Biotechnology and Bioengineering最新文献_第2页

A Bioprocess Engineering Approach to Boost Selection of Fully Segregated Transformants in Cyanobacteria. 促进蓝藻中完全分离转化体选择的生物过程工程方法。

IF 3.8 2区生物学

Biotechnology and Bioengineering Pub Date : 2025-07-14 DOI: 10.1002/bit.70024

Cecilia Salvagnini,Eliana Gasparotto,Veronica Lucato,Elisabetta Bergantino,Matteo Ballottari,Elena Barbera,Nico Betterle,Eleonora Sforza

{"title":"A Bioprocess Engineering Approach to Boost Selection of Fully Segregated Transformants in Cyanobacteria.","authors":"Cecilia Salvagnini,Eliana Gasparotto,Veronica Lucato,Elisabetta Bergantino,Matteo Ballottari,Elena Barbera,Nico Betterle,Eleonora Sforza","doi":"10.1002/bit.70024","DOIUrl":"https://doi.org/10.1002/bit.70024","url":null,"abstract":"Cyanobacteria are photoautotrophic microorganisms with significant applications in biotechnology. Although many cyanobacteria, including Picosynechococcus sp. (formerly called Synechococcus sp.) PCC 11901 (Picosynechococcus) and Synechocystis sp. PCC 6803 (Synechocystis), are readily and naturally transformable, their polyploidy poses a major challenge. To obtain a stable phenotype, transgenic strains must be fully segregated, i.e. mutations must appear in all chromosome copies. Traditional protocols rely on re-streaking of colonies on increasingly selective plates, a time-intensive laboratory procedure that requires continuous intervention from the operator. This study proposes an alternative protocol that combines transformation in a batch system in liquid culture with transformant selection in a continuous-flow stirred-tank reactor system. This protocol led to the successful selection of homoplasmic transformants of Picosynechococcus containing, alternatively, an antibiotic resistance alone (construct \"SmR\") or a more complex construct (\"bKT\") that leads to the accumulation of a ketocarotenoid. The stability of SmR transformants under semi-continuous cultivation in the absence of antibioticsf was tested for 42 days, proving their potential fitness to industrial cultivation conditions. The selection process was also validated on the model species Synechocystis, demonstrating its applicability to other cyanobacterial strains.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"52 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-07-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144622025","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

Regulating Transport Preferences of Fucosylated Sugars: Revealing Transport Mechanisms via Sugar Efflux Transporter A Transformation Into a “Tight‐in, Tight‐out” Mode 调节聚焦糖的运输偏好：通过糖外排转运蛋白A转化为“紧进，紧出”模式揭示运输机制

IF 3.8 2区生物学

Biotechnology and Bioengineering Pub Date : 2025-07-11 DOI: 10.1002/bit.70019

Liu Wenxian, Sun Shengjie, Peng Jing, Zou Sini, Cheng Haina, Chen Zhu, Wang Yuguang, Zhou Hongbo

{"title":"Regulating Transport Preferences of Fucosylated Sugars: Revealing Transport Mechanisms via Sugar Efflux Transporter A Transformation Into a “Tight‐in, Tight‐out” Mode","authors":"Liu Wenxian, Sun Shengjie, Peng Jing, Zou Sini, Cheng Haina, Chen Zhu, Wang Yuguang, Zhou Hongbo","doi":"10.1002/bit.70019","DOIUrl":"https://doi.org/10.1002/bit.70019","url":null,"abstract":"Sugar transporters play a crucial role in cellular metabolism across diverse organisms, regulating essential biological processes through efficient substrate transport. Despite extensive research efforts, the structures and mechanisms of transporters responsible for sugars have remained elusive. In this study, we investigated the transport efficiency of the Escherichia coli sugar efflux transporter A (SetA) for lactose and fucosylated lactose. By employing site and combinatorial mutations, we obtained a mutant exhibiting approximately sixfold enhanced transporter efficiency for fucosylated lactose while retaining its potency for lactose transport. In this mutant, the fundamental amino acids responsible for recognizing the galactosyl moiety remained unchanged, yet the introduction of two face‐to‐face aromatic ring residues facilitated the enhanced recognition of the fucosyl moiety. This indicated the transformation of SetA from a universal transporter into a specific “tight‐in, tight‐out” transporter. Utilizing SetA‐based structural modeling, we mapped and investigated mutations associated with diseases. The structural and biochemical insights from SET in this study offer a valuable investigating framework for understanding substrate specificity mechanisms of fucosylated sugar transporters and, by extension, other transporters in broader contexts.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"22 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144603105","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 Setup for Automatic Raman Measurements in High‐Throughput Experimentation 高通量实验中的自动拉曼测量装置

