Engineering in Life Sciences最新文献

{"title":"Evaluation of Yeast Alcohol Acetyltransferases for Ethyl Acetate Production in Clostridium ljungdahlii","authors":"Santiago T. Boto,&nbsp;Kareem Gerges,&nbsp;Bettina Bardl,&nbsp;Miriam A. Rosenbaum","doi":"10.1002/elsc.202400076","DOIUrl":"10.1002/elsc.202400076","url":null,"abstract":"<p>Sustainable chemical production from C₁ gaseous substrates, such as syngas or CO₂/H₂, can be achieved through gas fermentation. In gas fermentation, acetogenic bacteria are able to utilize oxidized inorganic carbon sources as the sole carbon source and electron acceptor, while reduced inorganic species are used as the electron donor. <i>Clostridium ljungdahlii</i>, a model acetogen, is only capable of reducing CO₂ to acetate and ethanol, with H₂ as electron donor. In order to expand the product profile of this bacterium, five alcohol acetyltransferases (AATs) from yeast were heterologously expressed in <i>C. ljungdahlii</i> to evaluate its potential to produce ethyl acetate. When growing on CO₂ and H₂, up to 7.38 ± 0.43 mg/L of ethyl acetate were produced. Using fructose as the main carbon and energy source, up to 23.15 ± 1.28 mg/L of ethyl acetate were produced. Ethanol and fumarate supplementation were able to boost ethyl acetate titers (up to 37.51 ± 9.44 mg/L). Hence, ethyl acetate production was enabled in <i>C. ljungdahlii</i> at low titers, which could be explained by the high energetic cost of operation of AATs, and their shown promiscuity. However, we also show that this opens the door to more complex esterification reactions of higher added value biomolecules.</p>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"25 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11756512/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143028214","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Polyporus umbellatus, A Precious Rare Fungus With Good Pharmaceutical and Food Value","authors":"Sizhu Ren,&nbsp;Hua Liu,&nbsp;Qing Sang,&nbsp;Yifan Sun,&nbsp;Liyan Li,&nbsp;Wenjie Chen","doi":"10.1002/elsc.202400048","DOIUrl":"10.1002/elsc.202400048","url":null,"abstract":"<p><i>Polyporus umbellatus</i> is a rare porous fungus that exhibits notable pharmacological activities. Particularly, due to its diuretic properties, it is considered an important source of targeted drugs for the treatment of kidney disease. Extensive research has been conducted on this fungus, focusing not only on its challenging cultivation techniques but also on its diverse array of medicinal ingredients, including polysaccharides and steroids. These active compounds demonstrate considerable variability and exhibit a wide range of medicinal properties. As a result, extracting, separating, and purifying these active compounds has become a subject of interest. This review aims to provide a comprehensive overview of the types, structures, and physicochemical properties of these active compounds. Additionally, the medicinal effects of <i>P. umbellatus</i> are thoroughly examined, offering valuable insights into the utilization of its resources and the rational development of medical fungi.</p>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"25 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-01-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11742960/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143002277","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Synthesis and Evaluation of PCL/Chitosan/CQD-Fe Magnetic Nanocomposite for Wound Healing: Emphasis on Gene Expression","authors":"Elham Maghareh Abed,&nbsp;Fatemeh Yazdian,&nbsp;Abbas Akhavan Sepahi,&nbsp;Behnam Rasekh","doi":"10.1002/elsc.202400038","DOIUrl":"10.1002/elsc.202400038","url":null,"abstract":"<p>The development of an effective and rapid method for healing the skin is of crucial importance. In this study, we prepared a porous scaffold made of polycaprolactone (PCL) and carbon quantum dots (CQDs), Fe, and Chitosan (Cs) as the scaffold core to cover the skin. Then evaluated antibacterial, biocompatibility, and wound healing properties as well as the expression of genes effective in wound healing. The PCL/Cs/CQD-Fe scaffold was synthesized via electrospinning and was evaluated of morphology, functional groups, and structure through Fourier transform infrared spectroscopy (FTIR), scanning electron microscope (SEM), and x-ray diffraction (XRD). The viability of the L929 fibroblast stem cells was obtained. The antibacterial effect, biocompatibility, and wound healing efficiency of the scaffold were investigated through minimum inhibitory concentration (MIC), (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), and tissue analysis. The relative expression of genes platelet-derived growth factor (PDGF), transforming growth factor beta (TGF-β), and matrix