Journal of biotechnology最新文献_第3页

Gene hunting and semi-rational design of β-xylosidase from Aspergillus aculeatus for highly efficient hydrolysis of astragaloside 刺曲霉β-木糖苷酶高效水解黄芪甲苷的基因寻找与半合理设计。

IF 3.9 2区生物学

Journal of biotechnology Pub Date : 2025-09-06 DOI: 10.1016/j.jbiotec.2025.09.001

Yuanyuan Zheng, Huaxia Cao, Yu Liu, Feng Xue

{"title":"Gene hunting and semi-rational design of β-xylosidase from Aspergillus aculeatus for highly efficient hydrolysis of astragaloside","authors":"Yuanyuan Zheng, Huaxia Cao, Yu Liu, Feng Xue","doi":"10.1016/j.jbiotec.2025.09.001","DOIUrl":"10.1016/j.jbiotec.2025.09.001","url":null,"abstract":"<div><div>Cycloastragenol (CA), the triterpenoid aglycone of astragaloside (ASI), is a telomerase activator and potential anti-aging drug with broad application prospects. Due to the rapid increase of its market demand in recent years, efficient production of CA has attracted increasing attention. In this study, the novel β-xylosidase XylO2 from <em>Aspergillus aculeatus</em> was identified through genome mining. It is capable of hydrolyzing ASI into cycloastragenol-6-O-β-D-glucoside (CMG), a significant precursor in the preparation of CA. Subsequently, structure-guided semi-rational design and virtual screening were used to obtain the highly active variant C23T, which exhibited more than 1.94 and 2.10-fold improvement of catalytic activity with ASI and <em>p</em>-nitrophenyl-β-D-xylopyranoside, respectively. According to structural modelling and molecule docking, the reduced volume of the binding pocket as well as the decreased distance between the substrate and active site residues positively influenced the catalytic activity. Moreover, the conversion yield of 1 g/L ASI increased to 99.10 %, suggesting the potential application of this variant in CA production. This study not only provides a novel enzyme for glycoside hydrolysis, but also offers theoretical insights for understanding and modifying these enzymes.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"408 ","pages":"Pages 80-90"},"PeriodicalIF":3.9,"publicationDate":"2025-09-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145023402","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Optimizing lignocellulose conversion: A comparative study of alkali-assisted ball milling pretreatment of cotton stalk and corn stover 优化木质纤维素转化：碱辅助球磨预处理棉秆和玉米秸秆的比较研究

IF 3.9 2区生物学

Journal of biotechnology Pub Date : 2025-09-04 DOI: 10.1016/j.jbiotec.2025.08.018

Hui Zhang , Xiwen Jia , Kaili Ding , Xueyan Liang , Yinghui He , Xiaojun Guo , Ling Zhou , Lujia Han , Weihua Xiao

