New biotechnology最新文献

Phenolic compounds fingerprints show distinct ligninolytic bacteria behaviours during technical lignins conversion 酚类化合物指纹在木质素技术转化过程中显示出不同的木质素分解细菌行为

IF 4.5 2区生物学

New biotechnology Pub Date : 2025-06-28 DOI: 10.1016/j.nbt.2025.06.004

Corinne Ivaldi , Brigitte Chabbert , Roland Molinié , David Crônier , David Mathiron , Jean Xavier Fontaine , François Mesnard , Frédéric Velard , Harivony Rakotoarivonina

{"title":"Phenolic compounds fingerprints show distinct ligninolytic bacteria behaviours during technical lignins conversion","authors":"Corinne Ivaldi , Brigitte Chabbert , Roland Molinié , David Crônier , David Mathiron , Jean Xavier Fontaine , François Mesnard , Frédéric Velard , Harivony Rakotoarivonina","doi":"10.1016/j.nbt.2025.06.004","DOIUrl":"10.1016/j.nbt.2025.06.004","url":null,"abstract":"<div><div>Lignins, one of the main components of plant cell wall, and by-products of certain industries (paper and wood industries,…) are a renewable source of aromatic molecules. They can be degraded and transformed by microbial and enzymatic processes known to be respectful of the environment. Biological valorization of lignins remains challenging as biocatalysts are not sufficiently effective and efficient. Moreover, the chemical complexity and heterogeneity of lignins are a barrier to their use. Understanding the microbial behaviour on lignins by fingerprinting their efficient transformation could lead to the development of effective biological routes to valorise these aromatic polymers. Ligninolytic bacteria present some interesting features in term of ligninolytic enzymes productions, utilization of aromatic compounds <em>via</em> various intracellular pathways and the productions of molecules of interest from aromatic molecules. In this work, multiple approaches (growth studies, ligninolytic activities production, lignin modifications and phenolic compounds fingerprints) were used to understand the behaviour of two ligninolytic bacteria <em>Pandoraea norimbergensis</em> and <em>Comamonas composti</em>, in presence of lignins with different structures and origins. Results showed dissimilar growths profiles, lignin modifications, consumption and production of phenolic monomers and oligomers according to the bacteria and the lignin used. To achieve efficient transformation of lignins, suitable combination of biocatalysts and lignins is required and the microorganisms used must be selected on the basis of their metabolic capacity, and the structure and composition of lignin.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"89 ","pages":"Pages 60-72"},"PeriodicalIF":4.5,"publicationDate":"2025-06-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144513731","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

Single-cell proteins polyhydroxyalkanoates-rich microbial biomass from municipal and winery waste as potential additive for aquafeeds 从城市和酿酒厂废料中提取的富含多羟基烷酸盐的微生物生物量作为水产饲料的潜在添加剂

IF 4.5 2区生物学

New biotechnology Pub Date : 2025-06-21 DOI: 10.1016/j.nbt.2025.06.003

Giulia Adele Tuci , Marco Gottardo , Aditi Parmar Chitharanjan , Gloriana Cardinaletti , Giulia Pascon , Paolo Pavan , Francesco Valentino

