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Lactic acid bacteria from Ethiopian traditional beverage, Tella: technological and metabolic profiles for industrial application: Gashaw Assefa Yehuala, Jaein Choe, Nurelegne Tefera Shibeshi, Kumsa Delessa, Asnake Desalegn, Mi-Kyung Park; J. Microbiol. 2025;63(1):e.2409008. Published online December 20, 2024; DOI: https://doi.org/10.71150/jm.2409008

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Tella is a traditional beverage widely accepted by consumers, despite the lack of product consistency owing to its reliance on natural fermentation. This study aimed to identify potential industrial lactic acid bacteria (LAB) starter cultures based on their technological properties. Seven LAB strains isolated from Tella were characterized for their carbohydrate utilization, salt content, temperature, and acid tolerances, growth and acidification rates, and metabolite profiles. Most strains efficiently utilized various carbohydrates, with Lactiplantibacillus plantarum TDM41 showing exceptional versatility. The strains exhibited similar growth characteristics. Principal component analysis of stress tolerance properties revealed that L. plantarum TDM41, Pediococcus pentosaceus TAA01, and Leuconostoc mesenteroides TDB22 exhibited superior tolerance ability. Strong acidification properties were detected in the L. plantarum TDM41, P. pentosaceus TAA01, and Leuconostoc mesenteroides TDB22 strains after 24 h incubation at 30°C. L. plantarum TDM41 displayed the fastest acidification rate throughout the analysis period. All LAB strains produced significant amounts of diverse organic acids, including lactic acid, citric acid, acetic acid, malic acid, and succinic acid, with lactic acid being the primary acid produced by each strain. Overall, strains L. plantarum TDM41 and P. pentosaceus TAA01 prove to be potential candidates for Tella industrial starter cultures and similar cereal products owing to their robust technological properties.

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Preparation method and physicochemical characteristics of Tella: an Ethiopian fermented beverage
Rabira Lemessa Gudeta, Solomon Abera, Hirpha Adugna Areti
Journal of Ethnic Foods.2025;[Epub] CrossRef

Review

Reverse Zoonotic Transmission of SARS-CoV-2 and Monkeypox Virus: A Comprehensive Review: Chiranjib Chakraborty, Manojit Bhattacharya, Md Aminul Islam, Hatem Zayed, Elijah Ige Ohimain, Sang-Soo Lee, Prosun Bhattacharya, Kuldeep Dhama; J. Microbiol. 2024;62(5):337-354. Published online May 23, 2024; DOI: https://doi.org/10.1007/s12275-024-00138-9

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Reverse zoonosis reveals the process of transmission of a pathogen through the human-animal interface and the spillback of the zoonotic pathogen. In this article, we methodically demonstrate various aspects of reverse zoonosis, with a comprehensive discussion of SARS-CoV-2 and MPXV reverse zoonosis. First, different components of reverse zoonosis, such as humans, different pathogens, and numerous animals (poultry, livestock, pets, wild animals, and zoo animals), have been demonstrated. Second, it explains the present status of reverse zoonosis with different pathogens during previous occurrences of various outbreaks, epidemics, and pandemics. Here, we present 25 examples from literature. Third, using several examples, we comprehensively illustrate the present status of the reverse zoonosis of SARS-CoV-2 and MPXV. Here, we have provided 17 examples of SARS-CoV-2 reverse zoonosis and two examples of MPXV reverse zoonosis. Fourth, we have described two significant aspects of reverse zoonosis: understanding the fundamental aspects of spillback and awareness. These two aspects are required to prevent reverse zoonosis from the current infection with two significant viruses. Finally, the One Health approach was discussed vividly, where we urge scientists from different areas to work collaboratively to solve the issue of reverse zoonosis.