metalloproteinase-1 (MMP1) was assessed through RT-PCR. The results of SEM showed the successful integration of the PCL scaffold with CQD-Fe and Cs. The mean size of PCL/Cs/CQD-Fe nanocomposite was in the range of 0.135–32.6 nm. The results of FTIR showed the formation of a link between CQD nanoparticles and Fe. The vibrating-sample magnetometer (VSM) proved the super para magnetism of the CQD-Fe magnetic nanoparticles (0.38 emu/g). The MIC of Cs/CQD-Fe against <i>Staphylococcus aureus</i> and <i>Escherichia coli</i> bacteria was 0.08 and 0.04 µg/mL, respectively. The mean expression of genes TGF-β and PDGF in the nanocomposite group were 0.05 and 0.015 on day 5 and 0.18 and 0.34 on day 15 and significantly increased after 15 days, whereas the mean expression of MMP1 in the nanocomposite group was 0.63 on day 5 and 0.12 on day 15 and significantly decreased after 15 days. According to the histological analysis, the thickest layer on Day 15 pertained to the nanocomposite group. Our findings indicated that PCL/Cs/CQD-Fe can improve skin regeneration due to its antibacterial effect, biocompatibility, and non-toxicity. This biocompatible nanocomposite is a scaffold that can be used for covering the skin.</p>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"25 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-01-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11742958/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143002279","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Optimizations of Placenta Extracellular Matrix-Loaded Silk Fibroin/Alginate 3D-Printed Scaffolds Structurally and Functionally for Bone Tissue Engineering","authors":"Zahra Bashiri,&nbsp;Zahra Khosrowpour,&nbsp;Ali Moghaddaszadeh,&nbsp;Davod Jafari,&nbsp;Sanaz Alizadeh,&nbsp;Hajar Nasiri,&nbsp;Houman Parsaei,&nbsp;Zahra Keshtkaran,&nbsp;Meghdad Abdollahpour-Alitappeh,&nbsp;Farshad Bargrizaneh,&nbsp;Behzad Rezaei,&nbsp;Sara Simorgh,&nbsp;Mazaher Gholipourmalekabadi","doi":"10.1002/elsc.202400085","DOIUrl":"10.1002/elsc.202400085","url":null,"abstract":"<p>Recent interest has been focused on extracellular matrix (ECM)–based scaffolds totreat critical-sized bone injuries. In this study, urea was used to decellularize and solubilize human placenta tissue. Then, different concentrations of ECM were composited with 8% alginate (Alg) and 12% silk fibroin (SF) for printing in order to produce a natural 3D construct that resembled bone tissue. The physical and biological features of the printed structures were evaluated entirely in vitro. Finally, a rat model was employed to examine the optimal 3D printed scaffold (5% ECM) as a bone transplant for the healing of cranial bone lesions. The present investigation demonstrated that decellularizing placental tissue fragments led to efficient removal of cell debris. In addition, a remarkable improvement in the printed scaffolds' mechanical and biological properties was observed by increasing the ECM concentration. The histology studies and real-time PCR results demonstrated the acceleration of bone regeneration in the bone lesions treated with 5%ECM-SF/Alg at 4 and 8 weeks after implantation. Overall, these results proved that the placental ECM-printed scaffolds could potentially construct biomimetic grafts to reconstruct significant bone defects and now promise to proceed with clinical studies.</p>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"25 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11717148/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142970023","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Reaction Engineering for Asymmetric R-/S-PAC Synthesis With Ephedrine or Pseudoephedrine Dehydrogenase in Pickering Emulsion","authors":"Reynaldo Jr. Carubio,&nbsp;Bao-Hsiang Wang,&nbsp;Marion B. Ansorge-Schumacher","doi":"10.1002/elsc.202400069","DOIUrl":"https://doi.org/10.1002/elsc.202400069","url":null,"abstract":"<p>The synthesis of enantiopure α-hydroxy ketones, particularly <i>R</i>- and <i>S</i>-phenylacetylcarbinol (PAC), represents an important process in the pharmaceutical industry, serving as a pivotal step in the production of drugs. Recently, two novel enzymes, ephedrine dehydrogenase (EDH) and pseudoephedrine dehydrogenase (PseDH), have been described. These enzymes enable the specific reduction of 1-phenyl-1,2-propanedione (PPD) to <i>R</i>-PAC and <i>S</i>-PAC, respectively. In this study, we transferred these enzymes into Pickering emulsions, which is an attractive reaction set-up for large-scale synthesis. The bioactive w/o Pickering emulsion (bioactive Pickering emulsion [BioPE]), in which methyl <i>tert</i>-butyl ether served as the continuous phase, was stabilized by silica nanoparticles. Formate dehydrogenase from <i>Rhodococcus jostii</i> was utilized for cofactor regeneration. Given the considerable complexity of the BioPE, this reaction system underwent a first-time application of design of experiment (DOE) for systematic engineering. A definitive screening design was employed to identify significant factors affecting space-time yield (STY) and conversion. Response surface methodology was used to optimize the conditions, resulting in the observation of a high STY of 4.2 g L⁻¹ h⁻¹ and a conversion of 83.2% for BioPE with EDH, and an STY of 4.4 g L⁻¹ h⁻¹ and a conversion of 64.5% for BioPE with PseDH.