{"title":"Single-cell proteins polyhydroxyalkanoates-rich microbial biomass from municipal and winery waste as potential additive for aquafeeds","authors":"Giulia Adele Tuci , Marco Gottardo , Aditi Parmar Chitharanjan , Gloriana Cardinaletti , Giulia Pascon , Paolo Pavan , Francesco Valentino","doi":"10.1016/j.nbt.2025.06.003","DOIUrl":"10.1016/j.nbt.2025.06.003","url":null,"abstract":"<div><div>This study evaluated single-cell protein production from PHA-rich mixed microbial cultures obtained from fermentation and subsequent PHA storage, using urban (namely food waste and municipal sewage sludge; FW-MSS) and agricultural waste (namely wine lees; WL) streams as substrates. FW-MSS fermentation achieved stable short-chain fatty acid (SCFA) production and a high COD<sub>SCFA</sub>/COD<sub>SOL</sub> ratio of 0.77 ± 0.01, which allowed to select a mixed microbial culture (MMC) with intracellular PHA content of 15.1 wt%, which aligns with fish dietary standards and yielded a MMC biomass with a protein level of 55.1 wt% and a balanced essential amino acid (EAA) profile. In contrast, WL fermentation showed lower SCFA content and stability, yielding a MMC with 45.8 wt% of protein along with a high non-conformance rate (53.65 %), and 7.2 wt% PHA, making the resulting MMC more suited as a supplemental protein source. Distinct microbial communities developed in the two SBRs due to different feedstocks, influencing the abundance of PHA-storing bacteria, with no known fish pathogens detected in either sample. Statistical analysis confirmed FW-MSS’s superior product consistency, supporting its potential as a good quality SCP for aquafeed, especially for rainbow trout, as confirmed by its high essential amino acid index (EAAI).</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"89 ","pages":"Pages 29-39"},"PeriodicalIF":4.5,"publicationDate":"2025-06-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144366897","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

On-demand insulin manufacturing using cell-free systems with an "on-column" conversion approach 按需胰岛素制造使用无细胞系统与“柱上”转换方法。

IF 4.5 2区生物学

New biotechnology Pub Date : 2025-06-20 DOI: 10.1016/j.nbt.2025.06.002

Shayan G. Borhani , Max Z. Levine , Chandrasekhar Gurramkonda , Yanyan Qu , Dingyin Tao , Christopher A. LeClair , James R. Swartz , Govind Rao

{"title":"On-demand insulin manufacturing using cell-free systems with an \"on-column\" conversion approach","authors":"Shayan G. Borhani , Max Z. Levine , Chandrasekhar Gurramkonda , Yanyan Qu , Dingyin Tao , Christopher A. LeClair , James R. Swartz , Govind Rao","doi":"10.1016/j.nbt.2025.06.002","DOIUrl":"10.1016/j.nbt.2025.06.002","url":null,"abstract":"<div><div>Recent studies project that the prevalence of diabetes is expected to increase significantly and lead to escalating demand on the insulin supply chain. Despite being the first recombinant therapeutic approved by the FDA, insulin remains challenging to access for many around the globe. Here we report on advancements in manufacturing insulin using cell-free protein synthesis (CFPS) systems to rapidly produce mature desB30-insulin in less than a day. This is a major advance towards decentralizing insulin manufacturing and bringing production closer to the point-of-care, thereby improving diabetic patient accessibility. To this end, a purified cell-free extract, PUREfrex® 2.1, was utilized to synthesize a tagged proinsulin construct that can be captured and converted into mature insulin using an on-column affinity chromatography process. Notably, two chaperones, peptidyl prolyl isomerase (FkpA), and seven kilodalton protein (Skp) were observed to play a critical role during cell-free expression of proinsulin. The proinsulin was then immobilized on a Ni-NTA column where the purification and conversion of cell-free products were performed sequentially to yield desB30-insulin. Following further optimization, this method serves as a time and resource-efficient production process as compared to current methods. When applied simultaneously, the cell-free expression and on-column conversion methods reported here can be adopted to enable rapid on-demand insulin manufacturing in order to improve the accessibility of insulin and prevent future shortages.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"89 ","pages":"Pages 51-59"},"PeriodicalIF":4.5,"publicationDate":"2025-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144369106","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

Improving PHB production in cyanobacteria: Modeling the optimal light regime for growth. 提高蓝藻中PHB的生产：模拟生长的最佳光照制度。