Citations

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Phylogenetic analyses of the spread of Clade I MPOX in African and non-African nations
Chiranjib Chakraborty, Manojit Bhattacharya, Arpita Das, Ali S. Abdelhameed
Virus Genes.2025; 61(3): 265. CrossRef
Efficient and modular reverse genetics system for rapid generation of recombinant severe acute respiratory syndrome coronavirus 2
Sojung Bae, Jinjong Myoung
Journal of Microbiology.2025; 63(7): e2504015. CrossRef
Real-time malaria detection in the Amazon rainforest via drone-collected eDNA and portable qPCR
Yin Cheong Aden Ip, Luca Montemartini, Jia Jin Marc Chang, Andrea Desiderato, Nicolás D. Franco-Sierra, Christian Geckeler, Mailyn Adriana Gonzalez Herrera, Michele Gregorini, Meret Jucker, Steffen Kirchgeorg, Martina Lüthi, Elvira Mächler, Frederik Bendi
One Health.2025; 21: 101167. CrossRef
One digital health through wearables: a viewpoint on human–pet integration towards Healthcare 5.0
Mostafa Haghi, Samira Abani, Soheil Khooyooz, Anice Jahanjoo, Samaneh Rashidibajgan, Nima TaheriNejad, Thomas M. Deserno, Holger Volk
Frontiers in Digital Health.2025;[Epub] CrossRef
Development of a multiplex real-time PCR for the simultaneous detection of monkeypox virus clades I, II, and goatpox virus
Yongqiang Lin, Zijing Guo, Jinsong Chen, Xianwen Zhang, Long Zhou, Yanmin Li, Zhidong Zhang
Frontiers in Veterinary Science.2024;[Epub] CrossRef
Differential Impact of Spike Protein Mutations on SARS-CoV-2 Infectivity and Immune Evasion: Insights from Delta and Kappa Variants
Tae-Hun Kim, Sojung Bae, Jinjong Myoung
Journal of Microbiology and Biotechnology.2024; 34(12): 2506. CrossRef

Journal Articles

Chemokine CCL6 Plays Key Role in the Inhibitory Effect of Vitamin A on Norovirus Infection: Heetae Lee , Giljae Lee , You-Hee Cho , Youngcheon Song , GwangPyo Ko; J. Microbiol. 2023;61(5):579-587. Published online May 26, 2023; DOI: https://doi.org/10.1007/s12275-023-00047-3

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Abstract PDF: Norovirus (NoV) is the most common viral cause of acute gastroenteritis worldwide. Vitamin A has demonstrated the potential to protect against gastrointestinal infections. However, the effects of vitamin A on human norovirus (HuNoV) infections remain poorly understood. This study aimed to investigate how vitamin A administration affects NoV replication. We demonstrated that treatment with retinol or retinoic acid (RA) inhibited NoV replication in vitro based on their effects on HuNoV replicon-bearing cells and murine norovirus-1 (MNV-1) replication in murine cells. MNV replication in vitro showed significant transcriptomic changes, which were partially reversed by retinol treatment. RNAi knockdown of CCL6, a chemokine gene that was downregulated by MNV infection but upregulated by retinol administration, resulted in increased MNV replication in vitro. This suggested a role of CCL6 in the host response to MNV infections. Similar gene expression patterns were observed in the murine intestine after oral administration of RA and/or MNV-1.CW1. CCL6 directly decreased HuNoV replication in HG23 cells, and might indirectly regulate the immune response against NoV infection. Finally, relative replication levels of MNV-1.CW1 and MNV-1.CR6 were significantly increased in CCL6 knockout RAW 264.7 cells. This study is the first to comprehensively profile transcriptomes in response to NoV infection and vitamin A treatment in vitro, and thus may provide new insights into dietary prophylaxis and NoV infections.

Cytophaga hutchinsonii chu_2177, encoding the O-antigen ligase, is essential for cellulose degradation: Yahong Tan , Wenxia Song , Lijuan Gao , Weican Zhang , Xuemei Lu; J. Microbiol. 2022;60(4):364-374. Published online January 7, 2022; DOI: https://doi.org/10.1007/s12275-022-1531-3

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Abstract PDF

Cytophaga hutchinsonii can efficiently degrade crystalline cellulose, in which the cell surface cellulases secreted by the type IX secretion system (T9SS) play important roles, but the degradation mechanism remains unclear, and the anchor mechanism of cellulases on the outer membrane in C. hutchinsonii has not been studied. Here, chu_2177 was identified by transposon mutagenesis and was proved to be indispensable for cellulose utilization in C. hutchinsonii. Disruption of chu_2177 resulted in O-antigen deficiency and chu_ 177 could confer O-antigen ligase activity upon an Escherichia coli waal mutant, indicating that chu_2177 encoded the Ontigen ligase. Moreover, deletion of chu_2177 caused defects in cellulose utilization, cell motility, biofilm formation, and stress resistance. Further study showed that the endoglucanase activity was markedly decreased in the outer membrane but was increased in the culture fluid without chu_2177. Western blot proved that endoglucanase CHU_1336 was not located on the outer membrane but was released in the culture fluid of the Δ2177 mutant. Further proteomics analysis showed that many cargo proteins of T9SS were missing in the outer membrane of the Δ2177 mutant. Our study revealed that the deletion of chu_2177 affected the localization of many T9SS cargo proteins including cellulases on the outer membrane of C. hutchinsonii.