</p>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"25 2","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-01-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.202400069","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143455934","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Investigating Ultrafiltration Membranes and Operation Modes for Improved Lentiviral Vector Processing","authors":"Jennifer J. Labisch,&nbsp;Maria Evangelopoulou,&nbsp;Tobias Schleuß,&nbsp;Andreas Pickl","doi":"10.1002/elsc.202400057","DOIUrl":"10.1002/elsc.202400057","url":null,"abstract":"<p>The demand for lentiviral vectors (LVs) as tools for ex vivo gene therapies is ever-increasing. Despite their promising applications, challenges in LV production remain largely due to the fragile envelope, which challenges the maintenance of vector stability. Thus, downstream processing optimization to enhance efficiency, yield, and product quality is necessary. This study investigated the influence of membrane types and filtration devices during ultrafiltration (UF). Nine different membrane materials consisting of polyethersulfone (PES), regenerated cellulose, or Hydrosart, with distinct molecular weight cutoffs, were evaluated in stirred cells, centrifugal ultrafilters, and crossflow cassettes. The evaluation was based on the ability to retain infectious LV particles and remove impurities. The analysis revealed that a reinforced 100 kDa PES and a 300 kDa Hydrosart membrane had the best overall ability to concentrate infectious LVs and remove DNA, especially when operated in a stirred cell. Challenges were seen in the nonoptimized crossflow cassette process, where infectious LV recovery was generally lower compared to other devices. We demonstrated that membrane material and filtration device have a direct impact on the efficiency of LV UF.</p>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"25 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-01-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11717145/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142970019","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Consecutive Genome Engineering Method Reveals a New Phenotype and Regulation of Glucose and Glycerol Utilization in Clostridium Pasteurianum","authors":"Tom Nguyen,&nbsp;Luca W. G. Meleski,&nbsp;Minu P. Belavatta,&nbsp;Sivasubramanian Gurumoorthi,&nbsp;Chijian Zhang,&nbsp;Anna-Lena Heins,&nbsp;An-Ping Zeng","doi":"10.1002/elsc.202400026","DOIUrl":"10.1002/elsc.202400026","url":null,"abstract":"<p><i>Clostridium pasteurianum</i> is a microorganism for production of 1,3-propanediol (1,3-PDO) and butanol, but suffers from lacking genetic tools for metabolic engineering to improve product titers. Furthermore, previous studies of <i>C. pasteurianum</i> have mainly focused on single genomic modification. The aim of this work is the development and application of a method for modification of multiple gene targets in the genome of <i>C. pasteurianum</i>. To this end, a new approach for consecutive genome engineering is presented for the first time using a method based on endogenous CRISPR-Cas machineries. A total of three genome modifications were consecutively introduced in the same mutant and the effect of combined changes on the genome was observed by 39% decreased specific glycerol consumption rate and 29% increased 1,3-PDO yield in mixed substrate fermentations at laboratory scale in comparison to the wildtype strain. Additionally, examination of the phenotype of the generated mutant strain led to discovery of 2,3-butanediol (2,3-BDO) production of up to 0.48 g L⁻¹, and this metabolite was not reported to be produced by <i>C. pasteurianum</i> before. The developed procedure expands the genetic toolkit for <i>C. pasteurianum</i> and provides researchers an additional method which contributes to improved genetic accessibility of this strain.