{"title":"Optimizing lignocellulose conversion: A comparative study of alkali-assisted ball milling pretreatment of cotton stalk and corn stover","authors":"Hui Zhang , Xiwen Jia , Kaili Ding , Xueyan Liang , Yinghui He , Xiaojun Guo , Ling Zhou , Lujia Han , Weihua Xiao","doi":"10.1016/j.jbiotec.2025.08.018","DOIUrl":"10.1016/j.jbiotec.2025.08.018","url":null,"abstract":"<div><div>Cotton stalk (CTS) and corn stover (CRS) were pretreated using solid alkali (NaOH or Ca(OH)<sub>2</sub>) assisted ball milling (BM). The physicochemical properties of the pretreated materials and their high-solid enzymatic hydrolysis performance were systematically investigated. The interaction between alkali and straw was synergistically enhanced by mechanical force generated during BM, achieving effective lignin removal. NaOH showed superior delignification efficiency compared to Ca(OH)<sub>2</sub> for both straws at same dosages. Analysis of cation exchange capacity revealed higher stability of CTS-Ca<sup>2</sup><sup>+</sup> complex than CRS-Ca<sup>2+</sup> complex, explaining the more effective lignin removal from CTS than from CRS by Ca(OH)<sub>2</sub>. During the alkali/BM process, cellulose showed structural swelling and crystalline destruction. The cellulose of CTS mainly retained its cellulose I crystallinity, whereas most of the cellulose of CRS was converted to an amorphous state. The changes in physicochemical properties and structure effectively improved the enzymatic saccharification efficiency of pretreated straws. Under high-solids loading conditions (20 %) with an enzyme loading of 20 FPU/g, the maximum glucose yields from NaOH-pretreated and Ca(OH)<sub>2</sub>-pretreated CTS were 68.52 % and 65.90 %, respectively. The corresponding yields for CRS were 80.43 % and 80.30 %. Partial least squares (PLS) analysis identified straw composition, particularly changes in cellulose, as the dominant factor influencing enzymatic hydrolysis efficiency. This study provided valuable insights for optimizing biomass-specific pretreatment strategies to achieve efficient sugar production.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"408 ","pages":"Pages 15-27"},"PeriodicalIF":3.9,"publicationDate":"2025-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144997605","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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 novel dynamic flux balance analysis for modeling CHO cell fed-batch cultures with pH and temperature shifts 一种新的动态通量平衡分析，用于模拟pH和温度变化的CHO细胞补料批培养。

IF 3.9 2区生物学

Journal of biotechnology Pub Date : 2025-09-04 DOI: 10.1016/j.jbiotec.2025.08.010

Ali Ghodba , Anne Richelle , Chris McCready , Luis Ricardez-Sandoval , Hector Budman

{"title":"A novel dynamic flux balance analysis for modeling CHO cell fed-batch cultures with pH and temperature shifts","authors":"Ali Ghodba , Anne Richelle , Chris McCready , Luis Ricardez-Sandoval , Hector Budman","doi":"10.1016/j.jbiotec.2025.08.010","DOIUrl":"10.1016/j.jbiotec.2025.08.010","url":null,"abstract":"<div><div>While Dynamic Flux Balance Analysis provides a powerful framework for simulating metabolic behavior, incorporating operating conditions such as pH and temperature, which profoundly impact monoclonal antibodies production, remains challenging. This study presents an advanced dFBA model that integrates kinetic constraints formulated as functions of pH and temperature to predict CHO cell metabolism under varying operational conditions. The model was validated against data from 20 fed-batch experiments conducted in Ambr®250 bioreactors. To mitigate overparameterization, a bi-level optimization approach utilizing the Bayesian Information Criterion was employed to systematically identify the most effective kinetic constraints. This optimization reduced the number of parameters (from 253 to 205) while improving predictive accuracy by up to 8.3% for training and 2.68% for validation datasets. The results highlight the model’s ability to predict cell growth, titer, and also capture metabolic shifts, including glucose, lactate, and ammonia metabolism and amino acid utilization, across different temperature and pH conditions with high predictive precision (average <span><math><mrow><msup><mrow><mi>R</mi></mrow><mrow><mn>2</mn></mrow></msup><mo>≥</mo><mn>0</mn><mo>.</mo><mn>97</mn></mrow></math></span> for cell growth and titer and average <span><math><mrow><msup><mrow><mi>R</mi></mrow><mrow><mn>2</mn></mrow></msup><mo>≥</mo><mn>0</mn><mo>.</mo><mn>85</mn></mrow></math></span> for other metabolites). This optimized dFBA framework offers a robust tool for studying model-based optimization for CHO cell metabolism, identifying optimal operating conditions to balance growth and productivity.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"408 ","pages":"Pages 61-71"},"PeriodicalIF":3.9,"publicationDate":"2025-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145008251","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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 review on microbial metabolic engineering for the improvement of itaconic acid production 改善衣康酸生产的微生物代谢工程研究进展

IF 3.9 2区生物学

Journal of biotechnology Pub Date : 2025-09-03 DOI: 10.1016/j.jbiotec.2025.08.017