IF 4.5 2区生物学

New biotechnology Pub Date : 2025-06-17 DOI: 10.1016/j.nbt.2025.05.005

I Perez Counago, J M Fernandez Montenegro, S Iglesias Moreira, F Rodriguez Lorenzo, M Placer Lorenzo, P Villar Sola, E Pancorbo González, J Illade Quinteiro, L Herrero Castilla, J A Álvarez Rodríguez, B Altamira Algarra, E Gonzalez Flo, J Garcia, F Guedes, M Lopez-Garcia, S Muiños-Landin

{"title":"Improving PHB production in cyanobacteria: Modeling the optimal light regime for growth.","authors":"I Perez Counago, J M Fernandez Montenegro, S Iglesias Moreira, F Rodriguez Lorenzo, M Placer Lorenzo, P Villar Sola, E Pancorbo González, J Illade Quinteiro, L Herrero Castilla, J A Álvarez Rodríguez, B Altamira Algarra, E Gonzalez Flo, J Garcia, F Guedes, M Lopez-Garcia, S Muiños-Landin","doi":"10.1016/j.nbt.2025.05.005","DOIUrl":"https://doi.org/10.1016/j.nbt.2025.05.005","url":null,"abstract":"<p><p>The production of bioplastics, such as polyhydroxybutyrate (PHB), using cyanobacteria offers a sustainable alternative to conventional plastics. However, achieving economically viable production requires optimizing biomass growth. This study examined four growth models: Gompertz (empirical growth), Baranyi-Roberts (biologically dependent), Monod (nutrient dependent), and Aiba (irradiance dependent). The results indicate that a light-based model more accurately describes cyanobacterial growth and shows potential for optimizing light regimes. Additionally, an estimator was proposed to assess the potential PHB yield within the given biomass. Experiments were conducted to correlate photosynthetic efficiency with biomass production, providing deeper insights into the effects of light on growth. These findings support the development of optimized cultivation strategies, ultimately improving the economic viability of cyanobacteria-based bioplastics.</p>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":" ","pages":""},"PeriodicalIF":4.5,"publicationDate":"2025-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144485268","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

Effect of operating conditions on the microbial selection of P(3HB-co-3HV) producers in open continuous culture 开放式连续培养中操作条件对P（3HB-co-3HV）生产者微生物选择的影响

IF 4.5 2区生物学

New biotechnology Pub Date : 2025-06-13 DOI: 10.1016/j.nbt.2025.06.001

Matteo Castiello , Laetitia Cavaillé , Mansour Bounouba , Simon Dubos , Claire Dumas , Etienne Paul

{"title":"Effect of operating conditions on the microbial selection of P(3HB-co-3HV) producers in open continuous culture","authors":"Matteo Castiello , Laetitia Cavaillé , Mansour Bounouba , Simon Dubos , Claire Dumas , Etienne Paul","doi":"10.1016/j.nbt.2025.06.001","DOIUrl":"10.1016/j.nbt.2025.06.001","url":null,"abstract":"<div><div>From the perspective of producing P(3HB-co-3HV), the selection of PHA accumulating organisms from activated sludge with a feeding incorporating propionate was investigated in open continuous culture under dual carbon and phosphorus limitation conditions. The selected consortia were then harvested in batch cultures with phosphorus (P) deficiency to evaluate their PHA storage potential. Microbial selection exhibited consistent functional stability of PHA production, emphasizing the robustness of this enrichment strategy. A comparison with previous studies revealed that at a given degree of P limitation, the distribution of carbon consumption between cell growth, PHA storage and maintenance reactions was conditioned by the nature of the carbon source. The combination of the nature of the carbon substrate, the degree of P limitation and the dilution rate governs microbial competition. The dominant bacterial genera showed very heterogeneous PHA production capacities. <em>Acinetobacte</em>r spp., whose establishment was favored by the presence of acetate, a high dilution rate, and a low C/P ratio, was found to be a poor producer. In contrast, <em>Malikia</em> spp. and <em>Zoogloea</em> spp., mostly selected with butyrate, a lower dilution rate, and strong P limitation, exhibited high specific PHA production rates.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"89 ","pages":"Pages 40-50"},"PeriodicalIF":4.5,"publicationDate":"2025-06-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144302586","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

