New biotechnology最新文献_第9页

Future food court in an urban shopping center can supply feedstocks for microbial protein production 未来城市购物中心的美食广场可以为微生物蛋白的生产提供原料。

IF 4.9 2区生物学

New biotechnology Pub Date : 2025-11-25 Epub Date: 2025-07-23 DOI: 10.1016/j.nbt.2025.07.007

Terhi Viinikanoja , Eevi Haajanen , Maria Pajumo, Amanda Lillberg, Emilia Nordlund, Anneli Ritala

{"title":"Future food court in an urban shopping center can supply feedstocks for microbial protein production","authors":"Terhi Viinikanoja , Eevi Haajanen , Maria Pajumo, Amanda Lillberg, Emilia Nordlund, Anneli Ritala","doi":"10.1016/j.nbt.2025.07.007","DOIUrl":"10.1016/j.nbt.2025.07.007","url":null,"abstract":"<div><div>Food courts in urban shopping centers can be future sources of feedstock for microbial protein production. This study explored whether filamentous fungi <em>Trichoderma reesei</em>, <em>Paecilomyces variotii</em> and <em>Rhizopus oligosporus</em> can be cultivated on growth media prepared from orange peels, soft drink waste-mix, expired bread or spent coffee grounds, all existing and potential by-products of future food courts. Furthermore, artificial urine to describe human urine as a future nitrogen source was investigated. Side stream pretreatments facilitated sugar releases up to 171 g L<sup>−1</sup> (expired bread). Cultivation media were prepared from pretreated side streams with 40 g L<sup>−1</sup> sugar. Flask cultivations supplemented with nitrogen and micronutrient increased fungal biomass production. From the nitrogen sources tested, yeast extract (10 g L<sup>−1</sup>) proved effective in media based on soft drink waste-mix and orange peel juice. For the latter artificial urine (5 g L<sup>−1</sup> urea) worked as well. The effects of these nitrogen supplements, along with pH, on biomass production were further optimized using design of experiment methodology and validated in lab-scale bioreactor cultivations. The bioreactor cultivations of <em>P. variotii</em> and <em>T. reesei</em> with soft drink waste-mix or orange peel juice supplemented with yeast extract (13 – 15 g L<sup>−1</sup>) resulted in good biomass production up to 37.8 g L<sup>−1</sup> and 15 g L<sup>−1</sup> dry weight with total protein contents reaching 25 % and 35 % for <em>P. variotii</em> and <em>T. reesei</em>, respectively. Amino acid analyses revealed balanced essential amino acid distributions for human consumption, confirming that future food court by-products are viable substrates for sustainable fungal protein production in a circular economy.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"89 ","pages":"Pages 152-162"},"PeriodicalIF":4.9,"publicationDate":"2025-11-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144718159","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

tiRNA: An efficient and controllable gene silencing technology via translation inhibition. tiRNA：一种有效、可控的翻译抑制基因沉默技术。

IF 4.9 2区生物学

New biotechnology Pub Date : 2025-11-25 Epub Date: 2025-08-06 DOI: 10.1016/j.nbt.2025.07.010

Bei Xia, Jiajing Cai, Zhilin He, Qubo Zhu

{"title":"tiRNA: An efficient and controllable gene silencing technology via translation inhibition.","authors":"Bei Xia, Jiajing Cai, Zhilin He, Qubo Zhu","doi":"10.1016/j.nbt.2025.07.010","DOIUrl":"10.1016/j.nbt.2025.07.010","url":null,"abstract":"<p><p>RNA-targeted therapies have emerged as a revolutionary breakthrough in biomedicine recently, offering unprecedented precision in regulating gene expression. Among these, steric blocking oligonucleotides (SBOs) represent a unique class of therapeutics that function through a steric blocking mechanism, allowing for reversible effects without RNA degradation. However, designing SBOs is challenging due to the lack of universal guidelines and the need for detailed analysis of mRNA/pre-mRNA characteristics, key site distributions, and RNA-binding protein interactions. To address this, we developed a novel aptamer-based technology called translation inhibition RNA (tiRNA), which inhibits mRNA translation by linking an eIF4G-targeting aptamer to a reverse complementary sequence of the target gene's 5'-UTR. This approach leverages the typical translation initiation mechanism of mRNA and employs a straightforward design strategy to specifically target and inhibit the translation of selected mRNAs without affecting others or causing RNA degradation. The efficacy of tiRNA is comparable to that of siRNA, providing precision, safety, and controllability for treating diseases linked to protein overexpression. Moreover, the effects of tiRNA can be reversed using a specially designed neutralizing strand, restoring normal mRNA translation and enhancing treatment controllability and personalization. This method holds significant potential for applications in cancer, gene therapy, and other fields requiring precise regulation of protein expression.</p>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":" ","pages":"177-190"},"PeriodicalIF":4.9,"publicationDate":"2025-11-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144799697","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 : 2025-11-25 Epub 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":"2025-11-25","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

