New biotechnology最新文献

{"title":"Virus inactivation using an electrically conducting virus filter in biopharmaceutical manufacturing process","authors":"","doi":"10.1016/j.nbt.2024.09.003","DOIUrl":"10.1016/j.nbt.2024.09.003","url":null,"abstract":"<div><p>Biopharmaceutical manufacturing processes using mammalian cells or plasma carry the risk of viral contamination. To mitigate these risks, it is essential to ensure viral clearance during the downstream process. Virus-retentive filters are used for size-based virus filtration, offering robust viral removal of more than 99.99%. However, virus breakthroughs have also been reported during virus filtration under certain conditions. In addition, these virus-retentive filters are disposable to ensure the safety of bioproducts, leading to significant costs and environmental concerns. In this study, innovative electrically conducting virus filters were fabricated using free-standing carbon veils (CV) and used to achieve additional virus inactivation after filtration. The viruses were captured in a CV-assisted virus filter, which was electrically heated using direct current to inactivate the viruses. This electrically conducting virus filter can inactivate viruses and can be reused up to five times. These results demonstrate that electrical conduction through electrical conducting damaged the phage capsid and eliminated the RNA genome, leading to bacteriophage inactivation. Moreover, it was confirmed that the electrically conducting virus filter could be reused up to five times without any changes to its physical or chemical structure. This study contributes to the reduction of process costs and environmental impacts by enabling the reuse of virus filters and enhancing the safety of the virus filtration process by preventing undesired virus breakthroughs.</p></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-09-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1871678424005442/pdfft?md5=c37609cc1abadedee77c02696abff6d2&pid=1-s2.0-S1871678424005442-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142227993","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}

{"title":"Hints from nature for a PHA circular economy: Carbon synthesis and sharing by Pseudomonas solani GK13","authors":"","doi":"10.1016/j.nbt.2024.09.002","DOIUrl":"10.1016/j.nbt.2024.09.002","url":null,"abstract":"<div><p>Polyhydroxyalkanoates (PHAs) are a well-known group of biodegradable and biocompatible bioplastics that are synthesised and stored by microorganisms as carbon and energy reservoirs. Extracellular PHA depolymerases (ePhaZs), secreted by a limited range of microorganisms, are the main hydrolytic enzymes responsible for their environmental degradation. <em>Pseudomonas</em> sp. GK13, initially identified as <em>P. fluorescens</em> GK13, produces PHA and a prototypic ePhaZ that specifically degrades mcl-PHA. In this study, a comprehensive characterization of strain GK13 was performed. The whole genomic sequence of GK13 was consolidated into one complete chromosome, leading to its reclassification as <em>P. solani</em> GK13. We conducted a detailed <em>in silico</em> examination of the bacteria genomic sequence, specifically targeting PHA metabolic functions. From the different growth conditions explored, PHA accumulation occurred only under carbon/nitrogen (C/N) imbalance, whereas ePhaZ production was induced even at balanced C/N ratios in mineral media. We extend our study to other bacteria belonging to the <em>Pseudomonas</em> genus revealing that the ePhaZ production capacity is closely associated with mcl-PHA synthesis capacity, as also suggested by metagenomic samples. This finding suggests that these types of microorganisms could contribute to the carbon economy of the microbial community, by storing PHA in carbon-rich times, and sharing it with the rest of the population during times of carbon scarcity through PHA hydrolysis. The conclusion pointed that carbon cycle metabolism performed by <em>P. solani</em> GK13 may contribute to the environmental circular economy at a microscopic scale.</p></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-09-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1871678424005430/pdfft?md5=7567945b424b16c69eddbbe82d513f30&pid=1-s2.0-S1871678424005430-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142154689","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}

