Journal of bioscience and bioengineering最新文献_第6页

Identification of toluene degradation genes in Acinetobacter sp. Tol 5 不动杆菌Tol 5中甲苯降解基因的鉴定。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-11-01 Epub Date: 2025-08-21 DOI: 10.1016/j.jbiosc.2025.07.010

Shogo Yoshimoto , Maiko Hattori , Shori Inoue , Sakura Mori , Yuki Ohara , Katsutoshi Hori

Microbial degradation of aromatic compounds provides sustainable solutions for environmental remediation and bioconversion. Acinetobacter sp. Tol 5 is notable for its strong adhesiveness and potential as a biocatalyst for toluene degradation; however, its toluene metabolic pathway has not been fully elucidated. In this study, genomic analysis identified a cluster of genes in Tol 5 highly similar to the well-known tod operon of Pseudomonas putida, encoding enzymes responsible for toluene metabolism. Phylogenetic analyses indicated that these tod genes, unusual among Acinetobacter species, were likely acquired through horizontal gene transfer. Transcriptomic analyses revealed that todF and todC1 are co-transcribed, while the adjacent fadL2 gene, encoding a putative outer membrane transporter corresponding to P. putida todX, is independently transcribed. Growth experiments using gene-knockout mutants revealed that TodC1, the large subunit of dioxygenase, is essential for growth on toluene, whereas FadL2 is not essential. Growth curves on each carbon source further showed that the todC1 knockout mutant could metabolize benzoate, but not toluene or benzene, confirming that the TOD pathway is the primary route for toluene and benzene degradation in Tol 5. The identification of the functional TOD pathway, which is unique within Acinetobacter, provides genetic and biochemical insights for the development of Tol 5 as an efficient immobilized biocatalyst for the bioremediation and bioconversion of aromatic compounds.

芳香族化合物的微生物降解为环境修复和生物转化提供了可持续的解决方案。不动杆菌sp. Tol 5因其强大的粘附性和作为甲苯降解生物催化剂的潜力而闻名；然而，其甲苯代谢途径尚未完全阐明。在本研究中，基因组分析发现Tol 5中的一组基因与众所周知的恶臭假单胞菌tod操纵子高度相似，编码负责甲苯代谢的酶。系统发育分析表明，这些在不动杆菌物种中不常见的tod基因可能是通过水平基因转移获得的。转录组学分析显示todF和todC1是共转录的，而相邻的fadL2基因是独立转录的，fadL2基因编码一种推定的外膜转运蛋白，对应于恶臭假单胞菌todX。基因敲除突变体的生长实验表明，双加氧酶的大亚基TodC1是在甲苯上生长所必需的，而FadL2不是必需的。各碳源上的生长曲线进一步表明，todC1敲除突变体可以代谢苯甲酸酯，但不能代谢甲苯和苯，证实TOD途径是Tol 5中降解甲苯和苯的主要途径。TOD途径在不动杆菌中具有独特的功能，为Tol 5作为芳香族化合物生物修复和生物转化的高效固定化生物催化剂的开发提供了遗传学和生物化学方面的见解。

{"title":"Identification of toluene degradation genes in Acinetobacter sp. Tol 5","authors":"Shogo Yoshimoto , Maiko Hattori , Shori Inoue , Sakura Mori , Yuki Ohara , Katsutoshi Hori","doi":"10.1016/j.jbiosc.2025.07.010","DOIUrl":"10.1016/j.jbiosc.2025.07.010","url":null,"abstract":"<div><div>Microbial degradation of aromatic compounds provides sustainable solutions for environmental remediation and bioconversion. <em>Acinetobacter</em> sp. Tol 5 is notable for its strong adhesiveness and potential as a biocatalyst for toluene degradation; however, its toluene metabolic pathway has not been fully elucidated. In this study, genomic analysis identified a cluster of genes in Tol 5 highly similar to the well-known <em>tod</em> operon of <em>Pseudomonas putida</em>, encoding enzymes responsible for toluene metabolism. Phylogenetic analyses indicated that these <em>tod</em> genes, unusual among <em>Acinetobacter</em> species, were likely acquired through horizontal gene transfer. Transcriptomic analyses revealed that <em>todF</em> and <em>todC1</em> are co-transcribed, while the adjacent <em>fadL2</em> gene, encoding a putative outer membrane transporter corresponding to <em>P. putida todX</em>, is independently transcribed. Growth experiments using gene-knockout mutants revealed that TodC1, the large subunit of dioxygenase, is essential for growth on toluene, whereas FadL2 is not essential. Growth curves on each carbon source further showed that the <em>todC1</em> knockout mutant could metabolize benzoate, but not toluene or benzene, confirming that the TOD pathway is the primary route for toluene and benzene degradation in Tol 5. The identification of the functional TOD pathway, which is unique within <em>Acinetobacter</em>, provides genetic and biochemical insights for the development of Tol 5 as an efficient immobilized biocatalyst for the bioremediation and bioconversion of aromatic compounds.</div></div>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":"140 5","pages":"Pages 284-289"},"PeriodicalIF":2.9,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144955288","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

Metabolomics-based investigation of the differences between traditional and modern soy sauce 基于代谢组学的传统酱油与现代酱油差异研究。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-11-01 Epub Date: 2025-08-11 DOI: 10.1016/j.jbiosc.2025.07.004

Yoshika Maekawa , Miki Nakamura , Miho Imamura , Eiichiro Fukusaki

Soy sauces can be classified into three categories based on the production method used: honjozo, kongojozo (mixed-brewing, amino acid-decomposed), and kongo (mixed, amino acid-decomposed) soy sauces. Although differences in flavor between traditional (honjozo) and modern (amino acid-decomposed) soy sauces are clear, knowledge of the differences in compound profiles and the relationship between these and the sensory characteristics that affect soy sauce quality is limited. Therefore, this study aimed to investigate the differences between traditional and modern soy sauce compounds using metabolomic analysis, and to investigate the compounds that may be correlated with differences in flavor. Non-targeted gas chromatography/mass spectrometry (GC/MS) metabolomics analysis was performed on nine traditional and six modern soy sauces to annotate 239 soy sauce compounds. Principal component analysis suggested that the production methods used formed clusters that affected the types and amounts of soy sauce compounds, and that the production method had a greater effect on soy sauce composition than did the aging barrels or type of soybean used. Traditional soy sauce was characterized by alcohols and esters, whereas modern soy sauce was characterized by pyrazines. A sensory evaluation revealed that traditional soy sauce was characterized by bitterness and astringency, whereas modern soy sauce was characterized by sweetness and viscosity, suggesting that the method of soy sauce production influences flavor differences. This is the first study to comprehensively characterize the effects of production methods, aging barrels, and soybean types on soy sauce compounds and how these compounds contribute to differences in flavor.

