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Review
Expanding the genetic code: In vivo approaches for incorporating non-proteinogenic monomers
Dongheon Lee, Suk Min Yun, Jong-il Choi
J. Microbiol. 2025;63(3):e2501005.   Published online March 28, 2025
DOI: https://doi.org/10.71150/jm.2501005
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AbstractAbstract PDF

The application of genetic code expansion has enabled the incorporation of non-canonical amino acids (ncAAs) into proteins, introducing novel functional groups and significantly broadening the scope of protein engineering. Over the past decade, this approach has extended beyond ncAAs to include non-proteinogenic monomers (npMs), such as β-amino acids and hydroxy acids. In vivo incorporation of these monomers requires maintaining orthogonality between endogenous and engineered aminoacyl-tRNA synthetase (aaRS)/tRNA pairs while optimizing the use of the translational machinery. This review introduces the fundamental principles of genetic code expansion and highlights the development of orthogonal aaRS/tRNA pairs and ribosomal engineering to incorporate npMs. Despite these advancements, challenges remain in engineering aaRS/tRNA pairs to accommodate npMs, especially monomers that differ significantly from L-α-amino acids due to their incompatibility with existing translational machinery. This review also introduces recent methodologies that allow aaRSs to recognize and aminoacylate npMs without reliance on the ribosomal translation system, thereby unlocking new possibilities for synthesizing biopolymers with chemically diverse monomers.

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  • Advancing microbial engineering through synthetic biology
    Ki Jun Jeong
    Journal of Microbiology.2025; 63(3): e2503100.     CrossRef
Journal Article
Hydroxychloroquine an Antimalarial Drug, Exhibits Potent Antifungal Efficacy Against Candida albicans Through Multitargeting
Sargun Tushar Basrani, Tanjila Chandsaheb Gavandi, Shivani Balasaheb Patil, Nandkumar Subhash Kadam, Dhairyasheel Vasantrao Yadav, Sayali Ashok Chougule, Sankunny Mohan Karuppayil, Ashwini Khanderao Jadhav
J. Microbiol. 2024;62(5):381-391.   Published online April 8, 2024
DOI: https://doi.org/10.1007/s12275-024-00111-6
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AbstractAbstract
Candida albicans is the primary etiological agent associated with candidiasis in humans. Unrestricted growth of C. albicans can progress to systemic infections in the worst situation. This study investigates the antifungal activity of Hydroxychloroquine (HCQ) and mode of action against C. albicans. HCQ inhibited the planktonic growth and yeast to hyphal form morphogenesis of C. albicans significantly at 0.5 mg/ml concentration. The minimum inhibitory concentrations (MIC(50)) of HCQ for C. albicans adhesion and biofilm formation on the polystyrene surface was at 2 mg/ml and 4 mg/ml respectively. Various methods, such as scanning electron microscopy, exploration of the ergosterol biosynthesis pathway, cell cycle analysis, and assessment of S oxygen species (ROS) generation, were employed to investigate HCQ exerting its antifungal effects. HCQ was observed to reduce ergosterol levels in the cell membranes of C. albicans in a dose-dependent manner. Furthermore, HCQ treatment caused a substantial arrest of the C. albicans cell cycle at the G0/G1 phase, which impeded normal cell growth. Gene expression analysis revealed upregulation of SOD2, SOD1, and CAT1 genes after HCQ treatment, while genes like HWP1, RAS1, TEC1, and CDC 35 were downregulated. The study also assessed the in vivo efficacy of HCQ in a mice model, revealing a reduction in the pathogenicity of C. albicans after HCQ treatment. These results indicate that HCQ holds for the development of novel antifungal therapies.
Review
The “Cins” of Our Fathers: Rejuvenated Interest in Colicins to Combat Drug Resistance
Sumudu Upatissa , Robert J. Mitchell
J. Microbiol. 2023;61(2):145-158.   Published online February 8, 2023
DOI: https://doi.org/10.1007/s12275-023-00023-x
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AbstractAbstract
With the growing threat of antibiotic resistance, researchers around the globe are seeking alternatives to stem bacterial pathogenesis. One such alternative is bacteriocins, proteins produced by bacterial species to inhibit the growth and viability of related bacterial species. With their diverse mechanisms, which include pore formation and nuclease activities, and narrow spectrum of activities, which limit their impact to only certain bacterial species, unlike many chemical antibiotics, bacteriocins offer intriguing possibilities to selectively control individual bacterial populations. Within this review, therefore, we highlight current research exploring the application of colicins and microcins, a subset of bacteriocins, with an emphasis on their activities against drug-resistant pathogens, both in in vitro and in vivo settings.