Citations

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Screening and genome-wide analysis of lignocellulose-degrading bacteria from humic soil
Tianjiao Zhang, Shuli Wei, Yajie Liu, Chao Cheng, Jie Ma, Linfang Yue, Yanrong Gao, Yuchen Cheng, Yongfeng Ren, Shaofeng Su, Xiaoqing Zhao, Zhanyuan Lu
Frontiers in Microbiology.2023;[Epub] CrossRef
The type IX secretion system: Insights into its function and connection to glycosylation in Cytophaga hutchinsonii
Wenxia Song, Xueke Zhuang, Yahong Tan, Qingsheng Qi, Xuemei Lu
Engineering Microbiology.2022; 2(3): 100038. CrossRef

Improved tolerance of Escherichia coli to oxidative stress by expressing putative response regulator homologs from Antarctic bacteria: Seo-jeong Park , Sangyong Lim , Jong-il Choi; J. Microbiol. 2020;58(2):131-141. Published online December 23, 2019; DOI: https://doi.org/10.1007/s12275-020-9290-5

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Response regulator (RR) is known a protein that mediates cell’s response to environmental changes. The effect of RR from extremophiles was still under investigation. In this study, response regulator homologs were mined from NGS data of Antarctic bacteria and overexpressed in Escherichia coli. Sixteen amino acid sequences were annotated corresponding to response regulators related to the two-component regulatory systems; of these, 3 amino acid sequences (DRH632, DRH1601 and DRH577) with high homology were selected. These genes were cloned in pRadGro and expressed in E. coli. The transformant strains were subjected to various abiotic stresses including oxidative, osmotic, thermal stress, and acidic stress. There was found that the robustness of E. coli to abiotic stress was increased in the presence of these response regulator homologs. Especially, recombinant E. coli overexpressing drh632 had the highest survival rate in oxidative, hypothermic, osmotic, and acidic conditions. Recombinant E. coli overexpressing drh1601 showed the highest tolerance level to osmotic stress. These results will be applicable for development of recombinant strains with high tolerance to abiotic stress.

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Mechanistic and bibliometric insights into RpoS -mediated biofilm regulation and its strategic role in food safety applications
Shirin Akter, Md. Ashikur Rahman, Md. Ashrafudoulla, A.G.M.Sofi Uddin Mahamud, Md Anamul Hasan Chowdhury, Sang-Do Ha
Critical Reviews in Food Science and Nutrition.2025; 65(30): 7070. CrossRef
Adaption strategies of extremophiles and the construction of wastewater treatment systems driven by extremophiles
Zheng Guo, Yong-Guang Li, Zhi-Bin Wang, Xin Zhou, Shou-Qing Ni
Environmental Research.2025; 282: 121979. CrossRef
Deionococcus proteotlycius Genomic Library Exploration Enhances Oxidative Stress Resistance and Poly-3-hydroxybutyrate Production in Recombinant Escherichia coli
Seul-Ki Yang, Soyoung Jeong, Inwoo Baek, Jong-il Choi, Sangyong Lim, Jong-Hyun Jung
Microorganisms.2023; 11(9): 2135. CrossRef
Bacterial redox response factors in the management of environmental oxidative stress
Sudharsan M, Rajendra Prasad N, Saravanan Rajendrasozhan
World Journal of Microbiology and Biotechnology.2023;[Epub] CrossRef
Bacteriophages as Antimicrobial Agents? Proteomic Insights on Three Novel Lytic Bacteriophages Infecting ESBL-Producing Escherichia coli
Sadika Dkhili, Miguel Ribeiro, Salma Ghariani, Houssem Ben Yahia, Mélanie Hillion, Patricia Poeta, Karim Ben Slama, Michel Hébraud, Gilberto Igrejas
OMICS: A Journal of Integrative Biology.2021; 25(10): 626. CrossRef
Regulator of ribonuclease activity modulates the pathogenicity of Vibrio vulnificus
Jaejin Lee, Eunkyoung Shin, Jaeyeong Park, Minho Lee, Kangseok Lee
Journal of Microbiology.2021; 59(12): 1133. CrossRef

Sterilization efficiency of pathogen-contaminated cottons in a laundry machine: Yoonjae Shin , Jungha Park , Woojun Park; J. Microbiol. 2020;58(1):30-38. Published online November 25, 2019; DOI: https://doi.org/10.1007/s12275-020-9391-1