酱油根据制作方法可以分为honjozo、kongojozo（混合酿造、氨基酸分解）和kongo（混合、氨基酸分解）三大类。虽然传统酱油（本酒酱油）和现代酱油（氨基酸分解酱油）在风味上的差异是显而易见的，但对复合结构的差异以及这些差异与影响酱油质量的感官特征之间的关系的了解是有限的。因此，本研究旨在通过代谢组学分析来研究传统酱油和现代酱油化合物之间的差异，并研究可能与风味差异相关的化合物。对9种传统酱油和6种现代酱油进行了非靶向气相色谱/质谱（GC/MS）代谢组学分析，对239种酱油化合物进行了注释。主成分分析表明，不同的生产方式对酱油成分的类型和数量有一定的影响，生产方式对酱油成分的影响大于陈化桶或大豆类型。传统酱油以醇类和酯类为特征，现代酱油以吡嗪类为特征。感官评价结果表明，传统酱油以苦味和涩味为主，而现代酱油以甜味和粘性为主，说明酱油的制作方法影响了风味的差异。这是第一个全面描述生产方法、陈酿桶和大豆类型对酱油化合物的影响以及这些化合物如何导致风味差异的研究。

{"title":"Metabolomics-based investigation of the differences between traditional and modern soy sauce","authors":"Yoshika Maekawa , Miki Nakamura , Miho Imamura , Eiichiro Fukusaki","doi":"10.1016/j.jbiosc.2025.07.004","DOIUrl":"10.1016/j.jbiosc.2025.07.004","url":null,"abstract":"<div><div>Soy sauces can be classified into three categories based on the production method used: <em>honjozo</em>, <em>kongojozo</em> (mixed-brewing, amino acid-decomposed), and <em>kongo</em> (mixed, amino acid-decomposed) soy sauces. Although differences in flavor between traditional (<em>honjozo</em>) and modern (amino acid-decomposed) soy sauces are clear, knowledge of the differences in compound profiles and the relationship between these and the sensory characteristics that affect soy sauce quality is limited. Therefore, this study aimed to investigate the differences between traditional and modern soy sauce compounds using metabolomic analysis, and to investigate the compounds that may be correlated with differences in flavor. Non-targeted gas chromatography/mass spectrometry (GC/MS) metabolomics analysis was performed on nine traditional and six modern soy sauces to annotate 239 soy sauce compounds. Principal component analysis suggested that the production methods used formed clusters that affected the types and amounts of soy sauce compounds, and that the production method had a greater effect on soy sauce composition than did the aging barrels or type of soybean used. Traditional soy sauce was characterized by alcohols and esters, whereas modern soy sauce was characterized by pyrazines. A sensory evaluation revealed that traditional soy sauce was characterized by bitterness and astringency, whereas modern soy sauce was characterized by sweetness and viscosity, suggesting that the method of soy sauce production influences flavor differences. This is the first study to comprehensively characterize the effects of production methods, aging barrels, and soybean types on soy sauce compounds and how these compounds contribute to differences in flavor.</div></div>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":"140 5","pages":"Pages 290-297"},"PeriodicalIF":2.9,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144835200","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

Establishment of genome editing techniques in the marine oleaginous diatom Fistulifera solaris for improved oil accumulation 海洋产油硅藻fisstulifera solaris基因组编辑技术的建立，以改善油脂积累。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-11-01 Epub Date: 2025-08-13 DOI: 10.1016/j.jbiosc.2025.07.008

Satoshi Murata , Natsuno Kushiyama , Yusuke Yabu , Kahori Watanabe , Taiga Fujii , Rein Yasui , Daisuke Nojima , Yoshiaki Maeda , Tomoko Yoshino , Yusuke Matsuda , Tsuyoshi Tanaka

Biofuel production using microalgae has attracted considerable attention owing to high growth rate and lipid accumulation properties. However, further enhancement in lipid productivity is required to render this economically feasible. CRISPR/Cas9, which is one of the powerful genome editing tools, is an essential technique that may solve this problem. The marine diatom Fistulifera solaris JPCC DA0580 is a promising candidate of the biofuel production, since it accumulates significant amount of lipids. However, genome editing techniques have not yet been established for F. solaris, which prevent the construction of valuable strains. In this study, CRISPR/Cas9-mediated specific gene knockout technique was established in F. solaris, through targeting adenine phosphoribosyl transferase gene (apt) and triacylglycerol (TAG) lipase gene (tgl1). Mutations in the target sequence were detected in apt- and tgl1-edited mutants. Moreover, the mutants showed distinct phenotypes, such as suppression of TAG degradation and resistance to 2-fluoroadenine. These results indicate the successful demonstration of CRISPR/Cas9-mediated genome editing in the oleaginous marine diatom F. solaris. Furthermore, oil degradation was successfully suppressed by knocking-out tgl1. The CRISPR/Cas9-mediated genome editing established in this study provides key molecular tools for both the basic biology and the future biotechnological applications of F. solaris, such as biofuel production.