{"title":"Characterization of the conversion system of natural rubber to poly(3-Hydroxyalkanoate) in Piscinibacter gummiphilus strain NS21T","authors":"","doi":"10.1016/j.nbt.2024.08.507","DOIUrl":"10.1016/j.nbt.2024.08.507","url":null,"abstract":"<div><p>Poly(3-hydroxyalkanoate) (PHA), a bacteria-synthesized biodegradable polyester, is a useful alternative to fossil resources, and current systems for its production rely predominantly on edible resources, raising concerns about microbial competition for nutrients. Therefore, we investigated mechanisms underlying PHA production from non-edible resources by <em>Piscinibacter gummiphilus</em> strain NS21^T. Strain NS21^T can utilize natural rubber as a carbon source on solid media and potentially produces PHA. Gas chromatography and nuclear magnetic resonance analyses of NS21^T cell extracts revealed the production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) and poly(3-hydroxybutyrate) from natural rubber and glucose, respectively. Transcriptional analysis suggested that <em>phaC</em> is involved in PHA production. An increased PHBV accumulation rate under nitrogen-limiting conditions indicates the potential of this strain to be used as a PHBV production enhancement strategy. Furthermore, the disruption of PHA depolymerase genes resulted in enhanced PHA production, indicating the involvement of these genes in PHA degradation. These findings highlight the potential of NS21^T for PHBV production from natural rubber, a non-edible resource.</p></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-08-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1871678424005417/pdfft?md5=d165ab1cdd6a7333ef1c4b76b4ffbdcd&pid=1-s2.0-S1871678424005417-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142110002","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}

{"title":"Phage display for discovery of anticancer antibodies","authors":"","doi":"10.1016/j.nbt.2024.08.506","DOIUrl":"10.1016/j.nbt.2024.08.506","url":null,"abstract":"<div><p>Antibodies and antibody-based immunotherapeutics are the mainstays of cancer immunotherapy. Expanding the repertoire of cancer-specific and cancer-associated epitopes targetable with antibodies represents an important area of research. Phage display is a powerful approach allowing the use of diverse antibody libraries to be screened for binding to a wide range of targets. In this review, we summarize the basics of phage display technology and highlight the advances in anticancer antibody identification and modification via phage display platform. Finally, we describe phage display-derived anticancer monoclonal antibodies that have been approved to date or are in clinical development.</p></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-08-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1871678424005405/pdfft?md5=d1ac89731a298ee2922b7863107f9137&pid=1-s2.0-S1871678424005405-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142073440","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}

{"title":"Polyethylene and related hydrocarbon polymers (“plastics”) are not biodegradable","authors":"","doi":"10.1016/j.nbt.2024.08.503","DOIUrl":"10.1016/j.nbt.2024.08.503","url":null,"abstract":"<div><p>Research on the biodegradation of polyethylene (PE), polystyrene (PS) and related polymers has become popular and the number of publications on this topic is rapidly increasing. However, there is no convincing evidence that the frequently claimed biodegradability of these so-called “plastics” really exists. Rather, a diffuse definition of the term “biodegradability” has led to the publication of reports showing either marginal weight losses of hydrocarbon polymers by the action of isolated bacterial strains or mechanical disintegration and polymer surface modification in case of hydrocarbon polymer-consuming insect larvae. Most of the data can be alternatively explained by the utilization of polymer impurities/additives, by the utilization of low molecular weight oligomers, and/or by physical fragmentation and subsequent loss of small fragments. Evidence for a (partial) biotic and/or abiotic oxidation of the amorphous polymer fraction and of surface-exposed hydrocarbon side chains is not sufficient to claim that PE is biodegradable. To the best of my knowledge, no report has been so far published in which substantial biodegradation and mineralization of PE or related (long chain length) hydrocarbon polymers to carbon dioxide has been convincingly demonstrated by the determination of the fate of carbon atoms in isotope-labeled polymers. It is disappointing that publications with a critical view on biodegradation of hydrocarbon polymers are not cited in most of these reports. The possibility should be considered that the rapidly expanding research field of hydrocarbon polymer biodegradation is chasing rainbows.</p></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-08-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1871678424005375/pdfft?md5=19fc1c6e925672401725a25f484ebcd0&pid=1-s2.0-S1871678424005375-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142056170","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}

