Journal of General and Applied Microbiology最新文献

Diverse fungal communities in commercial karebushi produced with or without a starter culture. 在有或没有发酵剂的情况下，商业鹿茸中不同的真菌群落。

IF 0.8 4区生物学

Journal of General and Applied Microbiology Pub Date : 2025-08-26 DOI: 10.2323/jgam.2025.08.001

Kazuna Yanagi, Kentaro Hiramatsu, Chihiro Kadooka, Atsushi Nishitani, Kayu Okutsu, Yumiko Yoshizaki, Kazunori Takamine, Hisanori Tamaki, Taiki Futagami

引用次数: 0

Characterization of the Azotobacter vinelandii nitrogenase complex expressed in Escherichia coli toward further activity improvement. 在大肠杆菌中表达的氮化固氮菌氮酶复合体的鉴定，以期进一步提高其活性。

IF 0.8 4区生物学

Journal of General and Applied Microbiology Pub Date : 2025-07-15 Epub Date: 2025-01-10 DOI: 10.2323/jgam.2024.12.001

Yusuke Ito, Daisuke Yoshidome, Makoto Hidaka, Yasuko Araki, Kotaro Ito, Saori Kosono, Makoto Nishiyama

引用次数: 0

CRISPR-Cas9 genome editing of miso and soy source yeast Zygosaccharomyces sp. miso和soy来源酵母Zygosaccharomyces sp.的CRISPR-Cas9基因组编辑

IF 0.8 4区生物学

Journal of General and Applied Microbiology Pub Date : 2025-07-15 Epub Date: 2025-04-30 DOI: 10.2323/jgam.2025.04.002

Tomoo Ogata, Kotori Koide, Shiori Kudou, Miu Suto, Kotaro Uehara, Teruya Kaneko

{"title":"CRISPR-Cas9 genome editing of miso and soy source yeast Zygosaccharomyces sp.","authors":"Tomoo Ogata, Kotori Koide, Shiori Kudou, Miu Suto, Kotaro Uehara, Teruya Kaneko","doi":"10.2323/jgam.2025.04.002","DOIUrl":"10.2323/jgam.2025.04.002","url":null,"abstract":"Genome modification would be useful for developing breeding techniques for haploid Zygosaccharomyces rouxii and natural hybrid allodiploid Zygosaccharomyces sp. yeast strains used in miso and soy sauce production. In this study, genome editing using CRISPR-Cas9 was attempted in Zygosaccharomyces sp. strains. Based on techniques in Saccharomyces cerevisiae, the Cas9 gene and guide RNA (gRNA) were expressed from the same plasmid. Targeting of the ZygoLEU2 gene of haploid Z. rouxii strain DA2 led to of a single-nucleotide insertion in the ORF, resulting in termination of translation at 10 amino acids. This single-base insertion was 3-bp upstream of the protospacer-associated motif (PAM) sequence, suggesting that it occurred during the repair process following the Cas9-induced double-strand break. The transformant was auxotrophic for leucine, verifying that genome editing using CRISPR-Cas9 had occurred. Application of the CRISPR-Cas9 system to allodiploid Zygosaccharomyces sp. strains, which have T- and P-subgenomes, resulted in transformants with base insertions or deletions upstream of the PAM sequence, or insertions of different subgenome sequences. Leucine-auxotrophic transformants were obtained in which the ORF of the ZygoLEU2 gene in both subgenomes were mutated. In some genome-edited strains, a significant region of one subgenome chromosome was missing. Lastly, we applied CRISPR-Cas9 to the gene encoding Hog1, a protein kinase involved in adaptation to high-salt and high-osmolarity conditions. Mutation of the HOG1 genes of both the T- and P-subgenomes by CRISPR-Cas9 significantly reduced growth in high salt and high osmolarity conditions.","PeriodicalId":15842,"journal":{"name":"Journal of General and Applied Microbiology","volume":" ","pages":""},"PeriodicalIF":0.8,"publicationDate":"2025-07-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144063999","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 characterization of a methyl-accepting chemotaxis protein in Ralstonia pseudosolanacearum using chemically undefined materials. 利用化学性质不明确的材料鉴定和表征假龙葵中一个甲基接受趋化蛋白。

IF 0.8 4区生物学

Journal of General and Applied Microbiology Pub Date : 2025-07-15 Epub Date: 2025-03-17 DOI: 10.2323/jgam.2025.03.001