Tailored enzyme expression modifies Shewanella oneidensis biofilms and increases current density 量身定制的酶表达修饰希瓦氏菌生物膜和增加电流密度

IF 4.5 2区生物学

New biotechnology Pub Date : 2025-06-09 DOI: 10.1016/j.nbt.2025.05.006

Edina Marlen Klein , Hannah Heintz , René Wurst , Christian Jonas Lapp , Miriam Edel , Hendrik Hähl , Karin Jacobs , Johannes Gescher

{"title":"Tailored enzyme expression modifies Shewanella oneidensis biofilms and increases current density","authors":"Edina Marlen Klein , Hannah Heintz , René Wurst , Christian Jonas Lapp , Miriam Edel , Hendrik Hähl , Karin Jacobs , Johannes Gescher","doi":"10.1016/j.nbt.2025.05.006","DOIUrl":"10.1016/j.nbt.2025.05.006","url":null,"abstract":"<div><div>Biofilm formation is the most effective pathway for electron transfer to anodes in bioelectrochemical systems. However, the mechanisms triggering biofilm formation under anoxic conditions, as well as the architectural and compositional factors that positively influence current generation, are not well understood. Recent findings have shown that riboflavin can function similarly to a quorum sensing molecule in the γ-proteobacterium <em>Shewanella oneidensis</em>. Enhanced biofilm formation induced by riboflavin correlates with increased current densities. Only a limited number of candidate proteins were found to have altered concentrations due to this quorum sensing mechanism. This study demonstrates that the catalytic functions of the UDP-N-acetylglucosamine C4 epimerase WbpP and UDP-N-acetyl-D-glucosamine 6-dehydrogenase WbpA affect biofilm formation and lead to increased current density. Using optical coherence tomography, we found that the expression of each protein individually causes specific, quantifiable changes in biofilm architecture, including biovolume, height, and porosity. However, the current density did not significantly differ when these proteins were expressed alone compared to the control. In contrast, co-expression of WbpP and WbpA resulted in a doubling of current density, closely resembling the increases observed with riboflavin-mediated quorum sensing. We hypothesize that riboflavin-based quorum sensing may lead, through several intermediary steps, to the overproduction of WbpA and WbpP, resulting in better attachment to graphite anodes and thus higher current densities.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"89 ","pages":"Pages 1-10"},"PeriodicalIF":4.5,"publicationDate":"2025-06-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144263229","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

Sprinkling in extra validation for high-value PTMs and therapeutic Abs with MILKSHAKE Western blots and Sundae ELISAs 用MILKSHAKE Western blots和Sundae elisa对高价值PTMs和治疗性抗体进行额外验证。

IF 4.5 2区生物学

New biotechnology Pub Date : 2025-06-06 DOI: 10.1016/j.nbt.2025.05.007

Qiana Mendez , Srinivas S. Thota , Jason J. Oliveira , Hannah J. Crary , Suchetana Saha , Kezzia S. Jones , Lourdes Perez , Felicity Acca , Cassandra D. Chapados , Holland A. Driscoll , Xiaofeng Li , Gregory R. Mirando , Mikhail A. Kostylev , Erik C. Gunther , Brian K. Kay , Michael P. Weiner , Mary R. Ferguson