利用微藻生产生物燃料因其高生长速度和油脂积累特性而受到广泛关注。然而，需要进一步提高脂质生产力，使其在经济上可行。CRISPR/Cas9是一种强大的基因组编辑工具，是解决这一问题的关键技术。海洋硅藻Fistulifera solaris JPCC DA0580是一种很有前途的生物燃料生产候选者，因为它积累了大量的脂质。然而，目前还没有针对solaris的基因组编辑技术，这阻碍了有价值菌株的构建。本研究通过靶向腺嘌呤磷酸核糖转移酶基因（apt）和三酰基甘油（TAG）脂肪酶基因（tgl1），建立了CRISPR/ cas9介导的solaris特异性基因敲除技术。在apt-和tgl1编辑的突变体中检测到靶序列的突变。此外，突变体表现出不同的表型，如抑制TAG降解和对2-氟腺嘌呤的抗性。这些结果表明，CRISPR/ cas9介导的基因组编辑在产油的海洋硅藻F. solaris中得到了成功的验证。此外，通过敲除tgl1成功抑制了原油降解。本研究建立的CRISPR/ cas9介导的基因组编辑为solaris的基础生物学和未来生物技术应用（如生物燃料生产）提供了关键的分子工具。

{"title":"Establishment of genome editing techniques in the marine oleaginous diatom Fistulifera solaris for improved oil accumulation","authors":"Satoshi Murata , Natsuno Kushiyama , Yusuke Yabu , Kahori Watanabe , Taiga Fujii , Rein Yasui , Daisuke Nojima , Yoshiaki Maeda , Tomoko Yoshino , Yusuke Matsuda , Tsuyoshi Tanaka","doi":"10.1016/j.jbiosc.2025.07.008","DOIUrl":"10.1016/j.jbiosc.2025.07.008","url":null,"abstract":"<div><div>Biofuel production using microalgae has attracted considerable attention owing to high growth rate and lipid accumulation properties. However, further enhancement in lipid productivity is required to render this economically feasible. CRISPR/Cas9, which is one of the powerful genome editing tools, is an essential technique that may solve this problem. The marine diatom <em>Fistulifera solaris</em> JPCC DA0580 is a promising candidate of the biofuel production, since it accumulates significant amount of lipids. However, genome editing techniques have not yet been established for <em>F. solaris</em>, which prevent the construction of valuable strains. In this study, CRISPR/Cas9-mediated specific gene knockout technique was established in <em>F. solaris</em>, through targeting adenine phosphoribosyl transferase gene (<em>apt</em>) and triacylglycerol (TAG) lipase gene (<em>tgl1</em>). Mutations in the target sequence were detected in <em>apt</em>- and <em>tgl1</em>-edited mutants. Moreover, the mutants showed distinct phenotypes, such as suppression of TAG degradation and resistance to 2-fluoroadenine. These results indicate the successful demonstration of CRISPR/Cas9-mediated genome editing in the oleaginous marine diatom <em>F. solaris</em>. Furthermore, oil degradation was successfully suppressed by knocking-out <em>tgl1</em>. The CRISPR/Cas9-mediated genome editing established in this study provides key molecular tools for both the basic biology and the future biotechnological applications of <em>F. solaris</em>, such as biofuel production.</div></div>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":"140 5","pages":"Pages 271-276"},"PeriodicalIF":2.9,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144846630","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

Enhanced antibody production in Chinese hamster ovary cell cultures supplemented with barley shochu distillation by-product supernatant 添加大麦烧酒蒸馏副产物上清液增强中国仓鼠卵巢细胞抗体产生。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-11-01 Epub Date: 2025-09-01 DOI: 10.1016/j.jbiosc.2025.08.004

Akihiro Nakamura , Masafumi Kadowaki , Kazuya Tsujioka , Takashi Kaieda , Masamichi Kamihira

Antibody production in Chinese hamster ovary (CHO) cell culture was enhanced by supplementing the culture medium with barley shochu distillation by-product supernatant (BX2). To predict antibody production following BX2 addition, fed-batch culture experiments were conducted under varying BX2 conditions using a response surface methodology. BX2 supplementation was predicted to improve antibody production by 138 %, 146 %, 120 %, and 240 % in IgG-producing CHO-MK1, CHO-MK2, CHO-DG44, and Fc-fusion protein-producing CHO-DG44 cells, respectively, compared to controls without BX2. BX2 consisted primarily of 27 % (w/w) bound amino acids, 19 % (w/w) free amino acids, 15 % (w/w) organic acids, 14 % (w/w) glycerol, 14 % (w/w) bound sugars, 6 % (w/w) ash, and 5 % (w/w) free sugars within the soluble solids. BX2 was roughly fractionated into free components (e.g., sugars, amino acids, organic acids, and glycerol fraction) and bound components (e.g., polysaccharides, peptides, peptide-linked sugars, and glycosylated amino acids fraction) using reverse-phase high-performance liquid chromatography. In cultures of IgG-producing CHO-MK2 cells, the addition of BX2 or its bound components enhanced IgG production by 170 % and 117 %, respectively. In contrast, the free components fraction reduced IgG production to 73 % of that observed without BX2. These results suggest that BX2 enhances antibody production through the combined action of multiple components rather than a single component. This study demonstrates that BX2 is an effective additive for enhancing antibody production in CHO cells cultured in high-performance medium typically used in industrial applications.