{"title":"Cultivation optimization promotes ginsenoside and universal triterpenoid production by engineered yeast","authors":"","doi":"10.1016/j.nbt.2024.08.505","DOIUrl":"10.1016/j.nbt.2024.08.505","url":null,"abstract":"<div><p>Ginseng, a cornerstone of traditional herbal medicine in Asia, garnered significant attention for its therapeutic potential. Central to its pharmacological effects are ginsenosides, the primary active metabolites, many of which fall within the dammarane-type and share protopanaxadiol as a common precursor. Challenges in extracting protopanaxadiol and ginsenosides from ginseng arise due to their low concentrations in the roots. Emerging solutions involve leveraging microbial cell factories employing genetically engineered yeasts. Here, we optimized the fermentation conditions via the Design of Experiment, realizing 1.2 g/L protopanaxadiol in simple shake flask cultivations. Extrapolating the optimized setup to complex ginsenosides, like compound K, achieved 7.3-fold (0.22 g/L) titer improvements. Our adaptable fermentation conditions enable the production of high-value products, such as sustainable triterpenoids synthesis. Through synthetic biology, microbial engineering, and formulation studies, we pave the way for a scalable and sustainable production of bioactive compounds from ginseng.</p></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1871678424005387/pdfft?md5=c2ae2ff1565b1d5e027a266007c6dba2&pid=1-s2.0-S1871678424005387-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142056167","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}

{"title":"PhagoScreener: A novel phagogram platform based on a capillary-wave microbioreactor","authors":"","doi":"10.1016/j.nbt.2024.08.502","DOIUrl":"10.1016/j.nbt.2024.08.502","url":null,"abstract":"<div><p>Due to the overuse of antibiotics, the number of multidrug-resistant pathogen bacteria is rising in recent years posing a serious threat to human health. One promising alternative for treatment is the application of phage therapy using highly selective bacteriophages. Because of their selectivity, individual screens called phagograms for each patient are required to select phages from a phage library. Phagograms are mostly performed via bacterial cultivation on double layer agar plates and phage addition causing bacterial lysis. However, these assays are work-intensive and have a low ability for parallelization and automation. Hence, highly parallelizable and automatable microbioreactors in the lowest microliter scale could offer an economic solution increasing the throughput of phagograms. This paper demonstrates the applicability of a novel capillary-wave microbioreactor (cwMBR) to perform phagograms. Due to its small volume of only 7 µL and the open-droplet design, it can be easily automated and parallelized in future. Furthermore, the ability of online biomass measurement makes the cwMBR a perfect phagogram platform in the future. Herein, phagograms with <em>E. coli</em> and different concentrations of the phages MM02 and EASG3 were performed as proof of concept for phagograms in the cwMBR. Thereby, the cwMBR was able to measure differences in lysis kinetics of different phages. Furthermore, the phagograms were compared to those in conventional microtiter plate readers revealing the cwMBR as ideal alternative for phagograms as it combines favorable mixing conditions and a phage repellent hydrophilic glass surface with online biomass measurement in an open-droplet design for future parallelization and automation.</p></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1871678424005363/pdfft?md5=4d286a557a4f485ba692e9944dd74ca6&pid=1-s2.0-S1871678424005363-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142056169","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}

{"title":"Exploiting UPO versatility to transform rutin in more soluble and bioactive products","authors":"","doi":"10.1016/j.nbt.2024.08.504","DOIUrl":"10.1016/j.nbt.2024.08.504","url":null,"abstract":"<div><p>The discovery of unspecific peroxygenases (UPOs) completely changed the paradigm of enzyme-based oxyfunctionalization reactions, as these enzymes can transform a wide variety of substrates with a relatively simple reaction mechanism. The fact that UPO can exert both peroxygenative and peroxidative activity in either aromatic or aliphatic carbons, represents a great potential in the production of high value-added products from natural antioxidants. In this work, the flavonoid rutin has been considered as possible substrate for UPO from <em>Agrocybe aegerita</em>, and its peroxygenation or its peroxidation and successive oligomerization have been studied. Different experiments were performed in order to reduce the range of process variables involved and gaining insight on the behavior of this enzyme, leading to a multivariable optimization of UPO-based rutin modification. While trying to preserve enzyme activity this optimization aimed for maximizing the production of more soluble antioxidants. Reusability of the enzyme was evaluated recovering UPO using an enzymatic membrane reactor, revealing challenges in enzyme stability due to inactivation during the filtration stages. The influence of the radical scavenger ascorbic acid on product formation was investigated, revealing its role in directing the reaction towards hydroxylated rutin derivatives, hence indicating a shift towards more soluble and bioactive products.</p></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1871678424005399/pdfft?md5=3e214c5f2d1520a2ab7887b1cb05e856&pid=1-s2.0-S1871678424005399-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142056168","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}