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Abstract PDF

Pathogenic bacteria on abiotic surfaces such as fabrics, bedding, patient wears, and surgical tools are known to increase the risk of bacterial diseases in infants and the elderly. The desiccation tolerance of bacteria affects their viability in cotton. Thus, washing and drying machines are required to use conditions that ensure the sterilization of bacteria in cotton. The objective of this study is to determine the effects of various sterilization conditions of washing and drying machines on the survival of three pathogenic bacteria (Acinetobacter baumannii, Pseudomonas aeruginosa, and Staphylococcus aureus) commonly presented in contaminated cotton and two non-pathogenic bacteria (Bacillus subtilis and Escherichia coli) in cotton. High survival rates of A. baumannii and S. aureus in desiccated cotton were observed based on scanning electron microscope and replicate organism direct agar contact assay. The survival rates of A. baumannii and S. aureus exposed in desiccated cotton for 8 h were higher (14.4 and 5.0%, respectively) than those of other bacteria (< 0.5%). All tested bacteria were eradicated at low-temperature (< 40°C) washing with activated oxygen bleach (AOB). However, bacterial viability was shown in low temperature washing without AOB. High-temperature (> 60°C) washing was required to achieve 99.9% of the sterilization rate in washing without AOB. The sterilization rate was 93.2% using a drying machine at 60°C for 4 h. This level of sterilization was insufficient in terms of time and energy efficiency. High sterilization efficiency (> 99.9%) at 75°C for 3 h using a drying machine was confirmed. This study suggests standard conditions of drying machines to remove bacterial contamination in cotton by providing practical data.

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Reviews

MINIREVIEW] Bacterial persistence: Fundamentals and clinical importance: Sung-Hee Jung , Choong-Min Ryu , Jun-Seob Kim; J. Microbiol. 2019;57(10):829-835. Published online August 28, 2019; DOI: https://doi.org/10.1007/s12275-019-9218-0

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Abstract PDF

The threat of antibiotic-resistant bacteria is increasing worldwide. Bacteria utilize persistence and resistance to survive antibiotic stress. For a long time, persistence has been studied only under laboratory conditions. Hence, studies of bacterial persistence are limited. Recently, however, the high incidence of infection relapses caused by persister cells in immunocompromised patients has emphasized the importance of persister research. Furthermore, persister pathogens are one of the causes of chronic infectious diseases, leading to the overuse of antibiotics and the emergence of antibiotic-resistant bacteria. Therefore, understanding the precise mechanism of persister formation is important for continued use of available antibiotics. In this review, we aimed to provide an overview of the persister studies published to date and the current knowledge of persister formation mechanisms. Recent studies of the features and mechanisms of persister formation are analyzed from the perspective of the nature of the persister cell.

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REVIEW] Ribosome dependence of persister cell formation and resuscitation: Thomas K. Wood , Sooyeon Song , Ryota Yamasaki; J. Microbiol. 2019;57(3):213-219. Published online February 26, 2019; DOI: https://doi.org/10.1007/s12275-019-8629-2

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Abstract PDF

Since most bacterial cells are starving, they must enter a resting stage. Persister is the term used for metabolically-dormant cells that are not spores, and these cells arise from stress such as that from antibiotics as well as that from starvation. Because of their lack of metabolism, persister cells survive exposure to multiple stresses without undergoing genetic change; i.e., they have no inherited phenotype and behave as wild-type cells once the stress is removed and nutrients are presented. In contrast, mutations allow resistant bacteria to grow in the presence of antibiotics and slow growth allows tolerant cells to withstand higher concentrations of antibiotics; hence, there are three closely-related phenotypes: persistent, resistant, and tolerant. In addition, since dormancy is so prevalent, persister cells must have a means for resuscitating (since so many cells should obtain this resting state). In this review, we focus on what is known about the formation and resuscitation of persister cells.

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Journal Articles

Overexpression and characterization of a novel cold-adapted and salt-tolerant GH1 β-glucosidase from the marine bacterium Alteromonas sp. L82: Jingjing Sun , Wei Wang , Congyu Yao , Fangqun Dai , Xiangjie Zhu , Junzhong Liu , Jianhua Hao; J. Microbiol. 2018;56(9):656-664. Published online August 23, 2018; DOI: https://doi.org/10.1007/s12275-018-8018-2