在培养液中添加大麦烧酒蒸馏副产物上清（BX2），可提高中国仓鼠卵巢（CHO）细胞的抗体产量。为了预测添加BX2后的抗体产量，采用响应面法在不同的BX2条件下进行了补料分批培养实验。与不添加BX2的对照组相比，BX2的补充预计可使产生igg的CHO-MK1、CHO-MK2、CHO-DG44和产生fc融合蛋白的CHO-DG44细胞的抗体产量分别提高138%、146%、120%和240%。BX2主要由27% （w/w）结合氨基酸、19% （w/w）游离氨基酸、15% （w/w）有机酸、14% （w/w）甘油、14% （w/w）结合糖、6% （w/w）灰分和5% （w/w）可溶性固体中的游离糖组成。利用反相高效液相色谱法将BX2大致分为游离组分（如糖、氨基酸、有机酸和甘油部分）和结合组分（如多糖、多肽、肽链糖和糖基化氨基酸部分）。在产生IgG的CHO-MK2细胞的培养中，添加BX2或其结合成分分别使IgG的产生提高了170%和117%。相比之下，游离组分组分将IgG的产量降低到没有BX2时的73%。这些结果表明，BX2通过多种组分的联合作用而不是单一组分来增强抗体的产生。本研究表明，BX2是一种有效的添加剂，可提高CHO细胞在工业应用中常用的高效培养基中产生抗体。

{"title":"Enhanced antibody production in Chinese hamster ovary cell cultures supplemented with barley shochu distillation by-product supernatant","authors":"Akihiro Nakamura , Masafumi Kadowaki , Kazuya Tsujioka , Takashi Kaieda , Masamichi Kamihira","doi":"10.1016/j.jbiosc.2025.08.004","DOIUrl":"10.1016/j.jbiosc.2025.08.004","url":null,"abstract":"<div><div>Antibody production in Chinese hamster ovary (CHO) cell culture was enhanced by supplementing the culture medium with barley shochu distillation by-product supernatant (BX2). To predict antibody production following BX2 addition, fed-batch culture experiments were conducted under varying BX2 conditions using a response surface methodology. BX2 supplementation was predicted to improve antibody production by 138 %, 146 %, 120 %, and 240 % in IgG-producing CHO-MK1, CHO-MK2, CHO-DG44, and Fc-fusion protein-producing CHO-DG44 cells, respectively, compared to controls without BX2. BX2 consisted primarily of 27 % (w/w) bound amino acids, 19 % (w/w) free amino acids, 15 % (w/w) organic acids, 14 % (w/w) glycerol, 14 % (w/w) bound sugars, 6 % (w/w) ash, and 5 % (w/w) free sugars within the soluble solids. BX2 was roughly fractionated into free components (e.g., sugars, amino acids, organic acids, and glycerol fraction) and bound components (e.g., polysaccharides, peptides, peptide-linked sugars, and glycosylated amino acids fraction) using reverse-phase high-performance liquid chromatography. In cultures of IgG-producing CHO-MK2 cells, the addition of BX2 or its bound components enhanced IgG production by 170 % and 117 %, respectively. In contrast, the free components fraction reduced IgG production to 73 % of that observed without BX2. These results suggest that BX2 enhances antibody production through the combined action of multiple components rather than a single component. This study demonstrates that BX2 is an effective additive for enhancing antibody production in CHO cells cultured in high-performance medium typically used in industrial applications.</div></div>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":"140 5","pages":"Pages 341-349"},"PeriodicalIF":2.9,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144955296","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

Locus heterogeneity in the mechanism underlying the superior fermentation performances of phylogenetically distant sake yeasts 系统发育较远的清酒酵母优越发酵性能的基因座异质性机制。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-11-01 Epub Date: 2025-09-02 DOI: 10.1016/j.jbiosc.2025.08.005

Kotaro Mori , Taisuke Seike , Takeshi Akao , Yoshifumi Takao , Tasuku Yamada , Toshinari Takahashi , Fumio Matsuda

Sake yeasts, Saccharomyces cerevisiae strains isolated from Japanese sake fermentation tanks, exhibit superior fermentation performance to that of other yeast strains. Although the exceptional abilities of the modern sake yeast strain K701 have been extensively investigated, those of phylogenetically distant classical sake yeasts remain largely understudied. In this study, we aimed to clarify the mechanism underlying the superior fermentation ability of the classical sake yeast strain Km67 by comparing its genetic and physiological properties with those of the laboratory strains X2180 and K701. Km67 did not exhibit quiescence-specific phenotypes in sake mashes in the same manner as K701. RNA sequencing revealed similar trends between the transcriptomes of Km67 and K701 in sake mash, suggesting that the lack of quiescence entry was related to the higher fermentation ability of Km67. Genetic testing demonstrated that signals upstream of Rim15p were not conveyed downstream, indicating impaired fermentation repression mediated by Rim15p in Km67 cells. Protein phosphatase 2A (PP2A^B55δ) activity declined following CDC55 deletion downstream of Rim15p, consequently reducing the fermentation performance of Km67. However, the extent of fermentation reduction upon CDC55 deletion was not as large as that with X2180 and K701. This suggested the presence of unidentified factors that regulated fermentation independently of PP2A^B55δ or Rim15p in Km67. Thus, our findings demonstrate locus heterogeneity in the mechanism underlying fermentation abilities between the phylogenetically distant sake yeasts Km67 and K701.

从日本清酒发酵罐中分离的酿酒酵母菌，其发酵性能优于其他酵母菌株。尽管现代清酒酵母菌株K701的特殊能力已被广泛研究，但那些在系统发育上遥远的经典清酒酵母菌仍在很大程度上未得到充分研究。本研究旨在通过比较清酒经典酵母菌Km67与实验室菌株X2180和K701的遗传和生理特性，阐明其优越发酵能力的机制。Km67在清酒中没有表现出与K701相同的安静特异性表型。RNA测序结果显示，清酒醪中Km67和K701的转录组具有相似的变化趋势，提示Km67缺乏静息入口与Km67较高的发酵能力有关。基因检测表明，上游的Rim15p信号没有向下游传递，表明在Km67细胞中，Rim15p介导的发酵抑制受损。在Rim15p下游缺失CDC55后，蛋白磷酸酶2A （PP2AB55δ）活性下降，从而降低了Km67的发酵性能。然而，CDC55缺失后发酵还原的程度不如X2180和K701。这表明在Km67中存在不依赖于PP2AB55δ或Rim15p而独立调节发酵的未知因素。因此，我们的研究结果表明，在系统发育上遥远的清酒酵母Km67和K701之间，发酵能力的机制存在基因座异质性。