{"title":"Establishment of an RNA-based transient expression system in the green alga Chlamydomonas reinhardtii","authors":"","doi":"10.1016/j.nbt.2024.08.501","DOIUrl":"10.1016/j.nbt.2024.08.501","url":null,"abstract":"<div><p><em>Chlamydomonas reinhardtii</em>, a unicellular green alga, is a prominent model for green biotechnology and for studying organelles’ function and biogenesis, such as chloroplasts and cilia. However, the stable expression of foreign genes from the nuclear genome in <em>C. reinhardtii</em> faces several limitations, including low expression levels and significant differences between clones due to genome position effects, epigenetic silencing, and time-consuming procedures. We developed a robust transient expression system in <em>C. reinhardtii</em> to overcome these limitations. We demonstrated efficient entry of in vitro-transcribed mRNA into wall-less cells and enzymatically dewalled wild-type cells via electroporation. The endogenous or exogenous elements can facilitate efficient transient expression of mRNA in <em>C. reinhardtii</em>, including the 5’ UTR of <em>PsaD</em> and the well-characterized Kozak sequence derived from the <em>Chromochloris zofingiensis</em>. In the optimized system, mRNA expression was detectable in 120 h with a peak around 4 h after transformation. Fluorescently tagged proteins were successfully transiently expressed, enabling organelle labeling and real-time determination of protein sub-cellular localization. Remarkably, transiently expressed IFT46 compensated for the <em>ift46–1</em> mutant phenotype, indicating the correct protein folding and function of IFT46 within the cells. Additionally, we demonstrated the feasibility of our system for studying protein-protein interactions in living cells using bimolecular fluorescence complementation. In summary, the established transient expression system provides a powerful tool for investigating protein localization, function, and interactions in <em>C. reinhardtii</em> within a relatively short timeframe, which will significantly facilitate the study of gene function, genome structure, and green biomanufacturing in <em>C. reinhardtii</em> and potentially in other algae.</p></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1871678424005351/pdfft?md5=e47e31e1c98951d7e7895b1d8654d540&pid=1-s2.0-S1871678424005351-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141996223","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}

{"title":"Enhancing protein productivities in CHO cells through adenosine uptake modulation – Novel insights into cellular growth and productivity regulation","authors":"","doi":"10.1016/j.nbt.2024.08.500","DOIUrl":"10.1016/j.nbt.2024.08.500","url":null,"abstract":"<div><p>Maximizing production potential of recombinant proteins such as monoclonal antibodies (mAbs) in Chinese Hamster Ovary (CHO) cells is a key enabler of reducing cost of goods of biologics. In this study, we explored various strategies to utilize adenosine mediated effects in biologics manufacturing processes. Results show that supplementation of adenosine increases specific productivity by up to two-fold while also arresting cell growth. Introducing adenosine in intensified perfusion processes in a biphasic manner significantly enhanced overall productivity. Interestingly, adenosine effect was observed to be dependent on the cell growth state. Using specific receptor antagonists and inhibitors, we identified that ENTs (primarily Slc29a1) mediate the uptake of adenosine in CHO cell cultures. Transcriptomics data showed an inverse correlation between Slc29a1 expression levels and peak viable cell densities. Data suggests that in fed-batch cultures, adenosine can be produced extracellularly. Blocking Slc29a1 using ENT inhibitors such as DZD and DP alone or in combination with CD73 inhibitor, PSB12379, resulted in a twofold increase in peak viable cell densities as well as productivities in fed batch – a novel strategy that can be applied to biologics manufacturing processes. This is the first study that suggests that adenosine production/accumulation in CHO cell cultures can potentially regulate the transition of CHO cells from exponential to stationary phase. We also demonstrate strategies to leverage this regulatory mechanism to maximize the productivity potential of biologics manufacturing processes.</p></div>","PeriodicalId":19190,"journal":{"name":"New biotechnology","volume":null,"pages":null},"PeriodicalIF":4.5,"publicationDate":"2024-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S187167842400534X/pdfft?md5=e71c2cbd0d460c758df32d65b0e2917b&pid=1-s2.0-S187167842400534X-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141996222","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}