Asmaa Ali Ahmed, Akiko Hida, Takahisa Tajima, Junichi Kato

{"title":"Identification and characterization of a methyl-accepting chemotaxis protein in Ralstonia pseudosolanacearum using chemically undefined materials.","authors":"Asmaa Ali Ahmed, Akiko Hida, Takahisa Tajima, Junichi Kato","doi":"10.2323/jgam.2025.03.001","DOIUrl":"10.2323/jgam.2025.03.001","url":null,"abstract":"Ralstonia pseudosolanacearum is a plant-pathogenic bacterium that causes bacterial wilt in economically important crops. Chemotaxis is required for full virulence in R. pseudosolanacearum. R. pseudosolanacearum Ps29 possesses 20 methyl-accepting chemotaxis proteins (MCPs) and 2 MCP-like chemoreceptors. To understand the role of chemotaxis in plant infection, we are characterizing the functions of these 20 MCPs. Out of 20 MCPs, 8 MCPs have been characterized. To characterize the remaining MCPs, we deleted the 8 genes encoding characterized MCPs in R. pseudosolanacearum Ps29 to construct R. pseudosolanacearum D8. R. pseudosolanacearum D8 was examined for chemotactic responses to several chemically undefined materials including vegetable juices and tryptic soy broth (TSB) to find attractants. R. pseudosolanacearum D8 showed strong responses to green pepper and carrot juices and TSB. We constructed a mutant library of R. pseudosolanacearum D8 by deleting each of the MCP genes. Chemotaxis assays to TSB revealed that an MCP which we named McpD was responsible for sensing an attractant(s) in TSB. Because amino acids are the major constituents of TSB, we measured chemotactic responses of R. pseudosolanacearum D8 to 20 proteinogenic amino acids and found Asp and Glu as the major attractants of McpD and Cys as the minor attractant. R. pseudosolanacearum Ps29 can utilize Asp and Glu as sole carbon and nitrogen sources, suggesting that the role of McpD-mediated chemotaxis is finding growth substrates.","PeriodicalId":15842,"journal":{"name":"Journal of General and Applied Microbiology","volume":" ","pages":""},"PeriodicalIF":0.8,"publicationDate":"2025-07-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143639722","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

Screening to isolate Bacillus subtilis mutants with enhanced NADPH levels. 提高NADPH水平的枯草芽孢杆菌突变体的筛选。

IF 0.8 4区生物学

Journal of General and Applied Microbiology Pub Date : 2025-07-15 Epub Date: 2025-01-16 DOI: 10.2323/jgam.2025.01.001

Yuzheng Wu, Shu Ishikawa, Ken-Ichi Yoshida

引用次数: 0

Identification of Escherichia coli flagellar antigen by disc immuno-immobilization. 圆盘免疫固定化法鉴定大肠杆菌鞭毛抗原。

IF 0.8 4区生物学

Journal of General and Applied Microbiology Pub Date : 2025-07-15 Epub Date: 2025-04-23 DOI: 10.2323/jgam.2025.04.001

Shiho Oyama, Masatoshi Fujihara

引用次数: 0

Production of p-anisaldehyde via whole-cell transformation using recombinant E. coli expressing trans-anethole oxygenase. 表达反式茴香酚加氧酶的重组大肠杆菌全细胞转化生产对茴香醛。

IF 0.8 4区生物学

Journal of General and Applied Microbiology Pub Date : 2025-07-15 Epub Date: 2025-02-18 DOI: 10.2323/jgam.2025.02.001

Zhikai Zhang, Qian Lin

{"title":"Production of p-anisaldehyde via whole-cell transformation using recombinant E. coli expressing trans-anethole oxygenase.","authors":"Zhikai Zhang, Qian Lin","doi":"10.2323/jgam.2025.02.001","DOIUrl":"10.2323/jgam.2025.02.001","url":null,"abstract":"p-Anisaldehyde, a fragrance and flavour with important roles in food, cosmetics, and drug industries, is currently synthesized through chemical methods. Production of p-anisaldehyde by chemical oxidation of trans-anethole in industry gives rise to excessive by-products and adverse environmental impacts, whereas biological process would address such problems. Here, we presented a process of biotransformation of trans-anethole for production of p-anisaldehyde. The tao gene encoding for trans-anethole oxygenase (TAO) from Paraburkholderia sp. MR185 was fused with a solubilization tag GST and ProS2, respectively. GST did not exhibit solubility enhancement effect, whereas fusion with ProS2 significantly improved TAO's soluble expression in E. coli and the fusion protein ProS2-TAO-Sil3K accounted for more than 40% of total soluble proteins. ProS2-TAO-Sil3K was purified by simple silica affinity and its activity did not require addition of NADH, NADPH, and FAD. Metal ions Co2+, Zn2+, Ni2+, and Cu2+ displayed significant inhibition effect on TAO activity, and addition of Fe2+ improved enzyme activity by 32.6%. After induction, engineered E. coli cells were used as whole-cell biocatalyst for transformation of trans-anethole, and the final concentration of p-anisaldehyde reached 10.18 mM (1.38 g/L), with the volumetric productivity of 0.11 g/L/h and conversion rate of 67.9%. These results reveal that the biosynthesis of p-anisaldehyde has a great potential in practice.","PeriodicalId":15842,"journal":{"name":"Journal of General and Applied Microbiology","volume":" ","pages":""},"PeriodicalIF":0.8,"publicationDate":"2025-07-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143458238","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

Screening system for MAA-CoA productivity using 2-methylcitrate biosensor. 2-甲基柠檬酸盐生物传感器MAA-CoA产率筛选系统。

IF 0.8 4区生物学

Journal of General and Applied Microbiology Pub Date : 2025-06-20 DOI: 10.2323/jgam.2025.05.003

Satoshi Hasegawa, Ryota Kato, Daichi Ishihara, Miyu Tsukada, Takumi Ojima, Shigeko Kawai-Noma, Daisuke Umeno

