Journal of bioscience and bioengineering最新文献

Conversion of arachidonic acid into 14,15,19-trihydroxyeicosa-5,8,11-trienoic acid by Torula dematia NBRC 6213. 花生四烯酸转化为14,15,19-三羟基二糖-5,8,11-三烯酸的研究。

IF 2.9 4区生物学

Journal of bioscience and bioengineering Pub Date : 2025-10-07 DOI: 10.1016/j.jbiosc.2025.09.004

Wataru Fujii, Michiki Takeuchi, Si-Bum Park, Kazuki Yagi, Shunta Nakamura, Ei-Tora Yamamura, Jun Ogawa

{"title":"Conversion of arachidonic acid into 14,15,19-trihydroxyeicosa-5,8,11-trienoic acid by Torula dematia NBRC 6213.","authors":"Wataru Fujii, Michiki Takeuchi, Si-Bum Park, Kazuki Yagi, Shunta Nakamura, Ei-Tora Yamamura, Jun Ogawa","doi":"10.1016/j.jbiosc.2025.09.004","DOIUrl":"https://doi.org/10.1016/j.jbiosc.2025.09.004","url":null,"abstract":"Lipids are one of the three major nutrients that play important roles as sources of energy and as components of cell membranes. Fatty acid metabolites exhibit physiological activities. Among fatty acid metabolites, some hydroxy arachidonic acids (ARAs) have bioactive functions. In this study, we found that Torula dematia NBRC 6213 converts ARA into unknown fatty acids. Liquid chromatography-mass spectrometry (LC-MS), gas chromatography-mass spectrometry, and nuclear magnetic resonance (NMR) analyses were conducted to identify this unknown fatty acid. The unknown fatty acid was identified as 14,15,19-trihydroxyeicosa-5,8,11-trienoic acid. Its production was maximized when T. dematia was cultivated in potato dextrose broth (PDB) medium and reacted for 12 h at pH 7.0 and 28 °C. In addition, multiple intermediates were formed during the conversion of ARA to 14,15,19-trihydroxyeicosa-5,8,11-trienoic acid. LC-MS analysis revealed molecular weights of 320, 336, and 338. This suggests that ARA conversion occurs via the hydroxylation, epoxidation, and hydrolysis of the epoxy group. T. dematia also converts other unsaturated fatty acids into similarly oxidized fatty acids.","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2025-10-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145251234","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

Characterization of a molting-related chitinase from a land crab, Chiromantes haematocheir. 一种陆地蟹（Chiromantes haematocheir）几丁质酶的特性。

IF 2.9 4区生物学

Journal of bioscience and bioengineering Pub Date : 2025-10-07 DOI: 10.1016/j.jbiosc.2025.09.006

Yuma Nagakura, Katsuhide Miyake

{"title":"Characterization of a molting-related chitinase from a land crab, Chiromantes haematocheir.","authors":"Yuma Nagakura, Katsuhide Miyake","doi":"10.1016/j.jbiosc.2025.09.006","DOIUrl":"https://doi.org/10.1016/j.jbiosc.2025.09.006","url":null,"abstract":"Chitinases play an important role in many biological processes, including molting, digestion, and immunity in crustaceans. This study represents an attempt to apply chitinases in the field of biotechnology, detecting chitinase mRNAs from a land crab, Chiromantes haematocheir, via transcriptome analysis and analyzing their properties. Seven chitinase genes were detected from the RNA-seq data of midgut glands. Among these genes, TRINITY_DN29294 transcripts accounted for virtually all of total expression of the chitinases. The 29294 cDNA contained a 1467 bp open reading frame, coded for 488 amino acid residues, and was classified into the GH18 chitolectin chitotriosidase and the group 3 crab chitinase. The expression of the 29294-chitinase mRNA was detected in all tissues, with the highest levels expressed in the midgut glands. The transcripts increased significantly in the early post-molted crab compared to the non-molting crab. These results suggest that 29294-chitinase plays important roles in the molting process. While the recombinant 29294-chitinase over-produced in Escherichia coli did not show any activity, the enzyme expressed in Pichia pastoris exhibited sufficient activity. The 29294-chitinase had its optimal pH 3.0. The optimal temperature was relatively high at 45 °C. The enzyme hydrolyzed both soluble and crystalline substrates.","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2025-10-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145251253","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 of bispecific antibody Fc region employing a shark-human chimeric and asymmetric format. 采用鲨鱼-人嵌合和不对称格式设计双特异性抗体Fc区。