IF 3.8 2区生物学

Biotechnology and Bioengineering Pub Date : 2025-07-11 DOI: 10.1002/bit.70006

Christoph Lange, Simon Seidel, Madeline Altmann, Daniel Stors, Annina Kemmer, Linda Cai, Stefan Born, Peter Neubauer, M. Nicolas Cruz Bournazou

{"title":"A Setup for Automatic Raman Measurements in High‐Throughput Experimentation","authors":"Christoph Lange, Simon Seidel, Madeline Altmann, Daniel Stors, Annina Kemmer, Linda Cai, Stefan Born, Peter Neubauer, M. Nicolas Cruz Bournazou","doi":"10.1002/bit.70006","DOIUrl":"https://doi.org/10.1002/bit.70006","url":null,"abstract":"High‐throughput (HT) experimentation is transforming biotechnology by enabling systematic exploration of complex multi‐dimensional experimental conditions. However, current analytical methods are often unable to handle the rapid pace of sample generation in HT workflows. This study presents an integrated system of physical devices and software to automate and accelerate Raman spectral measurements in HT‐facilities. The setup simultaneously handles eight parallel L samples delivered by a pipetting robot, completing measurement, handling, cleaning, and concentration prediction within 45 s per sample. We introduce a machine learning model to predict metabolite concentrations from Raman spectra, achieving mean absolute errors of for glucose and for acetate during Escherichia coli cultivations. This approach enables consistent high‐throughput spectral data collection for fermentation monitoring, calibration, and offline analysis, supporting the generation of extensive datasets, enabling the training of more robust and generalizable machine learning models.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"11 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144603104","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

Engineered Coenzyme A Biosynthesis and Butyrate Transporter Drives High‐Efficient Butyrate Synthesis in Escherichia coli 工程辅酶A生物合成和丁酸转运蛋白驱动大肠杆菌高效合成丁酸盐

IF 3.8 2区生物学

Biotechnology and Bioengineering Pub Date : 2025-07-11 DOI: 10.1002/bit.70018

Jinhui Li, Yi Xing, Xin Wang, Tong Zhu, Feiyu Fan, Hongtao Xu, Peipei Han, Jun Cai, Xinna Zhu, Xueli Zhang

{"title":"Engineered Coenzyme A Biosynthesis and Butyrate Transporter Drives High‐Efficient Butyrate Synthesis in Escherichia coli","authors":"Jinhui Li, Yi Xing, Xin Wang, Tong Zhu, Feiyu Fan, Hongtao Xu, Peipei Han, Jun Cai, Xinna Zhu, Xueli Zhang","doi":"10.1002/bit.70018","DOIUrl":"https://doi.org/10.1002/bit.70018","url":null,"abstract":"Butyric acid is a four‐carbon fatty acid with a range of applications in chemical, food and pharmaceutical industries. In this study, a heterologous butyrate biosynthetic pathway was engineered in Escherichia coli, which was afflicted by low titer and low yield. To address these issues, two key strategies for metabolic engineering of E. coli were implemented by enhancing coenzyme A (CoA) biosynthesis and optimizing butyrate transport. First, the CoA biosynthesis pathway was engineered through alleviating CoA‐mediated inhibition, and enhancing the supply of pantothenate and cysteine precursors. Second, a TolC‐associated MdtEF efflux pumps was identified and optimized to mitigate butyrate reuptake. The combined implementation in strain JH016 led to 11.1‐fold and 86% increase of butyrate titer and yield, resulting in production of 21.12 g/L butyrate with a yield of 0.95 mol/mol. Our results suggested that CoA engineering and butyrate transporter optimization had a synergistic effect on butyrate production. Furthermore, these strategies could be broadly utilized for the production of various other useful chemicals in the fields of metabolic engineering and synthetic biology.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"71 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144603349","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