{"title":"Sprinkling in extra validation for high-value PTMs and therapeutic Abs with MILKSHAKE Western blots and Sundae ELISAs","authors":"Qiana Mendez , Srinivas S. Thota , Jason J. Oliveira , Hannah J. Crary , Suchetana Saha , Kezzia S. Jones , Lourdes Perez , Felicity Acca , Cassandra D. Chapados , Holland A. Driscoll , Xiaofeng Li , Gregory R. Mirando , Mikhail A. Kostylev , Erik C. Gunther , Brian K. Kay , Michael P. Weiner , Mary R. Ferguson","doi":"10.1016/j.nbt.2025.05.007","DOIUrl":"10.1016/j.nbt.2025.05.007","url":null,"abstract":"<div><div>Thoroughly validated antibodies (Abs) are crucial for the generation of meaningful scientific data. Abs for post translationally modified (PTM) protein targets in particular pose added validation challenges. The MILKSHAKE method employs surrogate proteins which are either modified or non-modified at a specific site. Western blot is used to observe the binding of PTM Abs to the surrogate proteins, indicating the specificity of the PTM Ab under test. In this study, we expand the utility of MILKSHAKE by validating acetyl and methyl specific Abs and by introducing another surrogate protein antigen based on cellulose binding domain (CBD) to evaluate Abs in a single western blot lane. This study also explores the use of ‘Sundae’ surrogate protein ELISA specifically for therapeutic Ab evaluation.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"89 ","pages":"Pages 11-19"},"PeriodicalIF":4.5,"publicationDate":"2025-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144248903","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

Microbial synthesis of polyhydroxyalkanoate blends with engineered Pseudomonas putida 聚羟基烷酸酯与工程恶臭假单胞菌共混物的微生物合成。

IF 4.5 2区生物学

New biotechnology Pub Date : 2025-06-02 DOI: 10.1016/j.nbt.2025.05.004

Minglong Li , Khalid Doudin , David B. Robins , Georgios Tetradis-Mairis , Tuck Seng Wong , Kang Lan Tee

{"title":"Microbial synthesis of polyhydroxyalkanoate blends with engineered Pseudomonas putida","authors":"Minglong Li , Khalid Doudin , David B. Robins , Georgios Tetradis-Mairis , Tuck Seng Wong , Kang Lan Tee","doi":"10.1016/j.nbt.2025.05.004","DOIUrl":"10.1016/j.nbt.2025.05.004","url":null,"abstract":"<div><div>Polyhydroxyalkanoates (PHAs) are biopolymers naturally produced by various microorganisms and offer a sustainable alternative to fossil fuel-derived plastics. They can be synthesized from diverse feedstock, including waste biomass such as lignocellulose, municipal waste, sludge, and industrial by-products. To tailor their properties for specific applications, PHAs are typically blended post synthesis. An alternative approach is the direct synthesis of PHA blends in a single fermentation, which can reduce the need for multiple separate fermentations and extractions. In this study, we engineered <em>Pseudomonas putida</em> to synthesize PHA blends composed of poly-3-hydroxybutyrate [P3(HB)] and medium-chain-length PHA (mcl-PHA). Through using different promoters, blends with 3HB monomer content ranging from 17.9 mol% to 99.6 mol% were produced. Optimizing cultivation conditions yielded a maximum PHA production of 1.48 ± 0.15 g/L, with a PHA content of 52.2 ± 4.3 wt% of cell dry weight. A combination of gel permeation chromatography, nuclear magnetic resonance and diffusion ordered spectroscopy were employed to determine the molecular weight and confirm the identity of the PHA blend, revealing in all cases, a higher molecular weight P(3HB) than mcl-PHA. The blends produced had thermal properties comparable to PHA blends produced by post synthesis melt compounding. This work demonstrates the microbial synthesis of PHA blends in <em>P. putida</em> and is the first instance of blend composition control via promoter selection, paving the way for the one-step biomanufacturing of customizable PHA blends.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"88 ","pages":"Pages 161-170"},"PeriodicalIF":4.5,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144226063","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

Establishment of a cell-free multi-enzyme cascade for the synthesis of UDP-GalNAc 无细胞多酶级联合成UDP-GalNAc的建立。

IF 4.5 2区生物学

New biotechnology Pub Date : 2025-05-30 DOI: 10.1016/j.nbt.2025.05.003

Tuan Son Hoang , Sara-Theres Wormstall , Nam-Hai Hoang , Udo Reichl , Thomas F.T. Rexer