IF 2.9 4区生物学

Journal of bioscience and bioengineering Pub Date : 2025-10-01 DOI: 10.1016/j.jbiosc.2025.09.001

Ryota Munetomo, Aiko Inoue, Masayoshi Onitsuka

{"title":"Design of bispecific antibody Fc region employing a shark-human chimeric and asymmetric format.","authors":"Ryota Munetomo, Aiko Inoue, Masayoshi Onitsuka","doi":"10.1016/j.jbiosc.2025.09.001","DOIUrl":"https://doi.org/10.1016/j.jbiosc.2025.09.001","url":null,"abstract":"Bispecific antibodies (BsAbs) can bind to two antigens simultaneously and have undergone rapid advancements in recent years owing to their ability to enable novel mechanisms of action that are unachievable using conventional monoclonal antibodies (mAbs). However, the structural complexity of BsAbs remains a problem during product development. One of these problems is the presence of impurities and by-products. Although BsAbs with the human Fc region must be assembled using heterogeneous polypeptide chains, undesired by-products from unpaired and mispaired chain components can contaminate them. These by-products are difficult to remove in the purification process because their physicochemical properties resemble those of the target BsAb with correct pairing. Here, we designed a novel Fc region for enhanced BsAbs in which the human CH2 domain on one side of the Fc region was replaced with the C2 domain from an immunoglobulin new antigen receptor (IgNAR) shark antibody. The designed BsAbs with chimeric and asymmetric Fcs exhibited separate pH elution profiles against soluble aggregates in protein A affinity chromatography. An overlapping elution profile corresponding to the by-product homogeneous chain observed in human Fc BsAbs was not detected in shark C2-introduced BsAbs. Although another homogeneous by-product was observed in the designed BsAb, introducing N-glycosylation at C2 significantly improved this problem. Additionally, BsAbs with the designed Fc demonstrated higher stability in both the colloidal and structural aspects. This study is the first approach for the chimeric and asymmetric design of Fc using a shark-derived constant domain and offers a novel alternative for BsAb development.","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145212693","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

Development of a method for quantifying metabolites in Escherichia coli colonies using hyperspectral imaging. 利用高光谱成像技术定量大肠杆菌菌落代谢物方法的建立。

IF 2.9 4区生物学

Journal of bioscience and bioengineering Pub Date : 2025-10-01 DOI: 10.1016/j.jbiosc.2025.09.005

Manami Takama, Takatoshi Suematsu, Takayuki Okano, Shumpei Asamizu, Takahiro Bamba, Tomohisa Hasunuma

{"title":"Development of a method for quantifying metabolites in Escherichia coli colonies using hyperspectral imaging.","authors":"Manami Takama, Takatoshi Suematsu, Takayuki Okano, Shumpei Asamizu, Takahiro Bamba, Tomohisa Hasunuma","doi":"10.1016/j.jbiosc.2025.09.005","DOIUrl":"https://doi.org/10.1016/j.jbiosc.2025.09.005","url":null,"abstract":"Fermentation by microorganisms has attracted attention for the synthesis of bulk and fine chemicals with high added value, including pharmaceutical intermediates. To accelerate the development of high-producing microbial strains, a rapid screening method is warranted. This study aimed to develop a novel, nondestructive approach to quantify metabolite production in microbial colonies using hyperspectral imaging (HSI). As a model, we examined the heterologous production of 1,3,5-trihydroxyanthraquinone (AQ256), an anthraquinone with antimicrobial and anticancer activities, using Escherichia coli. Fluorescence spectral data from HSI, along with AQ256 concentrations measured via high-performance liquid chromatography, were used to construct regression models. In addition, red-green-blue (RGB)-based models were developed, as AQ256 exhibits a characteristic reddish-brown color. Four regression models were compared: multiple linear regression, partial least squares regression (PLSR), support vector regression, and random forest regression. Among them, the PLSR model based on HSI data showed the highest prediction accuracy (R2 = 0.75 ± 0.23, root mean square error = 0.08 ± 0.02, mean absolute error = 0.07 ± 0.02). In particular, it outperformed the RGB-based model in extrapolation beyond the training data. These findings demonstrate that the HSI-based method enables accurate, nondestructive quantification of metabolites and has strong potential for high-throughput screening of microbial strains that produce various valuable compounds at elevated yields.","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145212752","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