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Abstract PDF

A novel gene (bgl) encoding a cold-adapted β-glucosidase was cloned from the marine bacterium Alteromonas sp. L82. Based on sequence analysis and its putative catalytic conserved region, Bgl belonged to the glycoside hydrolase family 1. Bgl was overexpressed in E. coli and purified by Ni2+ affinity chromatography. The purified recombinant β- glucosidase showed maximum activity at temperatures between 25°C to 45°C and over the pH range 6 to 8. The enzyme lost activity quickly after incubation at 40°C. Therefore, recombinant β-glucosidase appears to be a cold-adapted enzyme. The addition of reducing agent doubled its activity and 2 M NaCl did not influence its activity. Recombinant β-glucosidase was also tolerant of 700 mM glucose and some organic solvents. Bgl had a Km of 0.55 mM, a Vmax of 83.6 U/mg, a kcat of 74.3 s-1 and kcat/Km of 135.1 at 40°C, pH 7 with 4-nitrophenyl-β-D-glucopyranoside as a substrate. These properties indicate Bgl may be an interesting candidate for biotechnological and industrial applications.

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Comparison of antibiotic resistance and copper tolerance of Enterococcus spp. and Lactobacillus spp. isolated from piglets before and after weaning: Xueting Zou , Mengwei Weng , Xu Ji , Rong Guo , Weijiang Zheng , Wen Yao; J. Microbiol. 2017;55(9):703-710. Published online September 2, 2017; DOI: https://doi.org/10.1007/s12275-017-6241-x

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In China, antimicrobials and copper are used extensively as growth-promoting agents for piglets. This study aimed to characterize the role of in-feed copper in the emergence of copper-tolerant and antibiotic-resistant Enterococcus and Lactobacillus isolates in Chinese pig farms. Feces of the same eight piglets from four litters at 7 and 55 days old and their mothers were traced in order to isolate Enterococcus spp. and Lactobacillus spp.. The minimum inhibitory concentrations of 10 antimicrobials and copper sulfate were determined using an agar dilution method. The feed levels of Cu2+ for lactating sows, suckling piglets, and weaned piglets were 6, 177, and 18 mg/kg, respectively. All the 136 Enterococcus isolates were sensitive to vancomycin; and the resistance rates to penicillin, enrofloxacin, and high level streptomycin resistance increased significantly after weaning. For the 155 Lactobacillus isolates, the resistance rates to ampicillin, chloramphenicol, tetracycline, and enrofloxacin were significantly higher in weaned piglets. The ratios of copper tolerant Enterococcus and Lactobacillus isolates both increased significantly after weaning (P < 0.05). A phenotypic correlation was observed after classifying the isolates into two groups (CuSO4 MIC50 < 16 or 􎃶􀁇16 for enterococci; CuSO4 MIC50 < 12 or 􎃶􀁇12 for lactobacilli) and comparing the antimicrobial-resistant percentage of two groups. On species level, a significant increase of E. faecalis to enrofloxacin was observed in line with the increase of copper MIC (P < 0.05). The findings revealed the changes of the antibiotic resistance and copper tolerance level of enterococci and lactobacilli between suckling and weaned piglets and demonstrated that there might be a strong association between in-feed copper and increased antibiotic resistance in enterococci and lactobacilli in Chinese intensive swine farms.

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Research Support, Non-U.S. Gov'ts

The effects of fluctuating culture temperature on stress tolerance and antioxidase expression in Esteya vermicola: Yun-bo Wang , Wen-xing Pang , Xiao-na Yv , Jing-jie Li , Yong-an Zhang , Chang-keun Sung; J. Microbiol. 2015;53(2):122-126. Published online January 28, 2015; DOI: https://doi.org/10.1007/s12275-015-4529-2

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The endoparasitic nematophagous fungus, Esteya vermicola, has shown great potential as a biological control agent against the pine wood nematode, Bursaphelenchus xylophilus. Fluctuating culture temperatures can affect fungal yields and fungal tolerance to desiccation, UV radiation, H2O2, and heat stress, as well as antioxidase expression. To explore these effects, E. vermicola cultured under five temperature ranges, 26oC, 15-26oC, 26-35oC, 20-30oC, and 15-35oC, were compared. The cultures grown at lower temperatures showed better growth, stronger tolerance to desiccation, UV, and H2O2 stresses, and increased catalase expression, However, these cultures also showed weaker heat stress tolerance and lower superoxide dismutase expression than the higher-temperature cultures. In particular, the E. vermicola cultured at 20-30oC, i.e., fluctuating in a narrow range around the optimal temperature, showed the best performance. Therefore, for production in practical applications, this narrowly fluctuating, moderate temperature appears to be optimal for yield and stress tolerance in E. vermicola.