Yanhua Zhang , Tianqi Han , Wenlong Qu , Xiaolu Wang , Depei Wang , Xianli Xue

{"title":"A review on microbial metabolic engineering for the improvement of itaconic acid production","authors":"Yanhua Zhang , Tianqi Han , Wenlong Qu , Xiaolu Wang , Depei Wang , Xianli Xue","doi":"10.1016/j.jbiotec.2025.08.017","DOIUrl":"10.1016/j.jbiotec.2025.08.017","url":null,"abstract":"<div><div>Itaconic acid (IA), also known as methylene-butanedioic acid, is one of the top 12 platform chemicals in the world and demand for it is growing worldwide. The itaconic acid molecule contains one reactive double bond and two carboxyl groups, endowing it with chemical reactivity. It is widely used in fields such as light industry, agriculture, energy, and medical applications. The recent progress in understanding the biosynthesis, the regulation and the cellular transport of itaconic acid has facilitated the optimisation of existing processes as well as the construction of new microbial platforms. However, there is still need for further optimisation to increase the space-time yield, to achieve higher final concentrations and to use a broader range of low cost sustainable raw materials. In this review article, we have summarized the current status of itaconic acid production by natural microbial strains, recent advances in research on genetically engineered modified strains, and further analyzed the gene clusters associated with itaconic acid biosynthesis in <em>Aspergillus terreus</em>, <em>Ustilago maydis</em>, as well as similar genetic clusters identified in <em>A. niger</em>. Genetic engineering and process development need to apply intelligent screening platforms to obtain as much information as possible in small scale so as to provide more detailed basic information and newer improvement ideas for subsequent research.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"408 ","pages":"Pages 28-38"},"PeriodicalIF":3.9,"publicationDate":"2025-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145004677","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Preparation of high-purity 1,3-distearoyl-2-oleoylglycerol (StOSt) via a two-step enzymatic interesterification combined with two-step solvent fractionation 两步酶催化酯化结合两步溶剂分离制备高纯度1,3-二硬脂酰-2-油基甘油（StOSt）

IF 3.9 2区生物学

Journal of biotechnology Pub Date : 2025-09-03 DOI: 10.1016/j.jbiotec.2025.08.016

Chaeyeon Lee , Sopark Sonwai , Junsoo Lee , Byung Hee Kim , Heon-Woong Kim , In-Hwan Kim

{"title":"Preparation of high-purity 1,3-distearoyl-2-oleoylglycerol (StOSt) via a two-step enzymatic interesterification combined with two-step solvent fractionation","authors":"Chaeyeon Lee , Sopark Sonwai , Junsoo Lee , Byung Hee Kim , Heon-Woong Kim , In-Hwan Kim","doi":"10.1016/j.jbiotec.2025.08.016","DOIUrl":"10.1016/j.jbiotec.2025.08.016","url":null,"abstract":"<div><div>High-purity 1,3-distearoyl-2-oleoylglycerol (StOSt) was produced from high-oleic sunflower oil and ethyl stearate via a two-step enzymatic interesterification combined with a two-step solvent fractionation. Lipozyme RM IM (Rhizomucor miehei) was employed as a biocatalyst and the reaction was conducted in a packed-bed reactor. Molecular distillation was used to remove fatty acid ethyl esters from the reaction mixtures after enzymatic reactions. The optimum conditions were a temperature of 55 °C, a water content of 300 ppm (based on the total weight of the substrate), and a molar ratio of 1:8 (triacylglycerol (TAG) to ethyl stearate). The second enzymatic interesterification was carried out under the optimum conditions, resulting in the synthesis of 67.8 % StOSt-enriched TAG. To further enrich StOSt, two rounds of solvent fractionation were conducted using n-hexane and acetone, respectively. Consequently, StOSt TAG with a high purity of 83 % was achieved through the processes designed in this study.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"408 ","pages":"Pages 50-60"},"PeriodicalIF":3.9,"publicationDate":"2025-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145005135","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Enhancement and mechanism of in-cluster two-component regulatory factors on granaticin production in Streptomyces vilmorinianum YP1 簇内双组分调控因子对vilmorinianum Streptomyces YP1 Granaticin生产的增强作用及其机制

