochar could be employed as a low-cost adsorbent for the removal of pesticides from wastewater, also a biofilm supporter for enhanced biodegradation of organic pollutants.</div></div>","PeriodicalId":20811,"journal":{"name":"Process Biochemistry","volume":"154 ","pages":"Pages 22-34"},"PeriodicalIF":3.7,"publicationDate":"2025-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143825807","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Valorization of Solanum melongena L. crop by-products: Phenolic composition and in vitro antioxidant, antidiabetic, anti-inflammatory, cytotoxic, and antimicrobial properties

IF 3.7 3区生物学

Process Biochemistry Pub Date : 2025-04-07 DOI: 10.1016/j.procbio.2025.04.002

Mikel Añibarro-Ortega , Maria Inês Dias , Jovana Petrović , Sonia Núñez , Ricardo C. Calhelha , Eduardo M. Costa , M. Machado , Manuela Pintado , Marina Soković , Víctor López , Lillian Barros , José Pinela

{"title":"Valorization of Solanum melongena L. crop by-products: Phenolic composition and in vitro antioxidant, antidiabetic, anti-inflammatory, cytotoxic, and antimicrobial properties","authors":"Mikel Añibarro-Ortega , Maria Inês Dias , Jovana Petrović , Sonia Núñez , Ricardo C. Calhelha , Eduardo M. Costa , M. Machado , Manuela Pintado , Marina Soković , Víctor López , Lillian Barros , José Pinela","doi":"10.1016/j.procbio.2025.04.002","DOIUrl":"10.1016/j.procbio.2025.04.002","url":null,"abstract":"<div><div>This study explored the valorization of post-harvest eggplant aerial parts as a sustainable source of value-added ingredients by investigating their phenolic composition and <em>in vitro</em> bioactive properties. HPLC-DAD-ESI/MS<sup><em>n</em></sup> analysis identified chlorogenic acid derivatives as the predominant phenolic compounds (53 % of the phenolic fraction), followed by <em>O</em>-glycosylated kaempferol and quercetin. The extract displayed antioxidant activity in physiologically relevant cell-based assays and significant α-glucosidase inhibitory capacity that far exceeded that of the standard drug acarbose. It also inhibited the formation of advanced glycation end-products (AGEs), suggesting its potential to mitigate diabetes-related complications. Furthermore, the extract showed a modest pancreatic lipase inhibitory effect and capacity to suppress interleukin 6 production. Selective cytotoxicity against human gastric and colon adenocarcinoma cell lines and strong antimicrobial activity against foodborne pathogens were observed. Given the growing demand for natural alternatives to synthetic drugs, these findings position eggplant crop biomass as a promising, sustainable source of active compounds with potential applications in food, nutraceutical, and pharmaceutical formulations for managing type 2 diabetes and other oxidative stress-mediated conditions. This study not only contributes to the valorization of agricultural waste but also expands the research on by-products of Solanaceae crops, offering a pathway for sustainable resource utilization.</div></div>","PeriodicalId":20811,"journal":{"name":"Process Biochemistry","volume":"153 ","pages":"Pages 315-324"},"PeriodicalIF":3.7,"publicationDate":"2025-04-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143816455","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Effect of lactic acid bacteria fermentation on biotransformation of phenolic compounds and bioactivities of hawthorn pulp based on metabolomics