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Screening and Optimization of Solid-State Fermentation for Esteya vermicola, an Entomopathogenic Fungus Against the Major Forest Pest Pine Wood Nematode
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In vivo infection of Bursaphelenchus xylophilus by the fungus Esteya vermicola
Hai‐Hua Wang, Yun‐Bo Wang, Can Yin, Jie Gao, Ran Tao, Yu‐Lou Sun, Chun‐Yan Wang, Zhen Wang, Yong‐Xia Li, Chang‐Keun Sung
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Detection of Inhibitors of Phenotypically Drug-tolerant Mycobacterium tuberculosis Using an In Vitro Bactericidal Screen: Ian M. Bassett , Shichun Lun , William R. Bishai , Haidan Guo , Joanna R. Kirman , Mudassar Altaf , Ronan F. O’Toole; J. Microbiol. 2013;51(5):651-658. Published online June 25, 2013; DOI: https://doi.org/10.1007/s12275-013-3099-4

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Many whole cell screens of chemical libraries currently in use are based on inhibition of bacterial growth. The goal of this study was to develop a chemical library screening model that enabled detection of compounds that are active against drug-tolerant non-growing cultures of Mycobacterium tuberculosis. An in vitro model of low metabolically active mycobacteria was established with 8 and 30 day old cultures of M. smegmatis and M. tuberculosis, respectively. Reduction of resazurin was used as a measure of viability and the assay was applied in screens of chemical libraries for bactericidal compounds. The model provided cells that were phenotypically-resilient to killing by first and second-line clinical drugs including rifampicin. Screening against chemical libraries identified proteasome inhibitors, NSC310551 and NSC321206, and a structurally-related series of thiosemicarbazones, as having potent killing activity towards aged cultures. The inhibitors were confirmed as active against virulent M. tuberculosis strains including multi- and extensively-drug resistant clinical isolates. Our library screen enabled detection of compounds with a potent level of bactericidal activity towards phenotypically drug-tolerant cultures of M. tuberculosis.

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Role of Heavy Metal Resistant Ochrobactrum sp. and Bacillus spp. Strains in Bioremediation of a Rice Cultivar and Their PGPR Like Activities: Sanjeev Pandey , Pallab Kumar Ghosh , Sisir Ghosh , Tarun Kumar De , Tushar Kanti Maiti; J. Microbiol. 2013;51(1):11-17. Published online March 2, 2013; DOI: https://doi.org/10.1007/s12275-013-2330-7

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The present study demonstrates the metal toxicity ameliorating and growth promoting abilities of three different bacterial isolates when applied to rice as host plant. The three bacterial strains included a cadmium resistant Ochrobactrum sp., a lead resistant Bacillus sp. and an arsenic resistant Bacillus sp. designated as CdSP9, PbSP6, and AsSP9, respectively. When these isolates were used as inocula applied to metaltreated rice plants of variety Satabdi, the germination percentage, relative root elongation (RRE), amylase and protease activities were increased. The toxic effect of metal was reduced in presence of these bacteria. The overall biomass and root/shoot ratio were also enhanced by bacterial inoculation. Hydroponic studies showed that the superoxide dismutase (SOD) activity and malondialdehyde (MDA) level, which had been increased in the presence of metal stress in rice roots, were lowered by the bacterial inoculation. In addition, all three strains were 1-aminocyclopropane-1-carboxylate (ACC) deaminase and catalase positive, whereas siderophore producing ability was lacking in PbSP6. However, both PbSP6 and AsSP9 were protease positive and could hydrolyse starch. The data indicate that these bacteria have promise for bioremediation as well as for plant growth promotion.

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Enhancement of Butanol Tolerance and Butanol Yield in Clostridium acetobutylicum Mutant NT642 Obtained by Nitrogen Ion Beam Implantation: Xiao-Bo Liu , Qiu-Ya Gu , Xiao-Bin Yu , Wei Luo; J. Microbiol. 2012;50(6):1024-1028. Published online December 30, 2012; DOI: https://doi.org/10.1007/s12275-012-2289-9

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Abstract PDF

As a promising alternative biofuel, biobutanol can be produced through acetone/butanol/ethanol (ABE) fermentation. Currently, ABE fermentation is still a small-scale industry due to its low production and high input cost. Moreover, butanol toxicity to the Clostridium fermentation host limits the accumulation of butanol in the fermentation broth. The wild-type Clostridium acetobutylicum D64 can only produce about 13 g butanol/L and tolerates less than 2% (v/v) butanol. To improve the tolerance of C. acetobutylicum D64 for enhancing the production of butanol, nitrogen ion beam implantation was employed and finally five mutants with enhanced butanol tolerance were obtained. Among these, the most butanol tolerant mutant C. acetobutylicum NT642 can tolerate above 3% (v/v) butanol while the wide-type strain can only withstand 2% (v/v). In batch fermentation, the production of butanol and ABE yield of C. acetobutylicum NT642 was 15.4 g/L and 22.3 g/L, respectively, which were both higher than those of its parental strain and the other mutants using corn or cassava as substrate. Enhancing butanol tolerance is a great precondition for obtaining a hyperyield producer. Nitrogen ion beam implantation could be a promising biotechnology to improve butanol tolerance and production of the host strain C. acetobutylicum.

