Journal of bioscience and bioengineering最新文献_第8页

Enhancing extracellular membrane vesicle productivity of Shewanella vesiculosa HM13, a prospective host for vesiculation-mediated protein secretion, by weakening outer membrane-peptidoglycan linkage 通过削弱外膜-肽聚糖连接，提高囊泡介导蛋白质分泌的潜在宿主 Shewanella vesiculosa HM13 的细胞外膜囊泡生产力。

IF 2.3 4区生物学

Journal of bioscience and bioengineering Pub Date : 2024-05-25 DOI: 10.1016/j.jbiosc.2024.05.005

{"title":"Enhancing extracellular membrane vesicle productivity of Shewanella vesiculosa HM13, a prospective host for vesiculation-mediated protein secretion, by weakening outer membrane-peptidoglycan linkage","authors":"","doi":"10.1016/j.jbiosc.2024.05.005","DOIUrl":"10.1016/j.jbiosc.2024.05.005","url":null,"abstract":"<div>Shewanella vesiculosa HM13, a psychrotrophic gram-negative bacterium isolated from the intestinal contents of horse mackerel, produces abundant extracellular membrane vesicles (EMVs) by budding the outer membrane. The EMVs of this bacterium carry a single major cargo protein, P49, of unknown function, which may be useful as a carrier for the secretory production of heterologous proteins as cargoes of EMVs. In this study, to increase the utility of S. vesiculosa HM13 as a host for EMV-mediated protein production, we improved its EMV productivity by weakening the linkage between the outer membrane and underlying peptidoglycan layer. In gram-negative bacteria, the outer membrane is connected to peptidoglycans predominantly through Braun's lipoprotein (Lpp), and the formation of this linkage is catalyzed by an l,d-transpeptidase (Ldt). We constructed gene-disrupted mutants of Lpp and Ldt and assessed their EMV productivity. The EMVs of the lpp- and ldt-disrupted mutants grown at 18 °C were evaluated using nanoparticle tracking analysis, and their morphologies were observed using transmission electron microscopy. As a result, an approximately 2.5-fold increase in EMV production was achieved, whereas the morphology of the EMVs of these mutants remained almost identical to that of the parent strain. In accordance with the increase in EMV production, the mutants secreted approximately 2-fold higher amounts of P49 than the parent strain into the culture broth as the EMV cargo. These findings will contribute to the development of an EMV-based secretory production system for heterologous proteins using S. vesiculosa HM13 as a host.</div>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":"138 2","pages":"Pages 137-143"},"PeriodicalIF":2.3,"publicationDate":"2024-05-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141155137","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Production of single cell oil by Lipomyces starkeyi from waste plant oil generated by the palm oil mill industry 星形脂霉菌利用棕榈油厂产生的废植物油生产单细胞油。

IF 2.3 4区生物学

Journal of bioscience and bioengineering Pub Date : 2024-05-22 DOI: 10.1016/j.jbiosc.2024.04.005

引用次数: 0

Optimizing in vitro expression balance of central dogma-related genes using parallel reaction monitoring 利用平行反应监测优化中枢教条相关基因的体外表达平衡。

IF 2.3 4区生物学

Journal of bioscience and bioengineering Pub Date : 2024-05-18 DOI: 10.1016/j.jbiosc.2024.04.006