IF 3.7 3区生物学

Process Biochemistry Pub Date : 2025-04-04 DOI: 10.1016/j.procbio.2025.03.021

Wanting Zhang, Xiaomin Yang, Yan Xu, Bingjie Liu, Xianghong Meng

{"title":"Effect of lactic acid bacteria fermentation on biotransformation of phenolic compounds and bioactivities of hawthorn pulp based on metabolomics","authors":"Wanting Zhang, Xiaomin Yang, Yan Xu, Bingjie Liu, Xianghong Meng","doi":"10.1016/j.procbio.2025.03.021","DOIUrl":"10.1016/j.procbio.2025.03.021","url":null,"abstract":"<div><div>Lactic acid bacteria (LAB) fermentation can offer an improvement in the nutritional quality and biological activity of foods. This study aimed to explain the impacts of LAB fermentation on bioactivities and biotransformation of polyphenols in hawthorn pulp. Firstly, a strain with better polyphenol transformation capacity was selected from four LAB strains; then, microbial characteristics, polyphenols content, antioxidant capacity, and digestive enzymes inhibitory capacity of fermented hawthorn pulp were evaluated and their correlation was established; finally, metabolomics was used for analyzing the biotransformation of polyphenols. The results showed that <em>Lactobacillus rhamnosus</em> FN518 exhibited the best efficiency, which reached viable counts of 8.28 ± 0.03 log CFU/mL after 48 h of fermentation and exhibited strong malolactic-conversion ability. The total phenolic content significantly increased to 11.25 ± 0.12 mg GAE/g and DPPH radical scavenging capacity, FRAP, and α-amylase inhibitory capacity significantly increased by 30.80 %, 13.21 %, and 16.82 %, respectively (<em>p</em> < 0.05), which were significantly and positively correlated with phenolic compounds. Metabolomics revealed that flavonoid biosynthesis and enzymatic degradation of macromolecular phenolic compounds were primary biotransformation pathways of polyphenols in fermented hawthorn pulp. This study contributes precious insights into biotransformation of hawthorn polyphenols via LAB fermentation and development of fermented hawthorn products.</div></div>","PeriodicalId":20811,"journal":{"name":"Process Biochemistry","volume":"153 ","pages":"Pages 304-314"},"PeriodicalIF":3.7,"publicationDate":"2025-04-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143808155","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

A mechanistic insight into polyethylene degradation by Bacillus sp. and, Lysinibacillus sp. from mangrove soil

IF 3.7 3区生物学

Process Biochemistry Pub Date : 2025-04-04 DOI: 10.1016/j.procbio.2025.03.018

N.D. Dhanraj , U.P. Sreelakshmi , P. Sneha , M.S. Jisha

{"title":"A mechanistic insight into polyethylene degradation by Bacillus sp. and, Lysinibacillus sp. from mangrove soil","authors":"N.D. Dhanraj , U.P. Sreelakshmi , P. Sneha , M.S. Jisha","doi":"10.1016/j.procbio.2025.03.018","DOIUrl":"10.1016/j.procbio.2025.03.018","url":null,"abstract":"<div><div>This study aims to establish a sustainable method for plastic remediation, particularly focusing on polyethylene (PE). Bacteria capable of breaking down PE were identified and characterized from plastic-contaminated mangrove soils in Mangalavanam and Puthuvype, where limited efforts had been made to explore PE-degrading microbes. Two isolates, Ma21 and Pu9, showed efficient PE degradation, which exhibited 7.8 % and 4.2 % weight loss of PE film respectively. This degradation was supported by FTIR analysis, indicating initial degradation steps, and FE-SEM analysis, showing surface deterioration of PE films due to bacterial action. GC-MS analysis of reaction intermediates detected phthalic acid compounds, suggesting the ability of these bacteria to degrade PE and indicating the presence of plastic additives in the degradation medium. Screening for biosurfactant production revealed that both Ma21 and Pu9 can produce biosurfactants. The production of alkane hydroxylase enzyme and laccase enzyme by Ma21 and Pu9 further supported their biodegradation capabilities. Molecular identification through 16S rDNA sequencing confirmed that Ma21 is <em>Bacillus thuringiensis</em>, and Pu9 is <em>Lysinibacillus boronitolerans</em>.</div></div>","PeriodicalId":20811,"journal":{"name":"Process Biochemistry","volume":"153 ","pages":"Pages 294-303"},"PeriodicalIF":3.7,"publicationDate":"2025-04-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143808156","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Enhanced 2-phenylethanol and co-production of pullulan from synthetic medium by Aureobasidium pullulans

IF 3.7 3区生物学

Process Biochemistry Pub Date : 2025-04-04 DOI: 10.1016/j.procbio.2025.03.025

Lijun Wu , Guoqiang Chen , Xinruitong Liu , Yuxi Wei , Lijuan Liu , Weiyao Hu , Haibo Zhang