{"title":"Establishment of a cell-free multi-enzyme cascade for the synthesis of UDP-GalNAc","authors":"Tuan Son Hoang , Sara-Theres Wormstall , Nam-Hai Hoang , Udo Reichl , Thomas F.T. Rexer","doi":"10.1016/j.nbt.2025.05.003","DOIUrl":"10.1016/j.nbt.2025.05.003","url":null,"abstract":"<div><div>UDP-<em>N</em>-acetylgalactosamine (UDP-GalNAc) is an essential building block in the synthesis of glycans including <em>O</em>-glycans and glycosaminoglycans. For the latter, enzymatic synthesis is often a promising approach for producing multi-gram amounts. However, the high cost of UDP-GalNAc has limited this approach. This study reports on the development and optimization of a multi-enzyme cascade for synthesizing UDP-GalNAc from inexpensive substrates. Consisting of six recombinant enzymes, the cascade converts uridine (Uri) and GalNAc to UDP-GalNAc with <em>in situ</em> ATP regeneration using polyphosphate (PolyP<sub>n</sub>). Two rounds of Design of Experiments (DoE) optimization were performed to systematically evaluate and optimize reaction parameters including pH, temperature, MgCl<sub>2</sub>, ATP, and PolyP<sub>n</sub> concentrations. The established cascade achieved a percentage yield of 95 % and an UDP-GalNAc titer of 46.1 mM (28 g/L). This represented a 19-fold improvement over the initial conditions. Purification by anion-exchange chromatography yielded a maximum recovery of 89 % and a purity of 90 %. Overall, this scalable, low-cost enzymatic synthesis of UDP-GalNAc overcomes current limitations in availability and cost, potentially enabling new applications in the field of glycobiotechnology.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"89 ","pages":"Pages 20-28"},"PeriodicalIF":4.5,"publicationDate":"2025-05-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144199706","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

MutS-mediated rapid and cost-effective error correction in in vitro DNA synthesis mts介导的体外DNA合成的快速和高成本效益的错误纠正。

IF 4.5 2区生物学

New biotechnology Pub Date : 2025-05-28 DOI: 10.1016/j.nbt.2025.05.002

Liting Jia , Shuang Wang , Dongmei Wang , Xingjiang Li , Jiong Hong

{"title":"MutS-mediated rapid and cost-effective error correction in in vitro DNA synthesis","authors":"Liting Jia , Shuang Wang , Dongmei Wang , Xingjiang Li , Jiong Hong","doi":"10.1016/j.nbt.2025.05.002","DOIUrl":"10.1016/j.nbt.2025.05.002","url":null,"abstract":"<div><div>The <em>in vitro</em> synthesis of DNA oligonucleotides and their subsequent assembly into longer target molecules represents a pivotal technique within the field of synthetic biology. However, the occurrence of side reactions and the inherent coupling efficiency of the synthesis process lead to the unavoidable introduction of errors into the resulting DNA. Consequently, there is a pressing demand for a straightforward, cost-effective, and efficient method for error correction. In this study, eleven Cbm3-Egfp-MutS fusion proteins were recombinantly expressed and purified, and their capacity to bind heteroduplex DNA was assessed. Among the MutS proteins, TaMutS and TtMutS exhibited thermal stability and effectively distinguished DNA containing mismatches. Following this, a simple, rapid, efficient, and economical error correction method was devised utilizing a homemade spin column composed of amorphous cellulose and a filter tip. The quantitative affinity of EcMutS, TaMutS, and TtMutS for all conceivable single-base errors was determined, and the efficacy of combining MutS proteins for error correction was evaluated. The error rate in synthesized DNA was reduced by a factor ranging from 2.15--8.17, with the material cost for a single reaction amounting to $0.032. The reaction volume was limited to 10 μL, and the reaction could be completed within 20 minutes.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"88 ","pages":"Pages 150-160"},"PeriodicalIF":4.5,"publicationDate":"2025-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144187442","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