IF 3.9 2区生物学

Journal of biotechnology Pub Date : 2025-08-30 DOI: 10.1016/j.jbiotec.2025.08.015

Bingyu Zhao , Qiaolei Zhu , Xin Wang , Xiaoya Shang , Yunping Zhu

{"title":"Enhancement and mechanism of in-cluster two-component regulatory factors on granaticin production in Streptomyces vilmorinianum YP1","authors":"Bingyu Zhao , Qiaolei Zhu , Xin Wang , Xiaoya Shang , Yunping Zhu","doi":"10.1016/j.jbiotec.2025.08.015","DOIUrl":"10.1016/j.jbiotec.2025.08.015","url":null,"abstract":"<div><div>Granaticins, a class of bioactive benzoisochromanequinones (BIQs), are natural blue pigments with biological activities, including antibacterial properties and promising clinical anticancer applications. However, their clinical and industrial applications are limited by low production yields and unclear biosynthetic regulation. In this study, we identified a rare in-cluster two-component system (TCS; <em>orf10</em>/<em>orf11</em>) within the granaticin biosynthesis-related gene cluster (BGC) of <em>Streptomyces vilmorinianum</em> YP1, a novel high-yielding producer. The <em>orf10</em> encodes a MerR family transcription regulator, while the <em>orf11</em> encodes a sensor histidine kinase. The roles of <em>orf10</em> and <em>orf11</em> were determined using single and double knockouts and complementation experiments. The yield of granaticins increased with the overexpression of the two genes in <em>S. vilmorinianum</em> YP1. Notably, <em>orf10</em> overexpression resulted in a record high granaticin B titer of 716.27 mg/L, representing a substantial improvement over native production level. The metabolic map obtained using Ultra Performance Liquid Chromatography Tandem Mass Spectrometry (UPLC-MS/MS) analysis revealed that the metabolic processes of the combined overexpression mutant were significantly different from those of the single overexpression mutants. These findings not only establish the functional link between the in-cluster TCSs, and granaticin biosynthesis but also provide an effective engineering strategy to overcome production bottlenecks. Our work advances the mechanistic understanding of BIQ regulation and highlights <em>S. vilmorinianum</em> YP1 as a promising platform for industrial-scale granaticin production.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"408 ","pages":"Pages 39-49"},"PeriodicalIF":3.9,"publicationDate":"2025-08-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144955211","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Metabolic engineering of Vibrio natriegens for the production of N-acetylglucosamine 产n -乙酰氨基葡萄糖弧菌的代谢工程

IF 3.9 2区生物学

Journal of biotechnology Pub Date : 2025-08-29 DOI: 10.1016/j.jbiotec.2025.08.012

Yanjie Li , Jiujiu Yi , Zheng-Jun Li

{"title":"Metabolic engineering of Vibrio natriegens for the production of N-acetylglucosamine","authors":"Yanjie Li , Jiujiu Yi , Zheng-Jun Li","doi":"10.1016/j.jbiotec.2025.08.012","DOIUrl":"10.1016/j.jbiotec.2025.08.012","url":null,"abstract":"<div><div><em>Vibrio natriegens</em> has emerged as a promising microbial chassis for industrial biotechnology due to its unparalleled growth rate and metabolic versatility. In this study, <em>V. natriegens</em> ATCC14048 was rationally engineered as a high-efficiency platform for <em>N</em>-acetylglucosamine (GlcNAc) biosynthesis. First, key biosynthetic genes including <em>glmS</em> from <em>Escherichia coli</em> and <em>Scgna1</em> from <em>Saccharomyces cerevisiae</em> were overexpressed, while the native GlcNAc degradation gene <em>nagA</em> was knocked out, which led to a GlcNAc titer of 0.11 g/L. Subsequently, blocking the pentose phosphate pathway and glycolysis enhanced precursor availability and achieved GlcNAc titer of 1.22 g/L. Metabolic flux was further amplified by deleting <em>rapZ</em>, <em>nagB</em>, and <em>manX</em> genes, along with chromosomal integration of the strong promoter J23119 upstream of endogenous <em>glmS</em>, yielding 3.59 g/L GlcNAc. Shake flask cultivation optimization through nitrogen source addition and oxygen supply ultimately elevated GlcNAc titer to 6.89 g/L in shake-flask cultures. This work demonstrates <em>V. natriegens</em>' exceptional metabolic plasticity and positions it as a superior chassis for industrial-scale biomanufacturing of amino sugars.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"407 ","pages":"Pages 105-110"},"PeriodicalIF":3.9,"publicationDate":"2025-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144922568","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