Inside Front Cover Image, Volume 122, Number 8, August 2025 封面内图，第122卷，第8期，2025年8月

IF 3.5 2区生物学

Biotechnology and Bioengineering Pub Date : 2025-07-07 DOI: 10.1002/bit.70023

Elnaz Khorasani, Bahman Vahidi

引用次数: 0

Biotechnology and Bioengineering: Volume 122, Number 8, August 2025 生物技术和生物工程：第122卷，第8号，2025年8月

IF 3.5 2区生物学

Biotechnology and Bioengineering Pub Date : 2025-07-07 DOI: 10.1002/bit.28748

引用次数: 0

Front Cover Image, Volume 122, Number 8, August 2025 封面图片，第122卷，第8期，2025年8月

IF 3.5 2区生物学

Biotechnology and Bioengineering Pub Date : 2025-07-07 DOI: 10.1002/bit.70022

Anton Lindig, Georg Hubmann, Stephan Lütz

引用次数: 0

Sulfate Reduction in the Hydrogen‐Based Membrane Biofilm Reactor Receiving Calcium Reduced Phosphogypsum Water 接受钙还原磷石膏水的氢基膜生物膜反应器中的硫酸盐还原

IF 3.8 2区生物学

Biotechnology and Bioengineering Pub Date : 2025-07-05 DOI: 10.1002/bit.70015

Anwar Alsanea, Ayoub Bounaga, Karim Lyamlouli, Youssef Zeroual, Rachid Boulif, Chen Zhou, Bruce Rittmann

{"title":"Sulfate Reduction in the Hydrogen‐Based Membrane Biofilm Reactor Receiving Calcium Reduced Phosphogypsum Water","authors":"Anwar Alsanea, Ayoub Bounaga, Karim Lyamlouli, Youssef Zeroual, Rachid Boulif, Chen Zhou, Bruce Rittmann","doi":"10.1002/bit.70015","DOIUrl":"https://doi.org/10.1002/bit.70015","url":null,"abstract":"Phosphogypsum (PG), a byproduct of phosphate mining, contains sulfate that can be leached and converted to elemental sulfur, thus offering a sustainable opportunity to recover sulfur (S) as a step toward a circular economy. Calcium, at ~15 mM in PG leachate, creates inorganic precipitation that interferes with biological sulfate reduction, the first step of S recovery. Here, we evaluated the effectiveness of using cation‐exchange to lower the calcium concentration in water‐leached PG (PG water) delivered to a H2‐based membrane biofilm reactor (H2‐MBfR) employed to reduce sulfate to sulfide. A high sulfate flux (1 gS/m2‐day) and 65% sulfate reduction were achieved despite a high pH (10) resulting from base production during sulfate reduction. However, soluble sulfide was only 20% of the reduced S, possibly due to precipitation of sulfide, iron, and phosphate, and alkalinity analysis revealed possible formation of polysulfides. Shallow metagenomics of the biofilm documented that Desulfomicrobium was the dominant sulfate‐reducing bacterium, while Thauera, a mixotroph capable of sulfate reduction and sulfide oxidation, also was an important genus. The metagenomics also revealed the presence of methanogens and acetogens that competed for H2 and CO2. Although calcium removal from PG water improved sulfate reduction and reduced inorganic precipitation in the H2‐MBfR, soluble sulfide generation must be improved by supplying sufficient CO2 to moderate pH increase due to sulfate reduction and by controlling the H2‐delivery capacity to limit methanogens and acetogens.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"42 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-07-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144565957","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 Surface Plasmon Resonance‐Based Integrated Assay for Quantification and Glycosylation Characterization of Monoclonal Antibodies in Crude Heterogeneous Samples 基于表面等离子体共振的单克隆抗体定量和糖基化鉴定综合分析

IF 3.8 2区生物学

Biotechnology and Bioengineering Pub Date : 2025-07-05 DOI: 10.1002/bit.70016

Ilona Metayer, Catherine Forest‐Nault, Julie Guimond, Simon Joubert, Olivier Henry, Yves Durocher, Gregory De Crescenzo, Jimmy Gaudreault

