New biotechnology最新文献

{"title":"Screening and bioreactor cultivation of wild-type lactic acid bacteria for high purity D-lactic acid production","authors":"Laura Troiani ,&nbsp;Alessia Levante ,&nbsp;Hannes Russmayer ,&nbsp;Hans Marx ,&nbsp;Erasmo Neviani ,&nbsp;Valentina Bernini ,&nbsp;Camilla Lazzi ,&nbsp;Michael Sauer","doi":"10.1016/j.nbt.2025.08.005","DOIUrl":"10.1016/j.nbt.2025.08.005","url":null,"abstract":"<div><div>Biodegradable polymers are the green alternative to conventional oil-based plastics and have a key role in the achievement of the Sustainable Development Goals of Agenda 2030. Nowadays, polylactic acid (PLA) is one of the most common bioplastics present in the global market. Optically pure D-lactic acid (D-LA) is a valuable monomer for the production of high-performance PLA materials. This study aims to select wild-type lactic acid bacteria capable of producing D-LA with high optical purity and to define an optimized fermentation process in bioreactors to maximize production. A total of 150 LAB strains from the University of Parma Culture Collection (UPCCO) were screened using enzymatic assays and HPLC analysis. Among them, <em>Lactobacillus delbrueckii</em> UPCCO 2214 and <em>Leuconostoc citreum</em> UPCCO 4516 were selected for fed-batch cultivations for their high D-LA production and purity (&gt;97 %). After optimization of fermentation parameters, <em>L. delbrueckii</em> UPCCO 2214 was found to be the most efficient strain for D-LA production, with a yield of 0.74 g/g and a volumetric productivity of 0.96 g/L/h, outperforming <em>L. citreum</em> UPCCO 4516. Bioreactors cultivation has helped to understand microbial production and boost their potentialities. This work supports further investigations and improvements in D-LA production processes to advance the field of biomaterials with benefits for both industry and the environment.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"90 ","pages":"Pages 48-55"},"PeriodicalIF":4.9,"publicationDate":"2025-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144894877","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}

{"title":"Monoclonal antibodies production in microbial systems: Current status, challenges and perspectives","authors":"Di Zhang,&nbsp;Hong Liu,&nbsp;Yaohua Zhong","doi":"10.1016/j.nbt.2025.10.005","DOIUrl":"10.1016/j.nbt.2025.10.005","url":null,"abstract":"<div><div>Monoclonal antibodies (mAbs) serve as indispensable tools in diagnostics, clinical therapeutics, and biomedical research. However, their large-scale production faces significant challenges due to the high costs and lengthy timelines associated with conventional mammalian cell-based expression systems. Microbial expression platforms have emerged as a transformative alternative, offering cost-effectiveness, rapid cultivation cycles, and superior genetic tractability for industrial-scale monoclonal antibodies production. Recent advances in Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9-mediated gene editing enable precise metabolic engineering of host strains to enhance protein folding, secretion efficiency, and translational accuracy. Synthetic biology approaches facilities the reconstruction of mammalian glycosylation pathways in microbial systems, yielding monoclonal antibodies with near-native structural integrity. Furthermore, AI (artificial intelligence)-driven optimization of expression vectors, promoter systems, and culture conditions, combined with high-throughput screening of engineered strains, significantly accelerates the identification of high-yield production clones. This review comprehensively examines current progress in microbial expression systems, strain engineering strategies, and fermentation optimization for enhanced monoclonal antibodies production, while critically discussing existing limitations and potential solutions to advance the field.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"90 ","pages":"Pages 163-173"},"PeriodicalIF":4.9,"publicationDate":"2025-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145266027","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}