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NOTE] Identification of Chaperones in Freeze Tolerance in Saccharomyces cerevisiae: Mahendran Chinnamara Naicker , I Seul Jo , Hana Im; J. Microbiol. 2012;50(5):882-887. Published online November 4, 2012; DOI: https://doi.org/10.1007/s12275-012-2411-z

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Abstract: Exposure to low temperatures reduces protein folding rates and induces the cold denaturation of proteins. Considering the roles played by chaperones in facilitating protein folding and preventing protein aggregation, chaperones must exist that confer tolerance to cold stress. Here, yeast strains lacking individual chaperones were screened for reduced freezing tolerance. In total, 19 of 82 chaperone-deleted strains tested were more sensitive to freeze-thaw treatment than wild-type cells. The reintroduction of the respective chaperone genes into the deletion mutants recovered the freeze tolerance. The freeze sensitivity of the chaperone-knockout strains was also retained in the presence of 20% glycerol.

Journal Article

Screening for Probiotic Properties of Strains Isolated from Feces of Various Human Groups: Sathyaseelan Sathyabama , Rajendran Vijayabharathi , Palanisamy Bruntha devi , Manohar Ranjith kumar , Venkatesan Brindha Priyadarisini; J. Microbiol. 2012;50(4):603-612. Published online July 21, 2012; DOI: https://doi.org/10.1007/s12275-012-2045-1

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The present study searched for potential probiotic strains from various human fecal samples. A total of 67 aerobic and 38 anaerobic strains were isolated from 5 different categories of human feces. Systematic procedures were used to evaluate the probiotic properties of the isolated strains. These showed about 75–97% survivability in acidic and bile salt environments. Adhesion to intestinal cell line Caco-2 was also high. The isolates exhibited hydrophobic properties in hexadecane. The culture supernatants of these strains showed antagonistic effects against pathogens. The isolates were resistant to a simulated gastrointestinal environment in vitro. Of the 4 best isolates, MAbB4 (Staphylococcus succinus) and FIdM3 (Enterococcus fecium), were promising candidates for a potential probiotic. S. succinus was found to be a probiotic strain, which is the second such species reported to date in this particular genus. A substantial zone of inhibition was found against Salmonella spp., which adds further support to the suggestion that the probiotic strain could help prevent intestinal infection. This study suggested that the human flora itself is a potential source of probiotics.

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Research Support, Non-U.S. Gov'ts

The ATPase Activity of The G2alt Gene Encoding an Aluminium Tolerance Protein from Anoxybacillus gonensis G2: Fatih Saban Beris , Lina De Smet , Hakan Karaoglu , Sabriye Canakci , Jozef Van Beeumen , Ali Osman Belduz; J. Microbiol. 2011;49(4):641-650. Published online September 2, 2011; DOI: https://doi.org/10.1007/s12275-011-0522-6

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Abstract PDF

The G2ALT gene was cloned and sequenced from the thermophilic bacterium Anoxybacillus gonensis G2. The gene is 666 bp long and encodes a protein 221 amino acids in length. The gene was overexpressed in E. coli and purified to homogeneity and biochemically characterized. The enzyme has a molecular mass of 24.5 kDa and it could be classified as a member of the family of bacterial aluminium resistance proteins based on homology searches. When this fragment was expressed in E. coli, it endowed E. coli with Al tolerance to 500 μM. The purified G2ALT protein is active at a broad pH range (pH 4.0-10.0) and temperature range (25°C-80°C) with optima of 6.0 and the apparent optimal temperature of 73°C respectively. Under optimal conditions, G2ALT exhibited a low ATPase activity with Km- and Vmax- values of 10±0.55 μM and 26.81±0.13 mg Pi released/min/mg enzyme, respectively. The ATPase activity of G2ALT requires Mg2+ and Na+ ions, while Zn2+ and Al3+ stimulate the activity. Cd2+ and Ag+ reduced the activity and Li+, Cu2+, and Co2+ inhibited the activity. Known inhibitors of most ATPases, like such as β-mercaptoethanol and ouabain, also inhibited the activity of the G2ALT. These biochemical characterizations suggested that G2ALT belongs to the PP-loop ATPase superfamily and it can be responsible for aluminium tolerance in A. gonensis G2.