{"title":"Optimizing in vitro expression balance of central dogma-related genes using parallel reaction monitoring","authors":"","doi":"10.1016/j.jbiosc.2024.04.006","DOIUrl":"10.1016/j.jbiosc.2024.04.006","url":null,"abstract":"<div>The creation of a self-replicating synthetic cell is an essential to understand life self-replication. One method to create self-replicating artificial cells is to reconstitute the self-replication system of living organisms in vitro. In a living cell, self-replication is achieved via a system called the autonomous central dogma, a system in which central dogma-related factors are autonomously synthesized and genome replication, transcription, and translation are driven by nascent factors. Various studies to reconstitute some processes of the autonomous central dogma in vitro have been conducted. However, in vitro reconstitution of the entire autonomous central dogma system is difficult as it requires balanced expression of several related genes. Therefore, we developed a method to simultaneously quantify and optimize the in vitro expression balance of multiple genes. First, we developed a quantitative mass spectrometry method targeting genome replication-related proteins as a model of central dogma-related factors and acquired in vitro expression profiles of these genes. Additionally, we demonstrated that the in vitro expression balance of these genes can be easily optimized by adjusting the input gene ratio based on the data obtained by the developed method. This study facilitated the easy optimization of the in vitro expression balance of multiple genes. Therefore, extending the scope of this method to other central dogma-related factors will accelerate attempts of self-replicating synthetic cells creation.</div>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":"138 2","pages":"Pages 97-104"},"PeriodicalIF":2.3,"publicationDate":"2024-05-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140957620","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Achieving unprecedented stability in lyophilized recombinase polymerase amplification with thermostable pyruvate kinase from Thermotoga maritima 利用海洋嗜热菌（Thermotoga maritima）的恒温丙酮酸激酶实现冻干重组酶聚合酶扩增的空前稳定性。

IF 2.8 4区生物学

Journal of bioscience and bioengineering Pub Date : 2024-05-07 DOI: 10.1016/j.jbiosc.2024.04.003

Kevin Maafu Juma , Yuto Murakami , Kenta Morimoto , Teisuke Takita , Kenji Kojima , Koichiro Suzuki , Itaru Yanagihara , Soichiro Ikuta , Shinsuke Fujiwara , Kiyoshi Yasukawa

{"title":"Achieving unprecedented stability in lyophilized recombinase polymerase amplification with thermostable pyruvate kinase from Thermotoga maritima","authors":"Kevin Maafu Juma , Yuto Murakami , Kenta Morimoto , Teisuke Takita , Kenji Kojima , Koichiro Suzuki , Itaru Yanagihara , Soichiro Ikuta , Shinsuke Fujiwara , Kiyoshi Yasukawa","doi":"10.1016/j.jbiosc.2024.04.003","DOIUrl":"10.1016/j.jbiosc.2024.04.003","url":null,"abstract":"<div>Recombinase polymerase amplification (RPA) is an isothermal DNA amplification reaction at around 41 °C using recombinase (Rec), single-stranded DNA-binding protein (SSB), strand-displacing DNA polymerase (Pol), and an ATP-regenerating enzyme. Considering the onsite use of RPA reagents, lyophilized RPA reagents with long storage stability are highly desired. In this study, as one of the approaches to solve this problem, we attempted to use a thermostable pyruvate kinase (PK). PK gene was isolated from a thermophilic bacterium Thermotoga maritima (Tma-PK). Tma-PK was expressed in Escherichia coli and purified from the cells. Tma-PK exhibited higher thermostability than human PK. The purified Tma-PK preparation was applied to RPA as an ATP-regenerating enzyme. Liquid RPA reagent with Tma-PK exhibited the same performance as that with human PK. Lyophilized RPA reagent with Tma-PK exhibited higher performance than that with human PK. Combined with our previous results of RPA reagents of thermostable Pol from a thermophilic bacterium, Aeribacillus pallidus, the results in this study suggest that thermostable enzymes are preferable to mesophilic ones as a component in lyophilized RPA reagents.</div>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":"138 1","pages":"Pages 29-35"},"PeriodicalIF":2.8,"publicationDate":"2024-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140891630","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Double knockout of two target genes via genome co-editing using a nitrate transporter gene nrtA and a putative thiamine transporter gene thiI as selectable markers in Aspergillus oryzae 使用硝酸盐转运体基因 nrtA 和推测的硫胺素转运体基因 thiI 作为 Aspergillusoryzae 中的可选择标记，通过基因组联合编辑实现两个目标基因的双重敲除。

IF 2.8 4区生物学

Journal of bioscience and bioengineering Pub Date : 2024-04-23 DOI: 10.1016/j.jbiosc.2024.03.007