{"title":"A Surface Plasmon Resonance‐Based Integrated Assay for Quantification and Glycosylation Characterization of Monoclonal Antibodies in Crude Heterogeneous Samples","authors":"Ilona Metayer, Catherine Forest‐Nault, Julie Guimond, Simon Joubert, Olivier Henry, Yves Durocher, Gregory De Crescenzo, Jimmy Gaudreault","doi":"10.1002/bit.70016","DOIUrl":"https://doi.org/10.1002/bit.70016","url":null,"abstract":"The rise in cancer, autoimmune, inflammatory, and infectious diseases in recent decades has led to a surge in the development of monoclonal antibodies (mAbs) therapies, now the most widely used family of biologics. To meet the growing global demand, biopharmaceutical industries are intensifying their production processes. One approach to achieve more efficient production of effective mAbs is to develop tools for real‐time quality monitoring. Specifically, the glycosylation profile of mAbs must be closely monitored, since it greatly impacts their therapeutic efficacy and innocuity, making it a critical quality attribute. In this study, we developed a surface plasmon resonance‐based integrated assay allowing for the simultaneous quantification and glycosylation characterization of mAbs in crude samples, hence permitting the at‐line analysis of bioreactor cell cultures. Thanks to the high specificity of the interaction between biosensor surface‐bound protein A and the Fc region of mAbs, we quantified crude IgG samples under mass transport limitations. Next, by flowing running buffer on the surface, impurities contained in the mAbs samples were washed away from the biosensor surface, allowing subsequent recording of the kinetics between the captured mAbs and injected FcγRII receptors. Of interest, with this strategy, we were able to quantify terminal galactosylation and core fucosylation of IgG lots, two important glycan modifications for mAb efficacy.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"48 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-07-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144566608","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

Transferrin Purification, Biophysical Characterization, and Lung Biodistribution in Sickle Cell Disease Mice. 镰状细胞病小鼠转铁蛋白纯化、生物物理特性和肺生物分布。

IF 3.8 2区生物学

Biotechnology and Bioengineering Pub Date : 2025-06-30 DOI: 10.1002/bit.70012

Shuwei Lu,Mohd A Khan,Saini Setua,Quintin O'Boyle,Kiruphagaran Thangaraju,Pedro Cabrales,Delaney C Swindle,David C Irwin,Paul W Buehler,Andre F Palmer

{"title":"Transferrin Purification, Biophysical Characterization, and Lung Biodistribution in Sickle Cell Disease Mice.","authors":"Shuwei Lu,Mohd A Khan,Saini Setua,Quintin O'Boyle,Kiruphagaran Thangaraju,Pedro Cabrales,Delaney C Swindle,David C Irwin,Paul W Buehler,Andre F Palmer","doi":"10.1002/bit.70012","DOIUrl":"https://doi.org/10.1002/bit.70012","url":null,"abstract":"Plasma transferrin (Tf) is the transport protein central to the process of iron recycling and metabolism. Holo-Tf serves as the body's pool of ferric iron, facilitating transport from tissues such as the intestine, liver, spleen, and finally bone marrow, where iron is incorporated into erythropoiesis. In sickle cell disease (SCD), iron overload is primarily caused by chronic blood transfusions in patients at risk of stroke or frequent acute pain crisis. However, we have identified that pulmonary vascular iron accumulation, independent of transfusion, is a driver of pulmonary hypertension in SCD patients and murine models. Therefore, we hypothesize that intra-pulmonary administration of apo-Tf localizes the protein to sites of iron accumulation within the lung, where reactive iron-driven pathology develops. This approach to therapeutic development focuses on optimizing administration using aerosol drug delivery, which can increase clinical compliance compared to subcutaneous or intravenous administration. The goal of this study was to purify apo-Tf using a novel process, perform biochemical characterization on the material, and test the proof of concept that apo-Tf protein can be delivered to lung regions where iron accumulation occurs in SCD pulmonary hypertension. We conclude that apo-Tf can be isolated from plasma Cohn fraction IV paste using a simple process and that characterization of the material identified a high-purity apo-Tf product with functional iron binding properties. Further, this material was administered to SCD mice to target pulmonary anatomical regions where pathology occurs. This data suggests an intriguing approach to iron chelation applicable to a relevant clinical population.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"47 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144521269","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