Importance rapid initial decline in oxidation-reduction potential, followed by an increase in extracellular electron transport activities, for the rapid onset of indigo reduction. 重要的是氧化还原电位的快速初始下降，随后是细胞外电子传递活动的增加，对靛蓝还原的快速开始。

IF 2.9 4区生物学

Journal of bioscience and bioengineering Pub Date : 2025-09-29 DOI: 10.1016/j.jbiosc.2025.08.008

Nowshin Farjana, Hiromitsu Furukawa, Kensuke Igarashi, Souichiro Kato, Isao Yumoto

{"title":"Importance rapid initial decline in oxidation-reduction potential, followed by an increase in extracellular electron transport activities, for the rapid onset of indigo reduction.","authors":"Nowshin Farjana, Hiromitsu Furukawa, Kensuke Igarashi, Souichiro Kato, Isao Yumoto","doi":"10.1016/j.jbiosc.2025.08.008","DOIUrl":"https://doi.org/10.1016/j.jbiosc.2025.08.008","url":null,"abstract":"In most complex microbial systems, the ideal process underlying transitional microbial changes that lead to the formation of functional states is not fully elucidated. To understand the basis for the occurrence of indigo reduction, we analyzed the prerequisites causing transitional shifts in microflora that lead to the indigo-reducing state. To this end, timing of wheat bran (WB) addition, during indigo fermentation process using sukumo (composted leaves of Polygonum tinctorium L.) as the inoculum, substrate, and indigo source, were varied. Early initiation of indigo reduction was achieved through the early proliferation of obligate anaerobic Alkalicella caledoniensis followed by Alkalibacterium spp. or Evansella vedderi. Although it can be predicted that Alkalicella caledoniensis exhibits extracellular electron transport (EET) activity, to promote even effective reduction of indigo, Alkalibacterium spp. or E. vedderi, which have the EET gene sequence series and exert strong metabolic abilities, should emerge using WB. The emergence of Alkalicella caledoniensis was associated with drastic a decrease in bacterial diversity and a concurrent rapid decline in oxidation-reduction potential (ORP). The rate and extent of Alkalicella caledoniensis appearance depended on the rate of ORP reduction. Multivariate analysis (i.e., RDA) revealed that Alkalicella caledoniensis directed the initial drastic changes of microbiota, aligning with the decline in ORP. Prior to these major microbial shifts oxygen consumption by aerobic bacteria utilizing sukumo initiated the ORP decrease. These findings contribute to understanding the approach to steer the initially highly diverse bacterial community during early fermentation toward rapid induction of indigo reduction.","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2025-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145199487","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

Enrichment of soy protein-derived peptides that decrease pancreatic lipase activity using heat-treated porous silica gel and their relationship with bile acid binding activity. 利用热处理多孔硅胶富集降低胰脂肪酶活性的大豆蛋白衍生肽及其与胆汁酸结合活性的关系。

IF 2.9 4区生物学

Journal of bioscience and bioengineering Pub Date : 2025-09-29 DOI: 10.1016/j.jbiosc.2025.09.002