{"title":"Enhanced 2-phenylethanol and co-production of pullulan from synthetic medium by Aureobasidium pullulans","authors":"Lijun Wu , Guoqiang Chen , Xinruitong Liu , Yuxi Wei , Lijuan Liu , Weiyao Hu , Haibo Zhang","doi":"10.1016/j.procbio.2025.03.025","DOIUrl":"10.1016/j.procbio.2025.03.025","url":null,"abstract":"<div><div>2-phenylethanol (2-PE) and its derivatives play crucial roles in food flavoring and fine chemical synthesis. However, their industrial-scale production yields remain a key constraint for broader industrial implementation. To overcome this challenge, we implemented an integrated <em>in-situ</em> product adsorption (ISPR) strategy combined with nitrogen source restriction and a high-glucose medium formulation during 2-PE biosynthesis using <em>Aureobasidium pullulans</em> (<em>A</em>. <em>pullulans</em>) A5. Notably, this approach enabled simultaneous co-production of 2-PE and the commercially valuable exopolysaccharide pullulan. The experimental protocol comprised two phases: First, we established a basal production medium (medium-1) containing 20 % (w/v) glucose and 1.8 % (w/v) Yeast Nitrogen Base (YNB). Subsequently, we systematically screened adsorption agents, identifying Trialkylphosphine Oxide (TRPO) as an optimal extractant that demonstrated selective 2-PE adsorption capacity but not for L-phenylalanine (L-phe). In the optimized system, supplementation of 10 mL TRPO to 50 mL medium-1 (pH 6.0) supplemented with 4 g/L of L-phe as precursor resulted in 2.39 g/L 2-PE production (60 % molar conversion efficiency) and 5.16 g/L pullulan accumulation after 48 h fermentation. Process analysis revealed that TRPO-mediated ISPR enhanced 2-PE volumetric productivity, while the high-carbon medium configuration synergistically increased both target metabolite titers. These technical improvements significantly improved process economics while simultaneously streamlining downstream processing for industrial-scale operations</div></div>","PeriodicalId":20811,"journal":{"name":"Process Biochemistry","volume":"153 ","pages":"Pages 259-266"},"PeriodicalIF":3.7,"publicationDate":"2025-04-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143776735","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Phosphorus and sulphur affecting the growth, nutrient uptake and amino acids composition in Dunaliella salina

IF 3.7 3区生物学

Process Biochemistry Pub Date : 2025-04-02 DOI: 10.1016/j.procbio.2025.03.026

Yixing Sui , Gleison de Souza Celente , Laurens Bosscher Navarro , Pablo Garcia-Trinanes , Patricia J. Harvey

{"title":"Phosphorus and sulphur affecting the growth, nutrient uptake and amino acids composition in Dunaliella salina","authors":"Yixing Sui , Gleison de Souza Celente , Laurens Bosscher Navarro , Pablo Garcia-Trinanes , Patricia J. Harvey","doi":"10.1016/j.procbio.2025.03.026","DOIUrl":"10.1016/j.procbio.2025.03.026","url":null,"abstract":"<div><div>The potential of microalga <em>Dunaliella salina</em> as a protein source and in bioremediation application is still underdeveloped, despite its known carotenoids overproduction. This study focused on investigating the effects of phosphorus (P) and sulphur (S) concentrations on the biomass growth and essential amino acids profile of <em>D. salina</em> PLY DF15 (CCAP 19/41). Additionally, its nutrient removal capacity was analysed in a bioremediation context. From all tested P and S concentrations, <em>D. salina</em> resulted in high-quality protein with essential amino acid index (EAAI) between 1.6 and 1.7 (EAAI ≥ 1 meets FAO/WHO standard) with little variations from all P and S concentrations. In a bioremediation context, <em>D. salina</em> also showed great potential for nutrients removal and recovery with high adaptability to varying nutrient conditions, with overall more than 80 % and 100 % nitrogen (N) and P recovery, respectively. The full recovery of S however was only achieved at lower S levels. Overall <em>D. salina</em> shows great potential in providing high-quality protein for human consumption using economic and sustainable cultivation approach.</div></div>","PeriodicalId":20811,"journal":{"name":"Process Biochemistry","volume":"153 ","pages":"Pages 276-283"},"PeriodicalIF":3.7,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143791203","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

One-pot chemo-enzymatic cascades using ene-reductase and alcohol dehydrogenase to produce a brivaracetam chiral precursor

IF 3.7 3区生物学

Process Biochemistry Pub Date : 2025-04-02 DOI: 10.1016/j.procbio.2025.04.001

Jiacheng Feng , Dingjun Chu , Ruiwei Chen , Xiaoqiang Xie , Yongxian Fan , Xiaolong Chen , Linjiang Zhu