Koichi Tamano , Haruka Takayama

{"title":"Double knockout of two target genes via genome co-editing using a nitrate transporter gene nrtA and a putative thiamine transporter gene thiI as selectable markers in Aspergillus oryzae","authors":"Koichi Tamano , Haruka Takayama","doi":"10.1016/j.jbiosc.2024.03.007","DOIUrl":"10.1016/j.jbiosc.2024.03.007","url":null,"abstract":"<div>Genome co-editing technology is effective in breeding filamentous fungi for applications in the fermentation industry, achieving site-directed mutagenesis, the status of non-genetically modified organisms (non-GMOs), and wild-type-like growth phenotype. Prior to this study, thiI gene was found as a selectable marker for such genome co-editing in the filamentous fungus Aspergillus oryzae, while it cannot be reused via marker recycling. Therefore, we aimed to identify another marker gene to knock out another target gene via genome co-editing in A. oryzae. In this study, we focused on the membrane transporter gene nrtA (AO090012000623), which promotes uptake of nitrate (NO3-). It is known that, in nrtA knockout strain, chlorate (ClO3-), an analog of nitrate with antifungal activity, cannot be imported into the cytosol, which enables the mutant to grow in the presence of chlorate. Based on this information, knockout of the target gene wA was attempted using both nrtA- and wA-specific single-guide RNAs via genome co-editing with KClO3 supplementation in A. oryzae laboratory strain RIB40 and industrial strain KBN616. Resultantly, wA knockout mutant was generated, and nrtA was identified as a selectable marker. Moreover, this genome co-editing system using nrtA was compatible with that using thiI, and thus, a double knockout mutant of two target genes wA and yA was constructed in RIB40 while maintaining non-GMO status and wild-type-like growth. As nrtA homologs have been found in several industrial Aspergillus species, genome co-editing using homolog genes as selectable markers is plausible, which would contribute to the widespread breeding of industrial strains of Aspergilli.</div>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":"138 1","pages":"Pages 36-43"},"PeriodicalIF":2.8,"publicationDate":"2024-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140769868","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

New insights into the characteristic flavor components of traditional sour beers such as Lambic and Flanders Red Ale beers 对传统酸味啤酒（如兰比克啤酒和佛兰德斯红啤酒）的特色风味成分有了新的认识。

IF 2.8 4区生物学

Journal of bioscience and bioengineering Pub Date : 2024-04-22 DOI: 10.1016/j.jbiosc.2024.04.002

Kyoya Onishi , Masahiro Furuno , Asuka Mori , Eiichiro Fukusaki

{"title":"New insights into the characteristic flavor components of traditional sour beers such as Lambic and Flanders Red Ale beers","authors":"Kyoya Onishi , Masahiro Furuno , Asuka Mori , Eiichiro Fukusaki","doi":"10.1016/j.jbiosc.2024.04.002","DOIUrl":"10.1016/j.jbiosc.2024.04.002","url":null,"abstract":"<div>In recent years, the demand for beers with a variety of flavors has increased considerably owing to the diversification of consumer preferences. Sour beer is characterized by a sour taste unlike normal beer flavor, and previous studies on sour beer have been primarily focused on addressing issues, such as inconsistent product quality and long production time, and on the associated microorganisms. Scientific knowledge regarding the characteristic flavor of sour beer and flavor components is limited. Therefore, in this study, we aimed to clarify the characteristic sensory attributes of sour beer and the component profiles that explain these attributes. Component analysis was performed on 10 traditional sour beers (eight Flanders Red Ales and two Lambics), using untargeted gas chromatography-mass spectrometry with liquid–liquid extraction, liquid chromatography-mass spectrometry targeting amines and anionic compounds. Further, sensory evaluation was conducted by well-trained panelists via quantitative descriptive analysis. Orthogonal partial least squares regression analysis was also conducted to investigate candidate flavor components. Thus, 261 components were identified and our methods could explain the flavor attributes of the examined samples. Comprehensive component profiling data also showed that differences in fermentation method, barrel aging duration, and blending ratio affected beer flavor. Further, Lambics were found to be characterized by citrus and phenolic aroma, while Flanders Red Ales were characterized by solvent-like aroma, sourness complexity, full bodied, graininess, astringency, and bitterness. These findings may serve as a basis for addressing issues related to sour beer production and may facilitate process design for obtaining targeted sour beer flavors.</div>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":"138 1","pages":"Pages 54-62"},"PeriodicalIF":2.8,"publicationDate":"2024-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140768223","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Design and validation of functionalized redox-responsive hydrogel beads for high-throughput screening of antibody-secreting mammalian cells 设计和验证用于高通量筛选分泌抗体的哺乳动物细胞的功能化氧化还原反应水凝胶珠。