{"title":"The Filterprep: A simple and efficient approach for high-yield, high-purity plasmid DNA purification","authors":"Yu-Hsuan Cheng ,&nbsp;Yu-Jiu Wu ,&nbsp;Yung-Chun Shih ,&nbsp;Yu-Qian Lin ,&nbsp;Yu-Wei Chang ,&nbsp;Yu-Heng Wu ,&nbsp;En-Wei Hu ,&nbsp;Ren-Hsuan Ku ,&nbsp;Cheng-Mu Wu ,&nbsp;Shao-Chi Wu ,&nbsp;Ting-Yu Yeh ,&nbsp;Chung-Te Chang","doi":"10.1016/j.nbt.2025.09.003","DOIUrl":"10.1016/j.nbt.2025.09.003","url":null,"abstract":"<div><div>Plasmid DNA purification remains a fundamental yet resource-intensive step in molecular biology and biotechnology. Conventional protocols often yield plasmids with suboptimal purity, while commercial kits, though efficient, are costly and use proprietary formulations with limited transparency for troubleshooting or customization. Here, we present Filterprep, a hybrid method combining classical ethanol precipitation with a single spin-column cleanup step, enabling rapid recovery of high-purity plasmid DNA with yields notably higher than those obtained using commercial kits, in approximately 40 min, with yields up to 5-fold greater than those of representative commercial midiprep kits under matched conditions. Filterprep offers a transparent, cost-effective, and scalable alternative, simplifying workflows and reducing hands-on time without compromising DNA quality. The purified plasmids are compatible with a variety of downstream applications, including mammalian cell transfection, protein expression, and reporter assays, while remaining stable for 12 months at −20 °C. With its balance of efficiency and practicality, Filterprep is well suited for high-throughput laboratories and resource-limited settings.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"90 ","pages":"Pages 77-87"},"PeriodicalIF":4.9,"publicationDate":"2025-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145060003","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}

{"title":"Characterization and orthogonality assessment of two quorum sensing systems for synthetic biology applications","authors":"Jasmine De Baets,&nbsp;Brecht De Paepe,&nbsp;Marjan De Mey","doi":"10.1016/j.nbt.2025.08.001","DOIUrl":"10.1016/j.nbt.2025.08.001","url":null,"abstract":"<div><div>Quorum sensing systems have a broad range of applications within the field of synthetic biology. However, a bottleneck is the optimization and tuning of these systems due to the lack of standardization and complete characterization. In this research, two quorum sensing systems, namely the LasI/LasR and the EsaI/EsaR system, were fully characterized in the model host organism <em>Escherichia coli</em>. Furthermore, insight was gained in the interplay between the various parts of these systems. To further expand the range of possibilities with these quorum sensing systems, the orthogonality of the two systems was assessed to allow simultaneous use within the same cell without interfering crosstalk. This assessment was performed on three levels: promoter, signal and synthase crosstalk. It was demonstrated that LasR is able to interact with the promoter of the EsaI/EsaR system, albeit to a low extent. Additionally, LasR was able to respond to the autoinducers produced by EsaI. To solve the promoter crosstalk, a nucleotide change was introduced into the binding site of EsaR within the promoter region. Additionally, LasR mutants were created rationally and screened for decreased response to EsaI while retaining functionality. The best performing mutant, LasR(P117S), was further characterized. In conclusion, we have further unlocked the potential of quorum sensing systems for synthetic biology applications by obtaining two functional, characterized and orthogonal quorum sensing systems.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"90 ","pages":"Pages 20-35"},"PeriodicalIF":4.9,"publicationDate":"2025-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144886643","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}

{"title":"Effect of salinity on the composition of a seawater-adapted strain of Scenedesmus almeriensis","authors":"Elia Rivera-Sánchez ,&nbsp;Silvia Villaró-Cos ,&nbsp;María Salinas-García ,&nbsp;Francisco Javier Alarcón ,&nbsp;Marco García-Vaquero ,&nbsp;Lucie K. Tintrop ,&nbsp;Daniel Kurpan ,&nbsp;Tomás Lafarga","doi":"10.1016/j.nbt.2025.09.008","DOIUrl":"10.1016/j.nbt.2025.09.008","url":null,"abstract":"<div><div>The main objective of this study was to evaluate the potential adaptation to seawater of the freshwater strain <em>Scenedesmus almeriensis</em> and to evaluate the effect of salinity on its growth, morphology, and biochemical composition. Incorporating low seawater concentrations into the culture medium (up to 103 mM NaCl) resulted in an increase in biomass productivity from 0.15 to 0.22 g·L^-¹ ·day^-¹ and an increase in the maximum specific growth rate from 0.14 to 0.26 day^-¹. This was attributed to the presence of micronutrients in the seawater and adaptive responses to stress. Despite a lower biomass productivity (0.11 g·L⁻¹·day^-¹), <em>Scenedesmus almeriensis</em> was able to grow well (0.15 day⁻¹) in a medium formulated with only seawater and commercial fertilisers. Cell morphology was significantly affected, with a 150 % increase in cell perimeter and an increase in roundness from 61.5 % (freshwater) to 95.8 % (seawater). The use of seawater also affected the chemical composition of the biomass. Seawater favoured the synthesis of specific fatty acids that have nutritional and industrial value, including polyunsaturated fatty acids. The protein content was slightly reduced under saline conditions but remained at 40 % (which is high compared to other biomasses). The results highlight the potential of seawater as a sustainable and cost-effective substitute for freshwater to produce <em>Scenedesmus almeriensis</em>. Future studies will validate the production of these strains at the large scale and identify potential industrial uses for the biomass produced.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"90 ","pages":"Pages 114-121"},"PeriodicalIF":4.9,"publicationDate":"2025-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145154160","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}