{"title":"Locus heterogeneity in the mechanism underlying the superior fermentation performances of phylogenetically distant sake yeasts","authors":"Kotaro Mori , Taisuke Seike , Takeshi Akao , Yoshifumi Takao , Tasuku Yamada , Toshinari Takahashi , Fumio Matsuda","doi":"10.1016/j.jbiosc.2025.08.005","DOIUrl":"10.1016/j.jbiosc.2025.08.005","url":null,"abstract":"<div><div>Sake yeasts, <em>Saccharomyces cerevisiae</em> strains isolated from Japanese sake fermentation tanks, exhibit superior fermentation performance to that of other yeast strains. Although the exceptional abilities of the modern sake yeast strain K701 have been extensively investigated, those of phylogenetically distant classical sake yeasts remain largely understudied. In this study, we aimed to clarify the mechanism underlying the superior fermentation ability of the classical sake yeast strain Km67 by comparing its genetic and physiological properties with those of the laboratory strains X2180 and K701. Km67 did not exhibit quiescence-specific phenotypes in sake mashes in the same manner as K701. RNA sequencing revealed similar trends between the transcriptomes of Km67 and K701 in sake mash, suggesting that the lack of quiescence entry was related to the higher fermentation ability of Km67. Genetic testing demonstrated that signals upstream of Rim15p were not conveyed downstream, indicating impaired fermentation repression mediated by Rim15p in Km67 cells. Protein phosphatase 2A (PP2A<sup>B55δ</sup>) activity declined following <em>CDC55</em> deletion downstream of Rim15p, consequently reducing the fermentation performance of Km67. However, the extent of fermentation reduction upon <em>CDC55</em> deletion was not as large as that with X2180 and K701. This suggested the presence of unidentified factors that regulated fermentation independently of PP2A<sup>B55δ</sup> or Rim15p in Km67. Thus, our findings demonstrate locus heterogeneity in the mechanism underlying fermentation abilities between the phylogenetically distant sake yeasts Km67 and K701.</div></div>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":"140 5","pages":"Pages 306-313"},"PeriodicalIF":2.9,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144955388","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

Ribonucleoprotein-based CRISPR/Cas9 genome co-editing in Aspergillus luchuensis mut. kawachii 基于核糖核蛋白的CRISPR/Cas9基因组共编辑在葡曲霉中的应用kawachii。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-11-01 Epub Date: 2025-08-06 DOI: 10.1016/j.jbiosc.2025.07.006

Takefumi Karashima , Ken Oda , Taiki Futagami , Hideki Hokazono , Hideharu Takashita

In this study, we established a ribonucleoprotein-based clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) genome co-editing method for the white koji fungus, Aspergillus luchuensis mut. kawachii. To introduce the single guide RNA-Cas9 ribonucleoprotein complex into protoplast cells of A. luchuensis mut. kawachii, we investigated the conditions for protoplast preparation using Yatalase -Plus-. Subsequently, we employed the ribonucleoprotein-based method to knockout the ATP sulfurylase-encoding sC gene, which imparts selenate resistance in the model strain NBRC 4308 and the industrial strain No. 8046. Furthermore, we explored genome co-editing by simultaneously targeting sC along with either the orotidine 5′-phosphate decarboxylase-encoding pyrG gene or the transcriptional activator of protease genes-encoding prtR gene in NBRC 4308. The transformants were selected in medium containing selenate, resulting in the successful generation of pyrG- and prtR-knockout strains. Similarly, transformants were selected on medium containing selenate, resulting in the successful generation of prtR-knockout strain in No. 8046. These results demonstrate that the ribonucleoprotein-based genome co-editing method is applicable not only to the model strain but also to industrial strains, making it a promising approach for manipulating A. luchuensis mut. kawachii.

在本研究中，我们建立了一种基于核糖核蛋白簇状规则间隔短回文重复序列(CRISPR)/CRISPR相关蛋白9 （Cas9）的白曲真菌Aspergillus luchuensis mut基因组共编辑方法。kawachii。目的：将单导RNA-Cas9核糖核蛋白复合物导入芦杉原生质体细胞。研究了利用Yatalase - plus -制备原生质体的条件。随后，我们采用基于核糖核蛋白的方法敲除了模型菌株NBRC 4308和工业菌株8046中编码ATP硫酰酶的sC基因，该基因赋予了硒酸盐抗性。此外，我们通过同时靶向sC与编码NBRC 4308的orotidine 5'-磷酸脱羧酶pyrG基因或编码prtR基因的蛋白酶基因转录激活因子，探索了基因组共编辑。在含硒酸盐的培养基中选择转化子，成功产生pyg -和prr -敲除菌株。同样，在含有硒酸盐的培养基上选择转化子，成功产生了编号8046的prr敲除菌株。这些结果表明，基于核糖核蛋白的基因组共编辑方法不仅适用于模式菌株，也适用于工业菌株，是一种很有前途的操作方法。kawachii。