</p>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"25 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2025-01-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11717147/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142969969","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cover Picture: Engineering in Life Sciences 12'24","authors":"","doi":"10.1002/elsc.202470121","DOIUrl":"https://doi.org/10.1002/elsc.202470121","url":null,"abstract":"","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"24 12","pages":""},"PeriodicalIF":3.9,"publicationDate":"2024-12-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.202470121","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142867991","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Rheology and Culture Reproducibility of Filamentous Microorganisms: Impact of Flow Behavior and Oxygen Transfer During Salt-Enhanced Cultivation of the Actinomycete Actinomadura namibiensis","authors":"René Hanke,&nbsp;Jonas Lohr,&nbsp;Leon Poduschnick,&nbsp;Sebastian Tesche,&nbsp;Luc Fillaudeau,&nbsp;Jochen Büchs,&nbsp;Rainer Krull","doi":"10.1002/elsc.202400078","DOIUrl":"https://doi.org/10.1002/elsc.202400078","url":null,"abstract":"<p>Analyzing the relationship between cell morphology, rheological characteristics, and production dynamics of cultivations with filamentous microorganisms is a challenging task. The complex interdependencies and the commonly low reproducibility of heterogeneous cultivations hinder the bioprocess development of commercially relevant production systems. The present study aims to characterize process parameters in <i>Actinomadura namibiensis</i> shake flask cultures to gain insights into relationships between culture behavior and rheological characteristics during salt-enhanced labyrinthopeptin A1 production. Plate–plate (PP) and vane–cup rheometer measurements of viscous model fluids and culture broths are compared, revealing a more uniform distribution of broth when measured with the PP system. Additionally, rheological characteristics and culture performance of <i>A. namibiensis</i> cultures are evaluated using online data of the specific power input and the oxygen transfer rate. It is demonstrated that salt-enhancement labyrinthopeptin A1 production by the addition of 50 mM (NH₄)₂SO₄ increases the apparent viscosity of the <i>A. namibiensis</i> culture by four-fold and significantly reduces the reproducibility of the culture resulting in a 46 h difference in lag-phase duration. This approach demonstrates that the culture behavior of complex filamentous cell morphologies is challenging to decipher, but online monitoring of rheology and oxygen transfer can provide valuable insights into the cultivation dynamics of filamentous microbial cultures.</p>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"25 2","pages":""},"PeriodicalIF":3.9,"publicationDate":"2024-11-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.202400078","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143456079","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Succinic Acid Production With Actinobacillus succinogenes –Influence of an Electric Potential on the Intercellular NADH/NAD+ Balance","authors":"Jan-Niklas Hengsbach,&nbsp;Marcel Cwienczek,&nbsp;Wolfgang Laudensack,&nbsp;Judith Stiefelmaier,&nbsp;Nils Tippkötter,&nbsp;Roland Ulber","doi":"10.1002/elsc.202400053","DOIUrl":"10.1002/elsc.202400053","url":null,"abstract":"<p>Bioelectrochemical systems (BESs) offer a sustainable method for chemical production, including the enhanced production of succinic acid. By combining fermentation with BES, it could be possible to achieve sustainable succinic acid production and CO₂ fixation using <i>Actinobacillus succinogenes</i>. In literature, the potential application of BES is commonly associated with increased succinate yields, as it is expected to enhance the availability of NADH, thereby influencing the intracellular nicotinamide adenine dinucleotide (NADH/NAD⁺) balance. However, it remains unclear whether BES can improve NADH regeneration and achieve higher NADH/NAD⁺ ratios across all growth phases of <i>A. succinogenes</i>. This study investigates the impact of an applied electrical potential on the intracellular NADH/NAD⁺ ratio during an electrochemical-assisted fermentation process. Using an adapted high-performance liquid chromatography method with a Supelcosil LC-18-T column, it was demonstrated that NADH availability in BES, particularly during the stationary growth phase, improved by up to 1.98-fold compared to the control. This enhancement in reducing power led to a succinate yield of 0.747 ± 0.01 g g⁻¹, representing a 15.65% increase compared to a fermentation without electrochemical assistance. These findings support the expectation that the use of BES could enhance the competitiveness of bio-based succinate production.</p>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"25 1","pages":""},"PeriodicalIF":3.9,"publicationDate":"2024-11-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11717146/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142970025","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}