Yusuke Ishii, Yuta Matsunaga, Hirokazu Akiyama, Kazunori Shimizu, Hiroyuki Honda

{"title":"Enrichment of soy protein-derived peptides that decrease pancreatic lipase activity using heat-treated porous silica gel and their relationship with bile acid binding activity.","authors":"Yusuke Ishii, Yuta Matsunaga, Hirokazu Akiyama, Kazunori Shimizu, Hiroyuki Honda","doi":"10.1016/j.jbiosc.2025.09.002","DOIUrl":"https://doi.org/10.1016/j.jbiosc.2025.09.002","url":null,"abstract":"Excessive lipid absorption is a key factor in obesity. Lipids are solubilized in the gut via bile acid (BA) micelles, where pancreatic lipase hydrolyzes them for absorption. This study aimed to enrich pancreatic lipase inhibitory (PLI) peptides from food protein hydrolysates and clarify their inhibition mechanisms. We used heat-treated porous silica gel (HTSG) to selectively enrich basic and hydrophobic peptides through adsorption-desorption. While HTSG has previously enriched PLI peptides, the mechanism remained unclear. Since basic and hydrophobic peptides can bind strongly to BAs like taurocholic acid, we explored their BA-binding and PLI activities. Pepsin hydrolysates from casein, soybean, pea, and rice endosperm were tested with 1 mM sodium taurocholate (TCA). TCA increased lipase activity over 2.5-fold. Soybean pepsin hydrolysate (SPH) showed notable PLI activity, further enhanced approxiamtely 3-fold after HTSG treatment (SPH (after)). LC-MS/MS of SPH (after) identified 1461 peptides. Among 38 high-abundance peptides (Z ≥ 2) chemically synthesized, 9 inhibited pancreatic lipase in the presence of TCA. BA-binding activity was assessed via micelle disruption. Seven of the nine peptides disrupted over 50 % of micelles. Docking simulation was conducted and peptides that exhibited PLI activity even without TCA and showed TCA-binding activity were predicted to bind directly to pancreatic lipase. In summary, we identified 9 PLI peptides from SPH, most of which inhibit pancreatic lipase by binding to BAs. HTSG-based enrichment offers a promising strategy to obtain bioactive peptides that may serve as functional ingredients for obesity prevention.","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2025-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145199543","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

Whole-genome draft assemblies of Paracoccus pantotrophus DSM 11073 and Paracoccus sp. AS002: Phylogenetics entails classification as Paracoccus versutus AS002. 泛养副球菌DSM 11073和副球菌sp. AS002的全基因组草图组装：系统发育需要分类为反副球菌AS002。

IF 2.9 4区生物学

Journal of bioscience and bioengineering Pub Date : 2025-09-27 DOI: 10.1016/j.jbiosc.2025.09.003

Upasana Pal, Denise Bachmann, Linda Fenske, Lars Mathias Blank, Till Tiso

{"title":"Whole-genome draft assemblies of Paracoccus pantotrophus DSM 11073 and Paracoccus sp. AS002: Phylogenetics entails classification as Paracoccus versutus AS002.","authors":"Upasana Pal, Denise Bachmann, Linda Fenske, Lars Mathias Blank, Till Tiso","doi":"10.1016/j.jbiosc.2025.09.003","DOIUrl":"https://doi.org/10.1016/j.jbiosc.2025.09.003","url":null,"abstract":"The Gram-stain-negative bacterium Paracoccus spp., from the class Alphaproteobacteria and family Rhodobacteraceae, exhibits exceptional metabolic flexibility, diverse substrate utilization, and tolerance to abiotic stressors. To broaden the biotechnological applications of the genus, comprehensive sequencing, phylogenetic, and physiological characterization were performed between two strains of the genus, Paracoccus pantotrophus DSM 11073 and Paracoccus sp. AS002. Illumina sequencing yielded a total genome size of 4.2 Mbp for P. pantotrophus DSM 11073 and 4.8 Mbp for Paracoccus sp. AS002. Through phylogenetic analysis using the EDGAR software, Paracoccus sp. AS002 shared with Paracoccus versutus DSM 582 the same clade in the phylogenetic tree and an ANI score of 98.9 % indicating that Paracoccus sp. AS002 could be reclassified as P. versutus AS002. The study was extended to compare various attributes of the sequenced genomes and highlight the metabolic versatility of the genus Paracoccus. The use of a wide substrate panel demonstrated the metabolic versatility of the strains, including the PET monomer ethylene glycol, the C1 carbon source formic acid, and renewable carbon sources such as ethanol. Additionally, the ability of the strains to produce bioplastic was assessed, with P. pantotrophus DSM 11073 producing 36 % and P. versutus AS002 28 % polyhydroxybutyrate (% cell dry weight) on glucose, and 40 % and 16 % on 60 mM ethylene glycol, respectively. This study demonstrates the value of sequencing bacterial strains for biotechnological applications and highlights EDGAR's effectiveness in phylogenetic analysis, paving the way for using Paracoccus as a microbial chassis in sustainable biotechnological processes supporting the circular bioeconomy.","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2025-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145185898","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