{"title":"Ribonucleoprotein-based CRISPR/Cas9 genome co-editing in Aspergillus luchuensis mut. kawachii","authors":"Takefumi Karashima , Ken Oda , Taiki Futagami , Hideki Hokazono , Hideharu Takashita","doi":"10.1016/j.jbiosc.2025.07.006","DOIUrl":"10.1016/j.jbiosc.2025.07.006","url":null,"abstract":"<div><div>In this study, we established a ribonucleoprotein-based clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) genome co-editing method for the white koji fungus, <em>Aspergillus luchuensis</em> mut. <em>kawachii</em>. To introduce the single guide RNA-Cas9 ribonucleoprotein complex into protoplast cells of <em>A. luchuensis</em> mut. <em>kawachii</em>, we investigated the conditions for protoplast preparation using Yatalase -Plus-. Subsequently, we employed the ribonucleoprotein-based method to knockout the ATP sulfurylase-encoding <em>sC</em> gene, which imparts selenate resistance in the model strain NBRC 4308 and the industrial strain No. 8046. Furthermore, we explored genome co-editing by simultaneously targeting <em>sC</em> along with either the orotidine 5′-phosphate decarboxylase-encoding <em>pyrG</em> gene or the transcriptional activator of protease genes-encoding <em>prtR</em> gene in NBRC 4308. The transformants were selected in medium containing selenate, resulting in the successful generation of <em>pyrG</em>- and <em>prtR</em>-knockout strains. Similarly, transformants were selected on medium containing selenate, resulting in the successful generation of <em>prtR</em>-knockout strain in No. 8046. These results demonstrate that the ribonucleoprotein-based genome co-editing method is applicable not only to the model strain but also to industrial strains, making it a promising approach for manipulating <em>A. luchuensis</em> mut. <em>kawachii</em>.</div></div>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":"140 5","pages":"Pages 298-305"},"PeriodicalIF":2.9,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144799199","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

Rapid and practical microbial detection method for brewery hygiene management using the EZ-Fluo system EZ-Fluo系统用于啤酒厂卫生管理的快速实用微生物检测方法。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-11-01 Epub Date: 2025-09-03 DOI: 10.1016/j.jbiosc.2025.08.002

Maika Kitazawa , Nobuchika Takesue , Kazumaru Iijima , Koji Suzuki , Masaki Shimokawa

The early detection of microorganisms in the brewery environment is crucial for taking quick corrective actions and minimizing the risk of microbial contamination. However, hygiene tests using traditional culture methods for visual detection have poor turnaround times. In this study, we optimized the test conditions for the EZ-Fluo Rapid Detection System for brewery hygiene management. In this system, microorganisms are trapped on a membrane filter, cultured on agar, and detected after brief incubation using fluorescence-based technology with a dedicated detector. Among several alternatives, the EZ-Fluo system was chosen for its cost-effectiveness. We determined the appropriate test conditions, including the incubation temperature, fluorescent reagent, and temperature for the fluorescence reaction, using two slow growing environmental indicator species, Acetobacter pasteurianus and Pseudomonas fluorescens. Using the EZ-Fluo system under our optimized detection conditions (hereafter referred to as the EZF-ODC method), nine indicator species common to breweries were detected at a recovery rate of 54.0–117.2 % after only 30 h of incubation. No significant difference was found in colony-forming units among the analysts (p > 0.05). In brewery hygiene tests, the EZF-ODC method (30-h incubation) and the traditional culture method (72-h incubation) yielded comparable detection profiles, with positive or negative results matching in 94.6 % of the 222 samples. The EZF-ODC method can provide results by the following day, representing a significant improvement over the 3 days needed for traditional methods and enabling the rapid implementation of hygiene control measures to maintain a clean brewery environment and possibly expedite production planning.

啤酒厂环境中微生物的早期检测对于采取快速纠正措施和最大限度地减少微生物污染的风险至关重要。然而，使用传统培养方法进行视觉检测的卫生检测周转时间较短。在本研究中，我们优化了用于啤酒厂卫生管理的EZ-Fluo快速检测系统的测试条件。在该系统中，微生物被捕获在膜过滤器上，在琼脂上培养，并在使用专用检测器的荧光技术进行短暂孵育后进行检测。在几个备选方案中，EZ-Fluo系统因其成本效益而被选中。以巴氏醋酸杆菌和荧光假单胞菌两种生长缓慢的环境指示菌为实验对象，确定适宜的实验条件，包括培养温度、荧光试剂和荧光反应温度。在优化的检测条件下（以下简称EZF-ODC法），采用EZ-Fluo系统，仅需30 h，即可检测出9种啤酒常用指示剂，回收率为54.0 ~ 117.2%。分析者之间的菌落形成单位无显著差异（p < 0.05）。在啤酒厂卫生检测中，EZF-ODC法（孵育30小时）和传统培养法（孵育72小时）的检测结果相当，222份样品中有94.6%的阳性或阴性结果匹配。EZF-ODC方法可以在第二天提供结果，这比传统方法需要3天的时间有了显著的改进，并且可以快速实施卫生控制措施，以保持清洁的啤酒厂环境，并可能加快生产计划。

{"title":"Rapid and practical microbial detection method for brewery hygiene management using the EZ-Fluo system","authors":"Maika Kitazawa , Nobuchika Takesue , Kazumaru Iijima , Koji Suzuki , Masaki Shimokawa","doi":"10.1016/j.jbiosc.2025.08.002","DOIUrl":"10.1016/j.jbiosc.2025.08.002","url":null,"abstract":"<div><div>The early detection of microorganisms in the brewery environment is crucial for taking quick corrective actions and minimizing the risk of microbial contamination. However, hygiene tests using traditional culture methods for visual detection have poor turnaround times. In this study, we optimized the test conditions for the EZ-Fluo Rapid Detection System for brewery hygiene management. In this system, microorganisms are trapped on a membrane filter, cultured on agar, and detected after brief incubation using fluorescence-based technology with a dedicated detector. Among several alternatives, the EZ-Fluo system was chosen for its cost-effectiveness. We determined the appropriate test conditions, including the incubation temperature, fluorescent reagent, and temperature for the fluorescence reaction, using two slow growing environmental indicator species, <em>Acetobacter pasteurianus</em> and <em>Pseudomonas fluorescens</em>. Using the EZ-Fluo system under our optimized detection conditions (hereafter referred to as the EZF-ODC method), nine indicator species common to breweries were detected at a recovery rate of 54.0–117.2 % after only 30 h of incubation. No significant difference was found in colony-forming units among the analysts (<em>p</em> > 0.05). In brewery hygiene tests, the EZF-ODC method (30-h incubation) and the traditional culture method (72-h incubation) yielded comparable detection profiles, with positive or negative results matching in 94.6 % of the 222 samples. The EZF-ODC method can provide results by the following day, representing a significant improvement over the 3 days needed for traditional methods and enabling the rapid implementation of hygiene control measures to maintain a clean brewery environment and possibly expedite production planning.</div></div>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":"140 5","pages":"Pages 314-319"},"PeriodicalIF":2.9,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145000633","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