Creation of colorless transparent tilapia using CRISPR/Cas9 mediated multi-gene mutation 利用CRISPR/Cas9介导的多基因突变培育无色透明罗非鱼。

IF 4.9 2区生物学

New biotechnology Pub Date : 2025-11-25 Epub Date: 2025-07-31 DOI: 10.1016/j.nbt.2025.07.009

Guangyuan Liang , Baoyue Lu , Shengfei Dai, Minghui Li, Jiawen Yao, Hao Liu, Xiayue Liu, Xingyong Liu, Deshou Wang

{"title":"Creation of colorless transparent tilapia using CRISPR/Cas9 mediated multi-gene mutation","authors":"Guangyuan Liang , Baoyue Lu , Shengfei Dai, Minghui Li, Jiawen Yao, Hao Liu, Xiayue Liu, Xingyong Liu, Deshou Wang","doi":"10.1016/j.nbt.2025.07.009","DOIUrl":"10.1016/j.nbt.2025.07.009","url":null,"abstract":"<div><div>Transparent mutant fish have been obtained from small-sized fish (medaka, zebrafish, and killifish), and are often used as experimental models in biological and medical research. However, transparent fish have never been created in medium- or large-sized fish through gene editing. In this study, mutants without xanthophores, erythrophores and pigmented melanophores were first obtained by crossing to form <em>tyrb;csf1ra</em> double mutant tilapia. Subsequently, single mutants lacking reflective platelets without visible iridophores were obtained by mutation of <em>pnp4a</em> and <em>tfec</em> using CRISPR/Cas9 gene editing, the recovery of iridophores were observed in <em>pnp4a</em> mutants but not in <em>tfec</em> mutants in the later stage. In addition, we also found that <em>tfec</em> mutation led to a decrease in number of melanophores and diameter of melanophores/erythrophores. Finally, transparent triple mutant homozygotes without visible pigment cells were obtained by triple mutation of <em>tyrb</em>;<em>csf1ra</em>;<em>pnp4a</em> (named <em>amber</em>) and <em>tyrb</em>;<em>csf1ra</em>;<em>tfec</em> (named <em>ruby</em>), through crossing of double and single mutants. The two mutant lines were no longer transparent at 60 dpf (days post fertilization) and 120 dpf, respectively, due to the recovery of iridophores in the former, and thickening of the body wall in the latter. In summary, this study created two transparent lines without visible pigment cells through aggregation of multiple gene mutation and crossing. The transparent fish is suitable for in vivo imaging, and is currently the only medium-sized transparent fish obtained through gene editing.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"89 ","pages":"Pages 163-176"},"PeriodicalIF":4.9,"publicationDate":"2025-11-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144768811","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 : 2025-11-25 Epub 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":"2025-11-25","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

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 : 2025-11-25 Epub 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":"2025-11-25","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

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 : 2025-11-25 Epub 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":"2025-11-25","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

Enhancing 2,3-butanediol and acetoin production from brewer's spent grain hemicellulosic hydrolysate through bacterial co-cultivation 通过细菌共培养提高啤酒废谷物半纤维素水解产物2,3-丁二醇和乙糖的产量

IF 4.5 2区生物学

New biotechnology Pub Date : 2025-09-25 Epub Date: 2025-03-24 DOI: 10.1016/j.nbt.2025.03.006

Juan C. López-Linares , Erlinda Rama , María Teresa García-Cubero , Mónica Coca , Caroline L. Perez , Celina K. Yamakawa , Giuliano Dragone , Solange I. Mussatto