3D cell spheroid inoculated with bacteria: An in vitro model for assessing antimicrobial efficacy 接种细菌的三维细胞球体：一种评估抗菌效果的体外模型

IF 3.9 2区生物学

Journal of biotechnology Pub Date : 2025-08-29 DOI: 10.1016/j.jbiotec.2025.08.008

Xiaoyan Yang, Ke Ning, Lingke Feng, Yirong Chen, Yuanyuan Xie, Mingzhu Wang, Ling Yu

{"title":"3D cell spheroid inoculated with bacteria: An in vitro model for assessing antimicrobial efficacy","authors":"Xiaoyan Yang, Ke Ning, Lingke Feng, Yirong Chen, Yuanyuan Xie, Mingzhu Wang, Ling Yu","doi":"10.1016/j.jbiotec.2025.08.008","DOIUrl":"10.1016/j.jbiotec.2025.08.008","url":null,"abstract":"<div><div>Bacterial infections persist as a significant global health challenge, intensifying the demand for novel antimicrobial agents capable of overcoming persistent infections and mitigating the spread of drug-resistant strains. Traditional 2D cell culture assays, prone to bacterial contamination, fail to recapitulate the complex 3D architecture of <em>in vivo</em> tissues, rendering them inadequate as <em>in vitro</em> models for evaluating antimicrobial efficacy. This study investigates the effectiveness of 3D spheroids inoculated with bacteria, hypothesizing that 3D spheroids allow for assessment of antibacterial agents. Human prostate cancer cells (DU145) were cultured into 7-day-old spheroids. The morphology and migration ability of the 3D spheroids before and after inoculation with the model bacterium <em>E. coli</em> were analyzed. Through fluorescence microscopy and single-cell multimode analysis, it was found that <em>E. coli</em> can penetrate into the central region of the 3D spheroid while maintaining its viability. The complex multicellular structure of the 3D spheroids is retained under bacterial growth pressure, and the migration function of the 3D spheroids is preserved. The antimicrobial efficacy of the commercial antibiotic gentamicin and the functional material ZIF-8@Ag was evaluated by counting viable <em>E. coli</em> within the 3D spheroids. The results showed that both gentamicin and ZIF-8@Ag can penetrate the 3D spheroids, eradicating the embedded <em>E. coli</em> while preserving the spheroid’s structure. These findings highlight the importance of 3D spheroid-bacteria co-culture models as valuable tools for evaluating the permeability and efficacy of antimicrobial agents.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"407 ","pages":"Pages 97-104"},"PeriodicalIF":3.9,"publicationDate":"2025-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144922569","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Microalgae in 3D bioprinting and bone regeneration: Trends, applications, and future perspectives 微藻在3D生物打印和骨再生：趋势，应用和未来展望。

IF 3.9 2区生物学

Journal of biotechnology Pub Date : 2025-08-29 DOI: 10.1016/j.jbiotec.2025.08.011