Application of a mitigation method for nitrous oxide emission in a full-scale Carrousel reactor: Carbon footprint assessment 一种减缓氧化亚氮排放方法在全尺寸Carrousel反应堆中的应用：碳足迹评估。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-11-01 Epub Date: 2025-08-21 DOI: 10.1016/j.jbiosc.2025.07.009

Shohei Otomo , Akihiko Terada , Satoru Shibata , Tomoyuki Hori , Eisuke Tamura , Masahiro Ito , Yu-You Li , Fumiaki Takakai , Kunihiro Okano , Naoyuki Miyata , Shuhei Masuda

Nitrous oxide (N₂O) emissions from biological nitrogen removal processes in sewage treatment plants greatly contribute to their overall carbon footprint. The present study aimed to mitigate N₂O emissions from an elliptical Carrousel bioreactor in a full-scale plant. The oxygen supply agitators equipped at the influent point and the opposite side of the reactor was operated alternately. The dissolved N₂O (DN₂O) concentrations were lowered when the agitator at the influent point was suspended while that on the opposite side was running. This scenario was associated with high levels of complementary DNA (RNA) from potential complete denitrifying bacteria, indicating increased N₂O reduction activity utilizing influent organic matter. However, during periods of reduced influent organic load, dissolved oxygen (DO) levels temporarily increased; thereafter, DN₂O increased, accompanied by a decrease in DO. This fluctuation was associated with the accumulation of nitrite and nitrate resulting from ammonia oxidation during the high-DO periods. Based on these findings, an N₂O mitigation strategy was implemented: reducing the oxygen supply and increasing the running time of the opposite-side agitator during the low-organic-loading periods. This approach effectively decreased the DN₂O levels, although a certain degree of instability remained during rainfall events. The median N₂O emission factor decreased from 0.86 % to 0.28 %, reducing the annual carbon footprint of the plant by 14 %. This study provides valuable insights into N₂O mitigation for full-scale plants and demonstrates the great impact of N₂O reduction on their carbon footprint.

污水处理厂生物脱氮过程中产生的氧化亚氮（N2O）排放对其总体碳足迹有很大贡献。本研究旨在减少一氧化二氮排放从一个椭圆形的Carrousel生物反应器在一个完整的工厂。进水点和反应器对面的供氧搅拌器交替运行。当进水点的搅拌器悬停，而另一侧的搅拌器运行时，溶解的N2O （DN2O）浓度降低。这种情况与来自潜在完全反硝化细菌的高水平互补DNA （RNA）有关，表明利用进水有机物的N2O还原活性增加。然而，在降低进水有机负荷期间，溶解氧（DO）水平暂时升高；此后，DN2O升高，DO降低。这种波动与高do期氨氧化引起的亚硝酸盐和硝酸盐积累有关。基于这些发现，实施了一种N2O缓解策略：在低有机负荷期间减少氧气供应并增加对面搅拌器的运行时间。这种方法有效地降低了DN2O水平，尽管在降雨事件期间仍存在一定程度的不稳定性。N2O排放因子中位数从0.86%降至0.28%，使工厂年碳足迹减少14%。本研究为全面植物减少N2O提供了有价值的见解，并证明了N2O减少对其碳足迹的巨大影响。

{"title":"Application of a mitigation method for nitrous oxide emission in a full-scale Carrousel reactor: Carbon footprint assessment","authors":"Shohei Otomo , Akihiko Terada , Satoru Shibata , Tomoyuki Hori , Eisuke Tamura , Masahiro Ito , Yu-You Li , Fumiaki Takakai , Kunihiro Okano , Naoyuki Miyata , Shuhei Masuda","doi":"10.1016/j.jbiosc.2025.07.009","DOIUrl":"10.1016/j.jbiosc.2025.07.009","url":null,"abstract":"<div><div>Nitrous oxide (N<sub>2</sub>O) emissions from biological nitrogen removal processes in sewage treatment plants greatly contribute to their overall carbon footprint. The present study aimed to mitigate N<sub>2</sub>O emissions from an elliptical Carrousel bioreactor in a full-scale plant. The oxygen supply agitators equipped at the influent point and the opposite side of the reactor was operated alternately. The dissolved N<sub>2</sub>O (DN<sub>2</sub>O) concentrations were lowered when the agitator at the influent point was suspended while that on the opposite side was running. This scenario was associated with high levels of complementary DNA (RNA) from potential complete denitrifying bacteria, indicating increased N<sub>2</sub>O reduction activity utilizing influent organic matter. However, during periods of reduced influent organic load, dissolved oxygen (DO) levels temporarily increased; thereafter, DN<sub>2</sub>O increased, accompanied by a decrease in DO. This fluctuation was associated with the accumulation of nitrite and nitrate resulting from ammonia oxidation during the high-DO periods. Based on these findings, an N<sub>2</sub>O mitigation strategy was implemented: reducing the oxygen supply and increasing the running time of the opposite-side agitator during the low-organic-loading periods. This approach effectively decreased the DN<sub>2</sub>O levels, although a certain degree of instability remained during rainfall events. The median N<sub>2</sub>O emission factor decreased from 0.86 % to 0.28 %, reducing the annual carbon footprint of the plant by 14 %. This study provides valuable insights into N<sub>2</sub>O mitigation for full-scale plants and demonstrates the great impact of N<sub>2</sub>O reduction on their carbon footprint.</div></div>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":"140 5","pages":"Pages 332-340"},"PeriodicalIF":2.9,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144955371","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