{"title":"Enhancing 2,3-butanediol and acetoin production from brewer's spent grain hemicellulosic hydrolysate through bacterial co-cultivation","authors":"Juan C. López-Linares , Erlinda Rama , María Teresa García-Cubero , Mónica Coca , Caroline L. Perez , Celina K. Yamakawa , Giuliano Dragone , Solange I. Mussatto","doi":"10.1016/j.nbt.2025.03.006","DOIUrl":"10.1016/j.nbt.2025.03.006","url":null,"abstract":"<div><div>This study evaluated bacterial co-cultivation as a strategy to mitigate brewer’s spent grain (BSG) hemicellulosic hydrolysate toxicity, aiming to enhance 2,3-butanediol (2,3-BDO) and acetoin production through fermentation. Co-culture of <em>Paenibacillus polymyxa</em> with <em>Pseudomonas alloputida</em> or <em>Rhodococcus</em> sp. was assessed using synthetic medium and BSG hydrolysate. Attention was given to removing inhibitory compounds, including lignin-derived phenolics, hydroxymethylfurfural, furfural, and acetic acid, through microbial detoxification during co-cultivation. Various fermentation temperatures (30, 34, and 37 °C) and initial cell concentrations (OD<sub>600</sub> of 0.05 and 0.1) were tested. Both <em>P. polymyxa</em> and <em>Rhodococcus</em> sp. effectively removed inhibitory compounds present in the medium. Co-cultures with <em>Rhodococcus</em> sp. exhibited higher sugar consumption rates (1.01 vs 0.88 g/L·h) than <em>P. polymyxa</em> monoculture, efficiently utilizing glucose, xylose, and arabinose, producing 2,3-BDO and acetoin. In co-culture with <em>Rhodococcus</em> sp., concentration (3.7 g/L), yield (0.14 g/g) and productivity (0.10 g/L·h) of 2,3-BDO at 34 °C considerably surpassed that of the <em>P. polymyxa</em> monoculture, with an increase of up to 48 %. These findings highlight the potential of co-cultures, especially with <em>Rhodococcus</em> sp., to alleviate inhibitory compound impacts when using complex media for fermentation. This study represents the first exploration of 2,3-BDO and acetoin production from BSG hemicellulosic hydrolysates using co-cultures.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"88 ","pages":"Pages 22-31"},"PeriodicalIF":4.5,"publicationDate":"2025-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143705413","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

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 : 2025-09-25 Epub 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":"2025-09-25","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

Screening assay for polyester hydrolyzing microorganisms using fluorescence-labeled poly(butylene adipate) 利用荧光标记聚己二酸丁二醇酯筛选聚酯水解微生物的检测方法。

IF 4.5 2区生物学

New biotechnology Pub Date : 2025-09-25 Epub Date: 2025-04-05 DOI: 10.1016/j.nbt.2025.03.007

Bettina Semler , Karin Binder , Doris Ribitsch , Alessandro Pellis , Georg M. Guebitz

{"title":"Screening assay for polyester hydrolyzing microorganisms using fluorescence-labeled poly(butylene adipate)","authors":"Bettina Semler , Karin Binder , Doris Ribitsch , Alessandro Pellis , Georg M. Guebitz","doi":"10.1016/j.nbt.2025.03.007","DOIUrl":"10.1016/j.nbt.2025.03.007","url":null,"abstract":"<div><div>Despite recent advances, there is still a demand for more efficient enzymes hydrolyzing synthetic polymers. Automated high throughput screening strategies of microorganisms from different environments could yield novel enzymes but require specific methods for detection of polymer hydrolysis in complex matrices. Here, 5-carboxy-fluorescein (5-FAM) was covalently coupled to poly(butylene adipate) (PBA) and blended at 1 %, 5 % and 10 % w/w concentrations with non-labeled PBA. Hydrolysis of PBA by the Thc_Cut1 cutinase from <em>Thermobifida cellulosilytica</em> was confirmed via quantification of the released monomers 1,4-butanediol and adipic acid, weight loss and FTIR analysis. Upon incubation with Thc_Cut1, hydrolysis of all three fluorescent labeled PBA blends lead to a clear fluorescence increase of up to 4000 RFU while no signal change was detected for the blank and for heat-inactivated enzyme (signal below 500 RFU). In a next step, as a model organism <em>Pichia pastoris</em> expressing the identical cutinase was cultivated in the presences of labeled PBA. Despite the complex matrix, a fluorescence increase of up to 500 RFU was observed for <em>P. pastoris</em> expressing the enzyme while no significant signal change was seen for the control strain (lacking Thc_Cut1 expression). Likewise, extracellular enzymes from the fungi <em>Fusarium solani</em> and <em>Alternaria alternata</em> hydrolyzed labeled PBA leading to fluorescence increases of 1328 and 1187 RFU. This indicates that 5-FAM covalently coupled to polymers could be used for development of simple and high throughput screening platforms to identify polymer decomposing microorganisms and enzymes.</div></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":"88 ","pages":"Pages 39-45"},"PeriodicalIF":4.5,"publicationDate":"2025-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143803799","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}

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