Identification and engineering of a cellobiose transporter KmStl1p to enhance cellobiose utilization in Kluyveromyces marxianus and Saccharomyces cerevisiae. 纤维素二糖转运蛋白KmStl1p的鉴定与工程设计以提高马氏克卢维菌和酿酒酵母对纤维素二糖的利用。

IF 2.9 4区生物学

Journal of bioscience and bioengineering Pub Date : 2025-09-20 DOI: 10.1016/j.jbiosc.2025.08.009

Satoshi Ebe, Takuya Abe, Shogo Motozono, Tomoya Kagawa, Riko Kobayashi, Yuki Terauchi, Rinji Akada, Hisashi Hoshida

{"title":"Identification and engineering of a cellobiose transporter KmStl1p to enhance cellobiose utilization in Kluyveromyces marxianus and Saccharomyces cerevisiae.","authors":"Satoshi Ebe, Takuya Abe, Shogo Motozono, Tomoya Kagawa, Riko Kobayashi, Yuki Terauchi, Rinji Akada, Hisashi Hoshida","doi":"10.1016/j.jbiosc.2025.08.009","DOIUrl":"https://doi.org/10.1016/j.jbiosc.2025.08.009","url":null,"abstract":"The yeast Kluyveromyces marxianus, a yeast known for its ability to ferment ethanol at high temperatures, can utilize various sugars including cellobiose, lactose and xylose. This study focused on improving cellobiose utilization by identifying and engineering a cellobiose transporter in K. marxianus. To assess cellobiose utilization capabilities, K. marxianus strains were grown in a cellobiose medium. The strains showed various growth levels, for example, the NCYC2791 strain grew well, while the DMKU3-1042 strain did not. This difference provided a basis for identifying a cellobiose transporter. Thirteen transporter candidate genes from the NCYC2791 genome were expressed in DMKU3-1042. As a result, KmSTL1 overexpression enhanced cell growth in a cellobiose medium. In addition, its disruption in NCYC2791 caused growth defects. To confirm its function, KmSTL1 was co-expressed with a β-glucosidase gene in Saccharomyces cerevisiae EBY.VW1000, which only uptake maltose. This engineered strain grew in cellobiose medium, indicating that KmSTL1 encodes a cellobiose transporter. Expression of GFP-fused KmStl1p in K. marxianus revealed that KmStl1p localized on cell membrane under cellobiose conditions, but was degraded in glucose conditions, suggesting that the transporter is regulated by available sugars. By individually disrupting seven α-arrestin genes in K. marxianus, KmRog3p was identified as a major ubiquitination mediator for KmStl1p degradation. Deletion analysis of KmStl1p revealed that its C-terminus is crucial for recognition by KmRog3p. Furthermore, expressing KmStl1p C-terminus mutants enhanced cellobiose assimilation in both K. marxianus and S. cerevisiae. These findings demonstrate that engineering KmStl1p is an effective strategy to improve cellobiose utilization in yeasts.","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2025-09-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145113215","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

Optimization of heterologous protein production process using genetically engineered Ogataea minuta toward industrial-scale manufacturing. 面向工业规模生产的异源蛋白生产工艺的基因工程优化。

IF 2.9 4区生物学

Journal of bioscience and bioengineering Pub Date : 2025-09-08 DOI: 10.1016/j.jbiosc.2025.08.006

Masashi Tsuda, Yuki Nakatani, Satoshi Baba, Koichi Nonaka, Takehiko Yoko-O, Yasunori Chiba