{"title":"The diversity and keystone species of cucumber rhizosphere microbiome: Unveiling their role in driving cucumber growth and microbial communities","authors":"Yaping Yin ,&nbsp;Raja Asad Ali Khan ,&nbsp;Liping Li ,&nbsp;Yinggu Wu ,&nbsp;Chun Li ,&nbsp;Yuejian Li ,&nbsp;Sen Ren ,&nbsp;Jing Zhang ,&nbsp;Weiwei Wang ,&nbsp;Runmao Lin ,&nbsp;Manman Zhang ,&nbsp;Xiaojun Liu ,&nbsp;Jumei Hou ,&nbsp;Genyun Liang","doi":"10.1016/j.nbt.2025.09.007","DOIUrl":"10.1016/j.nbt.2025.09.007","url":null,"abstract":"<div><h3>Background</h3><div>Studying the interrelationship between agroecosystems and the microbiome is essential in achieving sustainable agricultural development. In particular, the relationship between the rhizosphere microbiome and crops is critical to the health and stability of agroecosystems. This study evaluated the diversity and keystone species of the cucumber rhizosphere microbiome and investigated their contributing role in cucumber growth and microbial community regulation. A total of 108 soil samples were collected from the rhizosphere and bulk soil of cucumber crops in different locations and under different cultivation modes in a high-yield area. These samples were analyzed to study the interrelationships between different cultivation modes and crop rhizosphere microorganisms.</div></div><div><h3>Results</h3><div>The results revealed significant regional differences in microbial communities due to microhabitat (rhizosphere and bulk soil) and geographic location. Common bacteria such as <em>Pseudomonas</em>, <em>Sphingomonas</em>, <em>Lysobacter</em> and <em>Rhodanobacter</em> were the most abundant and showed strong correlations with several other bacteria through co-occurrence networking. In addition, <em>Flavobacterium</em> sp. and <em>Bacillus</em> sp. were identified as keystone species for the rhizobia community structure. Synthetic microbial communities and pot experiments confirmed that these two keystone species could enrich the soil microbime and promote cucumber growth. <em>Flavobacterium</em> sp. may promote cucumber growth by regulating the rhizosphere microbial community while <em>Bacillus</em> sp. may influence cucumber growth by stabilizing the rhizosphere microbial biomass, further enhancing nutrient cycling efficiency and improving system stability, which in turn affected the agroecosystem.</div></div><div><h3>Conclusions</h3><div>Our study demonstrates that specific keystone species are important in shaping the cucumber rhizosphere microbiome and plant growth. These results provide insights into agroecosystem-microbe interrelationships and provide a potential basis for optimizing agricultural management strategies.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"90 ","pages":"Pages 134-144"},"PeriodicalIF":4.9,"publicationDate":"2025-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145154725","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}