Enhanced heparosan biosynthesis in Escherichia coli Nissle 1917 through carbon flux redirection 通过碳通量重定向增强大肠杆菌Nissle 1917中肝磷脂的生物合成。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-10-01 Epub Date: 2025-08-05 DOI: 10.1016/j.jbiosc.2025.07.005

Fangqi Shao , Ruiji Wu , Zheng-Jun Li

Heparosan, a critical precursor for heparin production, is naturally biosynthesized as capsular polysaccharides by the probiotic strain Escherichia coli Nissle 1917 (EcN). This study presents a systematic metabolic engineering strategy to enhance heparosan biosynthesis through coordinated pathway engineering and carbon flux redirection. By disrupting glucose catabolism via deletion of zwf and pfkAB, we decoupled cell growth from heparosan synthesis while maintaining precursor availability, elevating titers from 137.68 mg/L to 422.11–486.13 mg/L in mixed carbon source cultures. Subsequent overexpression of UDP-glucose dehydrogenase, a key enzyme in UDP-glucuronic acid biosynthesis, achieved 1.04 g/L heparosan in shake-flask cultivations. Scale-up in a 5-L bioreactor demonstrated industrial scalability, yielding 4.34 g/L heparosan. Our work establishes EcN as a microbial chassis for glycosaminoglycan production and provides a generalizable framework for engineering complex polysaccharide biosynthesis through rational metabolic partitioning.

肝素聚糖是肝素生产的关键前体，是由大肠杆菌Nissle 1917 （EcN）益生菌天然合成的荚膜多糖。本研究提出了一种系统的代谢工程策略，通过协调的途径工程和碳通量重定向来促进肝磷脂的生物合成。通过删除zwf和pfkAB来破坏葡萄糖分解代谢，我们在保持前体可用性的同时，将细胞生长与肝磷脂合成分离，将混合碳源培养的滴度从137.68 mg/L提高到422.11-486.13 mg/L。随后，在摇瓶培养中，udp -葡萄糖醛酸生物合成的关键酶udp -葡萄糖脱氢酶的过表达量达到1.04 g/L。在5升生物反应器中进行放大，证明了工业可扩展性，产率为4.34 g/L。我们的工作建立了EcN作为糖胺聚糖生产的微生物基础，并通过合理的代谢分配为复杂多糖的工程生物合成提供了一个可推广的框架。

引用次数: 0

Solid-state cultivation of Aspergillus oryzae using insoluble plant cell wall polysaccharides and expression analyses of plant polysaccharide degradation-related enzymes 利用不溶性植物细胞壁多糖固态培养米曲霉及植物多糖降解相关酶的表达分析。

IF 2.9 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of bioscience and bioengineering

Pub Date : 2025-10-01 Epub Date: 2025-07-15 DOI: 10.1016/j.jbiosc.2025.06.013

Tomohiko Matsuzawa , Masakazu Ishikawa , Shimma Fujiwa , Naoki Shimada , Hiroshi Kanzaki

Aspergillus oryzae produces multiple plant polysaccharide degradation-related enzymes, which make an important contribution to brewing and biotechnology. In this study, to clarify the mechanisms underlying the enzymatic degradation of plant cell wall polysaccharides by A. oryzae, we prepared insoluble plant cell wall polysaccharides from two plants, mung bean hypocotyls and Chinese chives, and used these as substrates for solid-state cultivation of this fungus. A. oryzae secretes numerous types of carbohydrate-active enzymes that are involved in the degradation of cellulosic and hemicellulosic polysaccharides among insoluble plant cell wall polysaccharides. Transcriptomic analyses revealed that genes encoding both characterized and uncharacterized carbohydrate-active enzymes were highly expressed in solid-state cultures using insoluble plant cell wall polysaccharides, indicating that these enzymes cooperatively degrade plant cell wall polysaccharides.

米曲霉生产多种与植物多糖降解相关的酶，对酿酒和生物技术有重要贡献。本研究为阐明a.m oryzae酶降解植物细胞壁多糖的机制，从绿豆下胚轴和韭菜两种植物中制备了不溶性植物细胞壁多糖，并将其作为固体培养该真菌的底物。在不溶性植物细胞壁多糖中，米曲霉分泌多种类型的碳水化合物活性酶，参与纤维素和半纤维素多糖的降解。转录组学分析显示，在使用不溶性植物细胞壁多糖的固态培养物中，编码特征和非特征糖活性酶的基因都高度表达，表明这些酶协同降解植物细胞壁多糖。

{"title":"Solid-state cultivation of Aspergillus oryzae using insoluble plant cell wall polysaccharides and expression analyses of plant polysaccharide degradation-related enzymes","authors":"Tomohiko Matsuzawa , Masakazu Ishikawa , Shimma Fujiwa , Naoki Shimada , Hiroshi Kanzaki","doi":"10.1016/j.jbiosc.2025.06.013","DOIUrl":"10.1016/j.jbiosc.2025.06.013","url":null,"abstract":"<div><div><em>Aspergillus oryzae</em> produces multiple plant polysaccharide degradation-related enzymes, which make an important contribution to brewing and biotechnology. In this study, to clarify the mechanisms underlying the enzymatic degradation of plant cell wall polysaccharides by <em>A. oryzae</em>, we prepared insoluble plant cell wall polysaccharides from two plants, mung bean hypocotyls and Chinese chives, and used these as substrates for solid-state cultivation of this fungus. <em>A. oryzae</em> secretes numerous types of carbohydrate-active enzymes that are involved in the degradation of cellulosic and hemicellulosic polysaccharides among insoluble plant cell wall polysaccharides. Transcriptomic analyses revealed that genes encoding both characterized and uncharacterized carbohydrate-active enzymes were highly expressed in solid-state cultures using insoluble plant cell wall polysaccharides, indicating that these enzymes cooperatively degrade plant cell wall polysaccharides.</div></div>","PeriodicalId":15199,"journal":{"name":"Journal of bioscience and bioengineering","volume":"140 4","pages":"Pages 200-210"},"PeriodicalIF":2.9,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144649582","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