{"title":"Optimization of heterologous protein production process using genetically engineered Ogataea minuta toward industrial-scale manufacturing.","authors":"Masashi Tsuda, Yuki Nakatani, Satoshi Baba, Koichi Nonaka, Takehiko Yoko-O, Yasunori Chiba","doi":"10.1016/j.jbiosc.2025.08.006","DOIUrl":"https://doi.org/10.1016/j.jbiosc.2025.08.006","url":null,"abstract":"We have developed the methylotrophic yeast Ogataea minuta as a useful host for producing heterologous proteins. In this study, a double mutant that lacks the Prb1 protease and alcohol oxidase was generated and applied for heterologous protein production. Upon our optimization of the fermentation conditions, such as feeding of carbon and nitrogen sources and pH control, this mutant showed increased production of human serum albumin, resulting in a yield of approximately 7.5 g/L at 21 days in the production phase. The established optimal fermentation condition with the double mutant was successfully applied to manufacture a candidate biologic protein: lipocalin derivative which is designed to attach to the calcitonin gene-related peptide, on an industrial scale. The candidate biologic protein was successfully manufactured at an industrial scale in a 4500 L bioreactor under well-controlled conditions. This first successful case highlights the potential of our O. minuta-based production system, and this study is helpful for a large-scale cultivation of a methylotrophic yeast for protein production.","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2025-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145029943","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

Sterilizable autoantigen immobilized column platform for broad-spectrum removal of pathogenic autoantibodies in autoimmune diseases. 可灭菌的自身抗原固定化柱平台，用于自身免疫性疾病中病原性自身抗体的广谱去除。

IF 2.9 4区生物学

Journal of bioscience and bioengineering Pub Date : 2025-09-04 DOI: 10.1016/j.jbiosc.2025.08.007

Midori Futami, Eri Kurozumi, Masaya Kamo, Soudai Taguchi, Tomoaki Nakai, Junichiro Futami

{"title":"Sterilizable autoantigen immobilized column platform for broad-spectrum removal of pathogenic autoantibodies in autoimmune diseases.","authors":"Midori Futami, Eri Kurozumi, Masaya Kamo, Soudai Taguchi, Tomoaki Nakai, Junichiro Futami","doi":"10.1016/j.jbiosc.2025.08.007","DOIUrl":"https://doi.org/10.1016/j.jbiosc.2025.08.007","url":null,"abstract":"Blood purification using immunoadsorbent columns is a therapeutic strategy for removing pathogenic autoantibodies in autoimmune diseases. Currently available columns have limitations: Trp/Phe columns offer cost-effectiveness and sterilizability, but lack antigen specificity and have limited capacity to remove diverse pathogenic autoantibodies; whereas Protein A/peptide/anti-human IgG columns target all antibodies, regardless of pathogenicity, limiting specificity, and often require sterile production due to low stability under sterilization conditions, except for peptide ligands. Full-length autoantigen-immobilized immunoadsorbent columns have great potential to specifically adsorb targeted autoantibodies, because autoantibodies recognize diverse epitopes that vary among individuals. However, it is challenging to prepare biologically active autoantigens on a large scale and maintain the quality of antigen-immobilized columns after sterilization. This study introduced a novel approach for preparing sterilizable antigen-immobilized columns that target autoantibodies, excluding those with conformational epitope specificity. Two type I transmembrane protein-coding extracellular domains associated with autoimmunity and their rabbit antisera were used as models. Recombinant human contactin-associated protein-like 2 (Caspr2) and muscle-specific tyrosine-protein kinase receptor (MuSK) were expressed as bacterial inclusion bodies. These compounds were solubilized and purified using Cys-specific chemical cationization. Columns immobilized with water-soluble S-cationized Caspr2 or MuSK effectively captured specific antibodies from rabbit antisera against each antigen, retaining their capacity after standard sterilization. This approach offers a promising solution for developing immunoadsorbent columns with enhanced specificity and sterilizability and is applicable to various autoantibody-related disorders.","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2025-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145006171","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