{"title":"Prenylation of diverse indole derivatives by the fungal aromatic prenyltransferase RePT","authors":"Pimvisuth Chunkrua,&nbsp;Mirjam A. Kabel,&nbsp;Jean-Paul Vincken,&nbsp;Willem J.H. van Berkel,&nbsp;Wouter J.C. de Bruijn","doi":"10.1016/j.nbt.2025.09.002","DOIUrl":"10.1016/j.nbt.2025.09.002","url":null,"abstract":"<div><div>Prenylation is a widespread natural modification of compounds that serves to functionalize and often enhance the bioactivity of plant and microbial secondary metabolites, including indole derivatives. In this study, we aimed to expand the library of prenylated indoles using RePT, a fungal (i.e. <em>Rasamsonia emersonii</em>) aromatic prenyltransferase from the dimethylallyl tryptophan synthase (DMATS) family. Previous work showed that RePT readily <em>C</em>7- and <em>N</em>1-prenylated <span>l</span>-tryptophan, and <em>O</em>-prenylated <span>l</span>-tyrosine and a number of phenolic stilbenes. Here, we investigated its regioselectivity further with 23 indole substrates, including tryptophan derivatives with varying <em>C</em>4-<em>C</em>7 substituents and several <em>C</em>3-substituted indoles. High conversion was observed primarily with fluorinated tryptophans and unsubstituted indole. Product analysis by UHPLC-PDA-ESI-MSⁿ and NMR revealed that RePT mainly catalyzed either normal prenylation at <em>C</em>7 or reverse prenylation at <em>N</em>1 on a series of halogenated tryptophans. The regioselectivity observed for several substrates was strongly influenced by the position of the halogen substituent, particularly fluorine, which displayed its characteristic <em>ortho-</em>/<em>para-</em>directing effect. In the absence of the amino acid moiety, RePT’s regioselectivity in some cases shifted from its typical preference, leading to prenylation at alternative positions such as <em>C</em>3 and <em>C</em>6. These findings showcase the versatility of RePT for modifying diverse indole derivatives and demonstrate, for the first time, halogen-induced steering of the regioselectivity of DMATS to facilitate synthesis of bioactive prenylated compounds and intermediates.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"90 ","pages":"Pages 88-96"},"PeriodicalIF":4.9,"publicationDate":"2025-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145086626","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}

{"title":"Site-specifically immobilized D-amino acid dehydrogenase for the synthesis of D-phenylalanine","authors":"Krisztina Boros ,&nbsp;András-Ernő Iszlai ,&nbsp;Laura-Edit Barabás,&nbsp;László Csaba Bencze,&nbsp;Monica Ioana Toşa","doi":"10.1016/j.nbt.2025.09.006","DOIUrl":"10.1016/j.nbt.2025.09.006","url":null,"abstract":"<div><div>Aromatic <span>d</span>-amino acids (<span>d</span>-AAs) have gained increasing attention as chiral building blocks, with biocatalytic procedures emerging as powerful methods for their asymmetric synthesis. <span>d</span>-Amino acid dehydrogenases (<span>D</span>AADH), developed by protein engineering from <em>meso</em>-diaminopimelate dehydrogenases, step out as highly efficient biocatalysts for the reductive amination-based production of <span>d</span>-AAs. Enzyme immobilization allows the recovery and reuse of biocatalysts, while also enhances their operational stability, which is essential for industrial applications. Since the immobilization of DAADHs have been less explored, we targeted to further progress within the covalent immobilization of DAADH from <em>Ureibacillus thermosphaericus</em>, by exploring its site-specific, covalent immobilization. The individual replacement of several surficial Ser residues to Cys at positions 2, 58, 92, 185, 192 and 317 of <em>Ut</em>DAADH, allowed their site-specific immobilization onto the maleimide-functionalized Purolite® ECR8415F methacrylic support. The highest specific activity values, <span><math><mo>∼</mo></math></span>0.078 and <span><math><mo>∼</mo></math></span>0.083 U/mg provided by immobilization through Cys2 and Cys192, respectively, showed 2.1- and 2.2-fold higher values compared to the covalently, but non-specifically immobilized <em>Ut</em>DAADH. The recyclability of the immobilized preparations was tested among 10 reductive amination-cycles of phenylpyruvate and based on the retained conversion and specific activities, <em>Ut</em>DAADH immobilized through S2C was the best-performing biocatalyst, maintaining 65–70 % conversion and 50 % of the initial activity after the 10^th cycle. After the assessment of optimal enzyme/substrate <em>ratio</em>, the immobilized S2C <em>Ut</em>DAADH was tested in three consecutive 200 mg-scale reaction, providing the enantiopure <span>d</span>-Phe with complete conversions and excellent &gt; 88 % isolation yields, supporting its synthetic applicability.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"90 ","pages":"Pages 145-154"},"PeriodicalIF":4.9,"publicationDate":"2025-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145213212","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}