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  • Isolation, Genomics-Based and Biochemical Characterization of Bacteriocinogenic Bacteria and Their Bacteriocins, Sourced from the Gastrointestinal Tract of Meat-Producing Pigs
    Ester Sevillano, Irene Lafuente, Nuria Peña, Luis M. Cintas, Estefanía Muñoz-Atienza, Pablo E. Hernández, Juan Borrero
    International Journal of Molecular Sciences.2024; 25(22): 12210.     CrossRef
  • Intelligent Biological Networks: Improving Anti-Microbial Resistance Resilience through Nutritional Interventions to Understand Protozoal Gut Infections
    Avinash V. Karpe, David J. Beale, Cuong D. Tran
    Microorganisms.2023; 11(7): 1800.     CrossRef
  • Pairing Colicins B and E5 with Bdellovibrio bacteriovorus To Eradicate Carbapenem- and Colistin-Resistant Strains of Escherichia coli
    Sumudu Upatissa, Wonsik Mun, Robert J. Mitchell, Minsu Kim
    Microbiology Spectrum.2023;[Epub]     CrossRef
  • Bacteriocin-Producing Escherichia coli Q5 and C41 with Potential Probiotic Properties: In Silico, In Vitro, and In Vivo Studies
    Veronika S. Mihailovskaya, Dmitry A. Sutormin, Marina O. Karipova, Anna B. Trofimova, Victor A. Mamontov, Konstantin Severinov, Marina V. Kuznetsova
    International Journal of Molecular Sciences.2023; 24(16): 12636.     CrossRef
Journal Article
Paenibacillus lycopersici sp. nov. and Paenibacillus rhizovicinus sp. nov., isolated from the rhizosphere of tomato (Solanum lycopersicum)
Shin Ae Lee , Tae-Wan Kim , Mee-Kyung Sang , Jaekyeong Song , Soon-Wo Kwon , Hang-Yeon Weon
J. Microbiol. 2020;58(10):832-840.   Published online September 29, 2020
DOI: https://doi.org/10.1007/s12275-020-0258-2
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AbstractAbstract
Two Gram-stain-positive, rod-shaped, endospore-forming bacteria, designated 12200R-189T and 14171R-81T were isolated from the rhizosphere of tomato plants. The 16S rRNA gene sequence similarity between strains 12200R-189T and 14171R-81T were 97.2%. Both strains showed the highest 16S rRNA gene sequence similarities to Paenibacillus sacheonensis SY01T (96.3% and 98.0%, respectively). The genome of strain 12200R-189T was approximately 6.7 Mb in size with 5,750 protein-coding genes (CDSs) and the G + C content was 58.1 mol%, whereas that of strain 14171R-81T comprised one chromosome of 7.0 Mb and two plasmids (0.2 Mb each) with 6,595 CDSs and the G + C content was 54.5 mol%. Comparative genome analysis revealed that average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values among 12200R-189T, 14171R-81T, and other closely related species were below the cut-off levels 95% and 70%, respectively. Strain 12200R-189T grew at a temperature range of 15–40°C, pH 6.0–9.0, and 0–3% NaCl (w/v), whereas strain 14171R-81T grew at a temperature range of 10–37°C, pH 6.0– 8.0, and 0–1% NaCl (w/v). Menaquinone-7 (MK-7) was the only isoprenoid quinone detected in both strains. The predominant cellular fatty acids (> 10%) were iso-C15:0, anteiso- C15:0, and iso-C16:0. The polar lipids of strain 12200R- 189T were diphosphatidylglycerol (DPG), phosphatidylglycerol (PG), phosphatidylethanolamine (PE), aminophospholipid (APL), phospholipid (PL), phosphatidylglycolipid (PGL), and four aminophosphoglycolipids (APGLs) and those of strain 14171R-81T were DPG, PG, PE, APL, three PLs, two PGLs, and three APGLs. Based on phylogenetic, genomic, phenotypic, and chemotaxonomic analyses, strains 12200R- 189T and 14171R-81T represent two novel species of the genus Paenibacillus, for which the names Paenibacillus lycopersici sp. nov. and Paenibacillus rhizovicinus sp. nov. are proposed. The type strains are 12200R-189T (= KACC 19916T = CCTCC AB 2020027T) and 14171R-81T (= KACC 19915T = CCTCC AB 2020026T).

Citations

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  • Description and genomic characterization of Jiella flava sp. nov., isolated from Acrostichum aureum
    Ming-Sheng Chen, Xiu-Long Pu, Ming-Dan Weng, Li Chen, Lan-Ying Zhu, Li Tuo
    International Journal of Systematic and Evolutionary Microbiology .2022;[Epub]     CrossRef
  • Jiella sonneratiae sp. nov., a novel endophytic bacterium isolated from bark of Sonneratia apetala
    Ming-Sheng Chen, Hai-Bo Yi, Zi-Hao Huang, Xiao-Rui Yan, Xiao-Hui Chen, Xiao Ma, Zhou-Qing Zheng, Li Tuo
    International Journal of Systematic and Evolutionary Microbiology .2022;[Epub]     CrossRef
  • Paenibacillus vietnamensis sp. nov., isolated from the rhizosphere soil of Arachis hypogaea
    Minh Hong Nguyen, Mai Thi Ngoc Dinh, Keun Chul Lee, Ji-Sun Kim, Thao Kim Nu Nguyen, Jung-Sook Lee
    International Journal of Systematic and Evolutionary Microbiology .2022;[Epub]     CrossRef
  • Effect of exopolysaccharides of Paenibacillus polymyxa rhizobacteria on physiological and morphological variables of wheat seedlings
    Irina V. Yegorenkova, Kristina V. Tregubova, Alexander I. Krasov, Nina V. Evseeva, Larisa Yu. Matora
    Journal of Microbiology.2021; 59(8): 729.     CrossRef
Physiological Relevance of Salt Environment for in vitro recA System
Kim , Jong Il
J. Microbiol. 1999;37(2):59-65.
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AbstractAbstract
RecA protein can promote strand assimilation, homologous pairing, and strand exchange. All these reactions require DNA-dependent ATP hydrolysis by recA protein, and the activities of recA protein are affected by the ionic environment. In this experiment, DNA-dependent ATPase activity showed different sensitivity to anionic species. ATP hydrolysis and strand exchange were relatively sensitive to salt in the reactions with NaCl, strongly inhibited at 100 mM NaCl. However, the inhibition by sodium acetate or sodium glutamate was not observed at 50∼100 mM concentration. Addition of sodium glutamate to the standard reaction condition increased the apparent efficiency of ATP hydrolysis during strand exchange. The condition including 50∼100 mM sodium-glutamate might be similar to the physiological condition.

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