IF 2.8 4区生物学

Journal of bioscience and bioengineering Pub Date : 2024-04-21 DOI: 10.1016/j.jbiosc.2024.04.001

Diah Anggraini Wulandari , Kyosuke Tsuru , Kosuke Minamihata , Rie Wakabayashi , Go Egami , Yoshinori Kawabe , Masamichi Kamihira , Masahiro Goto , Noriho Kamiya

{"title":"Design and validation of functionalized redox-responsive hydrogel beads for high-throughput screening of antibody-secreting mammalian cells","authors":"Diah Anggraini Wulandari , Kyosuke Tsuru , Kosuke Minamihata , Rie Wakabayashi , Go Egami , Yoshinori Kawabe , Masamichi Kamihira , Masahiro Goto , Noriho Kamiya","doi":"10.1016/j.jbiosc.2024.04.001","DOIUrl":"10.1016/j.jbiosc.2024.04.001","url":null,"abstract":"<div>Antibody drugs play a vital role in diagnostics and therapy. However, producing antibodies from mammalian cells is challenging owing to cellular heterogeneity, which can be addressed by applying droplet-based microfluidic platforms for high-throughput screening (HTS). Here, we designed an integrated system based on disulfide-bonded redox-responsive hydrogel beads (redox-HBs), which were prepared through enzymatic hydrogelation, to compartmentalize, screen, select, retrieve, and recover selected Chinese hamster ovary (CHO) cells secreting high levels of antibodies. Moreover, redox-HBs were functionalized with protein G as an antibody-binding module to capture antibodies secreted from encapsulated cells. As proof-of-concept, cells co-producing immunoglobulin G (IgG) as the antibody and green fluorescent protein (GFP) as the reporter molecule, denoted as CHO(IgG/GFP), were encapsulated into functionalized redox-HBs. Additionally, antibody-secreting cells were labeled with protein L-conjugated horseradish peroxidase using a tyramide amplification system, enabling fluorescence staining of the antibody captured inside the beads. Redox-HBs were then applied to fluorescence-activated droplet sorting, and selected redox-HBs were degraded by reducing the disulfide bonds to recover the target cells. The results indicated the potential of the developed HTS platform for selecting a single cell viable for biopharmaceutical production.</div>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":"138 1","pages":"Pages 89-95"},"PeriodicalIF":2.8,"publicationDate":"2024-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140757189","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Expression of an endo-rhamnogalacturonase from Aspergillus aculeatus enhances release of Arabidopsis transparent mucilage 表达曲霉的内鼠李糖半乳糖醛酸酶可促进拟南芥透明粘液的释放。

IF 2.8 4区生物学

Journal of bioscience and bioengineering Pub Date : 2024-04-20 DOI: 10.1016/j.jbiosc.2024.03.006

Takao Ohashi , Yurika Mabira , Yutaro Mitsuyoshi , Hiroyuki Kajiura , Ryo Misaki , Takeshi Ishimizu , Kazuhito Fujiyama