Qi Zhang , Gang Wang , Yuelei Zhang , Sheng Huang , Jiangxin Wang

{"title":"Microalgae in 3D bioprinting and bone regeneration: Trends, applications, and future perspectives","authors":"Qi Zhang , Gang Wang , Yuelei Zhang , Sheng Huang , Jiangxin Wang","doi":"10.1016/j.jbiotec.2025.08.011","DOIUrl":"10.1016/j.jbiotec.2025.08.011","url":null,"abstract":"<div><div>3D bioprinting has emerged as a powerful technology for bone tissue engineering, enabling the fabrication of complex, cell-laden structures with spatial precision. In recent years, microalgae have attracted growing attention in this field due to their unique photosynthetic ability, oxygen-generating capacity, and inherent bioactivities, which can enhance tissue regeneration and angiogenesis. This review systematically outlines the types of microalgae used in bone-related 3D bioprinting, their integration into bioinks and scaffolds, and the mechanisms through which they contribute to bone healing. Compared with previous literature, this review highlights underexplored dimensions such as the immunoregulatory properties of algal-derived scaffolds, the strategies to overcome their limited mechanical strength and immunogenicity<strong>,</strong> and the potential of genetic and material engineering to enhance clinical applicability. Furthermore, we discuss recent advancements in algal-laden scaffold sterilization methods—a critical but often overlooked aspect for clinical translation. A phenotypic–mechanistic integration framework and visual workflow diagrams are provided to aid material selection and design. Finally, we identify current limitations, research gaps, and future directions for utilizing microalgae as functional components in next-generation bioinks. These insights aim to promote the rational design of photosynthetically active and regenerative scaffolds for bone and soft tissue repair.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"408 ","pages":"Pages 121-131"},"PeriodicalIF":3.9,"publicationDate":"2025-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144955285","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Molecular engineering of UnaG: Insights into fluorescence dynamics and biosensor potential UnaG的分子工程：荧光动力学和生物传感器潜力的见解

IF 3.9 2区生物学

Journal of biotechnology Pub Date : 2025-08-28 DOI: 10.1016/j.jbiotec.2025.08.014

Yakup Ulusu , Ramazan Bayat

{"title":"Molecular engineering of UnaG: Insights into fluorescence dynamics and biosensor potential","authors":"Yakup Ulusu , Ramazan Bayat","doi":"10.1016/j.jbiotec.2025.08.014","DOIUrl":"10.1016/j.jbiotec.2025.08.014","url":null,"abstract":"<div><div>UnaG represents the first ligand-inducible (unconjugated bilirubin) fluorescent protein to be discovered in vertebrates. It was isolated from the muscles of the Japanese eel (<em>Anguilla japonica</em>) and has been shown to emit green fluorescence exclusively when bound to unconjugated bilirubin (UC-BR). This distinctive property of UnaG renders it a promising biosensor candidate for the detection of UC-BR. The sensitive and specific detection of UC-BR is of great importance for clinical diagnosis and treatment. It is imperative to enhance the binding affinity and thermal stability of wild type UnaG for successful clinical applications These proteins may serve as promising candidates for future diagnostic applications, pending validation in biological samples. In this study, Arginine at position R112 and R132 in the UnaG-UC-BR binding site was replaced with Methionine. This was done to regulate the binding affinity of wild-type UnaG protein to UC-BR to a level that could be measured in a physiological context, and to improve its thermal stability. In this study, the positive charge in the relevant binding sites was eliminated and the biophysical changes of these mutations on the UnaG protein were investigated. The secondary structures and thermal melting temperatures of the proteins were analysed by circular dichroism spectroscopy. Excitation and emission wavelengths were determined by fluorescence spectroscopy, while dissociation constants (Kd) were determined by titration studies with UC-BR. The findings indicate that R112M, R132M and R112&132 M mutants exhibited high sensitivity, reaching fluorescence saturation at UC-BR concentrations of ∼10 nM, ∼12 nM and ∼10 nM, respectively. Thermal stability analyses revealed that the R112M&R132M double mutant was the most stable variant, with a melting temperature of 72°C. The results of this study indicate that engineered UnaG variants have the potential to function as high-affinity and stable biosensors for the detection of UC-BR.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"407 ","pages":"Pages 87-96"},"PeriodicalIF":3.9,"publicationDate":"2025-08-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144916484","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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