{"title":"Comparative transcriptomics of CAZy enzymes in white- and brown-rot agaricomycetes: Evolutionary insights into lignocellulose degradation and the relevance of GH16 glycoside hydrolase functional divergence","authors":"Idoia Jiménez ,&nbsp;Manuel Alfaro ,&nbsp;Edurne Garde ,&nbsp;Gumer Pérez ,&nbsp;Ana Fernandez-Morales ,&nbsp;Anna Lipzen ,&nbsp;Kathleen Lail ,&nbsp;Diane Bauer ,&nbsp;Kerrie Barry ,&nbsp;Igor V. Grigoriev ,&nbsp;Lucía Ramírez ,&nbsp;Antonio G. Pisabarro","doi":"10.1016/j.nbt.2025.10.004","DOIUrl":"10.1016/j.nbt.2025.10.004","url":null,"abstract":"<div><div>White-rot and brown-rot fungi (WRF and BRF, respectively) decompose lignocellulose, the main structural component of plant biomass, through distinct mechanisms. This study examines the transcriptomic responses of three WRF species (<em>Pleurotus ostreatus</em>, <em>Phanerochaete chrysosporium</em>, and <em>Heterobasidion irregulare</em>) and two BRF species (<em>Fomitopsis schrenkii</em> and <em>Rhodonia placenta</em>) grown on poplar wood (W) and glucose (G) as sole carbon sources. RNA-seq analysis revealed upregulation of carbohydrate-active enzymes (CAZymes) linked to lignocellulose degradation. WRF displayed a broader enzymatic repertoire, whereas BRF employed a more selective approach. Among these responses, GH16 glycoside hydrolases were consistently upregulated across all species, including BRF. Since GH16 enzymes are involved in both plant hemicellulose modification and cell wall remodeling, their wood-induced expression may reflect multiple processes rather than a single conserved wood-decay mechanism. Structural and phylogenetic analyses revealed species-specific divergence, consistent with potential functional specialization. These findings broaden our understanding of fungal enzymatic strategies and highlight GH16 enzymes as candidates for further study. Beyond the immediate context of wood decay, this work has broader implications for fungal ecology, evolutionary biology, and biotechnological applications such as biomass conversion and sustainable bioenergy.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"90 ","pages":"Pages 236-256"},"PeriodicalIF":4.9,"publicationDate":"2025-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145280736","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}

{"title":"Using control bias to identify initial targets for bioproduction improvement","authors":"Michael Binns ,&nbsp;Pedro de Atauri ,&nbsp;Marta Cascante ,&nbsp;Constantinos Theodoropoulos","doi":"10.1016/j.nbt.2025.07.008","DOIUrl":"10.1016/j.nbt.2025.07.008","url":null,"abstract":"<div><div>Sensitivity analysis of bioprocess metabolic reaction networks analysis allows the prediction of system parameters such as those associated with the enzyme activity of certain reaction steps which significantly affect the overall production. However, uncertainties in kinetic rate expressions and in the resulting steady-state flux distributions limit the accuracy of these predictions. Starting from minimal information (reaction stoichiometry, and external fluxes in/out of the system and potentially identification of steps at equilibrium) a new preliminary method is proposed using sampling of elasticities and metabolic fluxes to calculate the control bias. The calculated control bias identifies steps which are likely to have positive control, negative control or negligible/uncertain control. This is intended to give initial guidance before further detailed investigation is carried out, identifying targets for any organism to enhance production of valuable chemicals. As a case study, this methodology is applied to succinic acid bioproduction using <em>Actinobacillus succinogenes</em> and analysis successfully reveals the reaction steps having the greatest positive and negative influence on biosuccinic acid production.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"89 ","pages":"Pages 130-140"},"PeriodicalIF":4.9,"publicationDate":"2025-11-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144718160","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}