{"title":"One-pot chemo-enzymatic cascades using ene-reductase and alcohol dehydrogenase to produce a brivaracetam chiral precursor","authors":"Jiacheng Feng , Dingjun Chu , Ruiwei Chen , Xiaoqiang Xie , Yongxian Fan , Xiaolong Chen , Linjiang Zhu","doi":"10.1016/j.procbio.2025.04.001","DOIUrl":"10.1016/j.procbio.2025.04.001","url":null,"abstract":"<div><div>Brivaracetam is a third-generation antiepileptic drug containing two chiral centers (<em>2S</em>, <em>4 R</em>). To achieve effective asymmetric synthesis of 4 <em>R</em> chiral center, a novel one-pot chemo-enzymatic cascade strategy was developed to synthesize the 4 <em>R</em> chiral precursor, (<em>R</em>)-4-propyldihydrofuran-2(3<em>H</em>)-one (<strong>1b</strong>). The butenolide 5-hydroxy-4-propylfuran-2(5<em>H</em>)-one was selected as the starting substrate due to the presence of an active electron-withdrawing group, which enabled rapid enzymatic reduction by an ene-reductase with > 99 % conversion. However,‌ this reaction generated ‌a racemization-prone intermediate‌. Subsequent reduction ‌using a newly identified alcohol dehydrogenase (YahK)‌ demonstrated high activity but ‌exhibited <em>S</em>-configuration stereoselectivity‌. ‌ Through semi-rational engineering, the stereoselectivity of a YahK triple mutant (G132T/T182A/M313R) was successfully reversed to favor the <em>R</em>-configuration. The chemo-enzymatic cascade combining ADH reduction and chemical cyclization yielded the chiral product <em>4R</em>-<strong>1b</strong> with an enantiomeric excess (<em>ee</em>) of > 91 % (<em>R</em>). Notably, the one-pot mode further enhanced the <em>ee</em> value of the final product to > 98 %. This strategy effectively bypassed the limitations of low-reactivity alkene substrates, enabling efficient construction of the <em>4 R</em> chiral center in brivaracetam.</div></div>","PeriodicalId":20811,"journal":{"name":"Process Biochemistry","volume":"153 ","pages":"Pages 267-275"},"PeriodicalIF":3.7,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143785098","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Shedding light on H. pylori detection: A fusion protein approach unveiled through LIPS method

IF 3.7 3区生物学

Process Biochemistry Pub Date : 2025-04-01 DOI: 10.1016/j.procbio.2025.03.023

Seyedeh Mahsa Farzanfar , Sedigheh Asad

{"title":"Shedding light on H. pylori detection: A fusion protein approach unveiled through LIPS method","authors":"Seyedeh Mahsa Farzanfar , Sedigheh Asad","doi":"10.1016/j.procbio.2025.03.023","DOIUrl":"10.1016/j.procbio.2025.03.023","url":null,"abstract":"<div><div>The Luciferase Immunoprecipitation Systems (LIPS) method is a highly sensitive approach for quantitatively detecting antibodies, offering potential in identifying viral and bacterial infections. However, the substantial size of the luciferase-antigen fusion protein presents challenges in production and folding. Using epitopes rather than the full-length antigenic protein may circumvent issues with recombinant expression. <em>Helicobacter pylori</em>, a gram-negative bacterium, poses a risk of gastric cancer if untreated. This study focuses on producing a fusion protein comprising in silico-designed antigenic epitopes from the <em>H. pylori</em> urease protein and luciferase, aiming to reduce the fusion protein's size and augment its expression in the <em>E. coli</em> system. Bioinformatic analysis identified sequences encoding antigenic regions, which were amplified via PCR. A luciferase-linker-epitope construct was then devised and expressed in the <em>E. coli Bl21</em> (DE3) strain. The recombinant protein was primarily purified to homogeneity, yielding a major band at 75 kilodaltons. Verification of the protein's proper folding and functionality was confirmed through a bioluminescence assay with an emission of 13.7 × 10^6 RLU/s. Western blot analysis authenticated the fusion protein's specific binding to <em>H. pylori</em> antibodies. These findings underscore the potential of the protein as a promising candidate for <em>H. pylori</em> detection and streamline LIPS fusion protein production.</div></div>","PeriodicalId":20811,"journal":{"name":"Process Biochemistry","volume":"153 ","pages":"Pages 212-219"},"PeriodicalIF":3.7,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143768998","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0