引用次数: 0

Functional characterization of the mys genes for porphyra-334 biosynthesis from the terrestrial cyanobacterium Nostoc commune by heterologous expression. 陆生蓝藻Nostoc commune生物合成卟啉-334基因的异源表达功能表征

IF 0.8 4区生物学

Journal of General and Applied Microbiology Pub Date : 2025-05-30 DOI: 10.2323/jgam.2025.05.002

Wei Yang, Toshio Sakamoto

{"title":"Functional characterization of the mys genes for porphyra-334 biosynthesis from the terrestrial cyanobacterium Nostoc commune by heterologous expression.","authors":"Wei Yang, Toshio Sakamoto","doi":"10.2323/jgam.2025.05.002","DOIUrl":"https://doi.org/10.2323/jgam.2025.05.002","url":null,"abstract":"Mycosporine-like amino acids (MAAs) are low-molecular-weight UV-protective compounds, and porphyra-334 and shinorine are common MAAs. Porphyra-334 is synthesized via the conjugation of mycosporine-glycine with threonine, whereas substitution with serine yields shinorine. The terrestrial cyanobacterium Nostoc commune KU002 (NIES-2538) produces 7-O-(β-arabinopyranosyl)-porphyra-334, and the mysABCD gene cluster responsible for MAA biosynthesis has been isolated. The heterologous expression of the mysABC genes from N. commune KU002 in Escherichia coli cells led to mycosporine-glycine production regardless of the culture medium supplemented with serine, threonine, or xylose. When the mysABCD genes from N. commune KU002 were expressed in E. coli cells, porphyra-334 production occurred, and shinorine production was observed upon serine supplementation in the culture medium. Notably, threonine and xylose supplementation in the culture medium increased the amounts of porphyra-334 in both cellular extracts and culture medium extracts. When the mysD gene was replaced with that from the shinorine producer Actinosynnema mirum JCM 3225, shinorine was primarily synthesized instead of porphyra-334. Interestingly, the transformant expressing the chimeric KU002-mysABC-JCM3225-mysD produced a novel MAA derivative with an absorption maximum at 334 nm and a molecular mass of 346 when cultured in the medium supplemented with threonine and xylose. These results suggest that the substrate specificity of MysD, which catalyzes the conjugation of mycosporine-glycine and serine or threonine, alters the production of porphyra-334 or shinorine and that the supplements added to the culture medium affect the amount and composition of MAAs produced in the E. coli transformant.","PeriodicalId":15842,"journal":{"name":"Journal of General and Applied Microbiology","volume":" ","pages":""},"PeriodicalIF":0.8,"publicationDate":"2025-05-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144199345","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

Secretory Expression of a Multifunctional Nuclease Nuc A in Bacillus licheniformis 2709. 多功能核酸酶Nuc a在地衣芽孢杆菌2709中的分泌表达。

IF 0.8 4区生物学

Journal of General and Applied Microbiology Pub Date : 2025-05-29 DOI: 10.2323/jgam.2025.05.001

Huimin Guo, Kefen Wang, Tongtong Zhang, Honglei Fang, Wei Hui, Huitu Zhang

{"title":"Secretory Expression of a Multifunctional Nuclease Nuc A in Bacillus licheniformis 2709.","authors":"Huimin Guo, Kefen Wang, Tongtong Zhang, Honglei Fang, Wei Hui, Huitu Zhang","doi":"10.2323/jgam.2025.05.001","DOIUrl":"https://doi.org/10.2323/jgam.2025.05.001","url":null,"abstract":"Serratia nuclease Nuc A is a non-specific nucleotide hydrolase that has been widely used in large-scale protein purification or eliminating nucleic acid contamination from purified proteins. To enhance the enzyme production, the Serratia nuclease gene was synthesized and expressed in Bacillus licheniformis 2709, a robust strain capable of secreting native and heterologous proteins selectively or non-selectively. To further increase the secretory expression level of the enzyme, different strong promoters and signal peptides were fused with the mature Nuc A-encoding gene at various genetic loci. The highest expression level of Nuc A was observed under the control of regulatory elements PaprE, which occur naturally in B. licheniformis 2709 for the alkaline protease (AprE) expression. Through maximizing the number of copies of PaprE-nucA expression cassette at different integration sites, the yield of nuclease Nuc A reached approximately 31954 U/mL after 60 hours of cultivation in shake flasks. The specific activity of the recombinant nuclease reached 1.58×107 U/mg, which is about 9 times higher than that expressed in Escherichia coli strain. Additionally, the recombinant Nuc A exhibited high catalytic activities in the pH range of 7-10. Furthermore, it was resistant to 0.2% SDS, 1.0 mM PMSF, and 0.4% Triton X-100. After 8 M Urea treatment, residual activity is measured. The high expression levels and positive characteristics of recombinant Nuc A provide an effective solution for large-scale production and industrial application of the nuclease.","PeriodicalId":15842,"journal":{"name":"Journal of General and Applied Microbiology","volume":" ","pages":""},"PeriodicalIF":0.8,"publicationDate":"2025-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144174138","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