{"title":"Expression of an endo-rhamnogalacturonase from Aspergillus aculeatus enhances release of Arabidopsis transparent mucilage","authors":"Takao Ohashi , Yurika Mabira , Yutaro Mitsuyoshi , Hiroyuki Kajiura , Ryo Misaki , Takeshi Ishimizu , Kazuhito Fujiyama","doi":"10.1016/j.jbiosc.2024.03.006","DOIUrl":"10.1016/j.jbiosc.2024.03.006","url":null,"abstract":"<div>Mucilage is a gelatinous and sticky hydrophilic polysaccharide released from epidermal cells of seed coat after the hydration of mature seeds and is composed primarily of unbranched rhamnogalacturonan I (RG-I). In this study, we produced a recombinant endo-RG-I hydrolase from Aspergillus aculeatus (AaRhgA) in the fission yeast Schizosaccharomyces pombe and examined its substrate preference for pyridylaminated (PA) RG-I with the various degrees of polymerization (DP). Recombinant AaRhgA requires PA-RG-I with a DP of 10 or higher for its hydrolase activity. We heterologously expressed the AarhgA gene under the strong constitutive promoter, cauliflower mosaic virus 35S promoter, in Arabidopsis thaliana. In a series of biochemical analyses of each mucilage fraction released from the water-imbibed seeds of the transgenic plants, we found the enhanced deposition of the transparent mucilage layer that existed in the peripheral regions of the adherent mucilage and was not stained with ruthenium red. This study demonstrated the feasibility of manipulating the mucilage organization by heterologous expression of the endo-RG-I hydrolase.</div>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":"138 1","pages":"Pages 73-82"},"PeriodicalIF":2.8,"publicationDate":"2024-04-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140775987","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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

Fabrication of a cell culture scaffold that mimics the composition and structure of bone marrow extracellular matrix 模拟骨髓细胞外基质组成和结构的细胞培养支架的制作。

IF 2.8 4区生物学

Journal of bioscience and bioengineering Pub Date : 2024-04-20 DOI: 10.1016/j.jbiosc.2024.03.008

Ayana Yamaguchi , Yoshihide Hashimoto , Jun Negishi

引用次数: 0

Purification, characterization and application of collagenolytic protease from Bacillus subtilis strain MPK 枯草芽孢杆菌 MPK 菌株胶原溶解蛋白酶的纯化、表征和应用。

IF 2.8 4区生物学

Journal of bioscience and bioengineering Pub Date : 2024-04-17 DOI: 10.1016/j.jbiosc.2024.03.003

Madhuri Vijay Bhuimbar , Chidambar Balbhim Jalkute , Prashant Kishor Bhagwat , Padma Babulal Dandge

{"title":"Purification, characterization and application of collagenolytic protease from Bacillus subtilis strain MPK","authors":"Madhuri Vijay Bhuimbar , Chidambar Balbhim Jalkute , Prashant Kishor Bhagwat , Padma Babulal Dandge","doi":"10.1016/j.jbiosc.2024.03.003","DOIUrl":"10.1016/j.jbiosc.2024.03.003","url":null,"abstract":"<div>A new extracellular protease from Bacillus subtilis strain MPK with collagenolytic activity was isolated and purified. Fish skin which otherwise would be treated as waste is used as substrate for the production of protease. Using various techniques such as ammonium sulphate precipitation and ion exchange chromatography, protease was purified and characterized subsequently. Protease of approximately 61 kDa molecular weight was purified by 135.7-fold with 18.42% enzyme recovery. The protease showed effective properties like pH and temperature stability over a broad range with optimum pH 7.5 and temperature 60 °C. Km and Vmax were found to be 1.92 mg ml−1 and 1.02 × 10−4 mol L−1 min−1, respectively. The protease exhibited stability in various ions, surfactants, inhibitors and organic solvents. Subsequently, the protease was successfully utilized for collagen hydrolysis to generate collagen peptides; thus, the produced protease would be a potential candidate for multifaceted applications in food and pharmaceutical industries due to its significant characteristics and collagenolytic properties.</div>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":"138 1","pages":"Pages 21-28"},"PeriodicalIF":2.8,"publicationDate":"2024-04-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140780655","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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