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Cel8H, a Novel Endoglucanase from the Halophilic Bacterium Halomonas sp. S66-4: Molecular Cloning, Heterogonous Expression, and Biochemical Characterization: Xiaoluo Huang , Zongze Shao , Yuzhi Hong , Ling Lin , Chanjuan Li , Fei Huang , Hui Wang , Ziduo Liu; J. Microbiol. 2010;48(3):318-324. Published online June 23, 2010; DOI: https://doi.org/10.1007/s12275-009-0188-5

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Abstract PDF

A recombinant Escherichia coli clone expressing an endoglucanase was identified from a genomic library of the halophilic bacterium Halomonas sp. S66-4, and the enzyme was designated Cel8H. The cel8H gene consisted of 1,053 bp and encoded 350 amino acids sharing the highest identity of 48% to other known endoglucanases. The protein was expressed in E. coli BL21 (DE3) and purified to homogeneity. The purified recombinant enzyme had an optimal activity of 4.9 U/mg at pH 5 and 45°C toward the substrate carboxymethylcellulose. It exhibited extraordinary properties which differed from endoglucanases reported previously at the point of high salt tolerance above 5 M, simultaneously with high pH stability at pH 4-12 and high temperature stability at 40-60°C. Various substrate tests indicated that the enzyme hydrolyzes β-1,4-glucosidic bonds specifically.

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Isolation, Characterization, and Evaluation of Wild Isolates of Lactobacillus reuteri from Pig Feces: Deog Yong Lee , Yeon-Soo Seo , Nabin Rayamajhi , Mi Lan Kang , Su In Lee , Han Sang Yoo; J. Microbiol. 2009;47(6):663-672. Published online February 4, 2010; DOI: https://doi.org/10.1007/s12275-009-0124-8

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Abstract PDF

Lactic acid bacteria (LAB) are a well-used probiotics for health improvements in both humans and animals. Despite of several benefits, non-host-specific LAB showed poor probiotics effects due to difficulty in colonization and competition with normal flora. Therefore, the feasibility of porcine LAB isolates was evaluated as a probiotics. Ten of 49 Lactobacillus spp. isolates harbored 2~10 kb plasmid DNA. Seven strains were selected based on the safety test, such as hemolytic activity, ammonia, indole, and phenylalanine production. After safety test, five strains were selected again by several tests, such as epithelial adherence, antimicrobial activity, tolerance against acid, bile, heat, and cold-drying, and production of acid and hydrogen peroxide. Then, enzyme profiles (ZYM test) and antibiotics resistance were analyzed for further characterization. Five Lactobacillus reuteri isolates from pig feces were selected by safety and functional tests. The plasmid DNA which was able to develop vector system was detected in the isolates. Together with these approaches, pig-specific Lactobacillus spp. originated from pigs were selected. These strains may be useful tools to develop oral delivery system.

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Dark Septate Endophyte (DSE) Fungi Isolated from Metal Polluted Soils: Their Taxonomic Position, Tolerance, and Accumulation of Heavy Metals In Vitro: Yujie Zhang , Yan Zhang , Maojun Liu , Xiaodong Shi , Zhiwei Zhao; J. Microbiol. 2008;46(6):624-632. Published online December 24, 2008; DOI: https://doi.org/10.1007/s12275-008-0163-6

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Abstract PDF

To understand the possible role of the plant root associated fungi on metal tolerance, their role in the uptake of heavy metals and the potential transfer of these metal ions to the plant, three strains of dark septate endophytic (DSE) fungi were isolated from a waste smelter site in southwest China, and one strain was isolated from a non-contaminated site. According to molecular phylogenetic analysis of the ITS 1-5.8S rDNA-ITS 2 gene regions and morphological characteristics, one is identified as Exophiala pisciphila, and the other three are non-sporulating fungi under the experiment condition with the nearest phylogenetic affinities to the Thysanorea papuana strain EU041814. Tolerance and accumulation abilities of the three DSE strains for metals were investigated in liquid culture. Minimum inhibitory concentrations (MIC) of Pb, Zn, and Cd were determined. It was demonstrated that the tolerance of the DSE strains varied between metal species and strains. The E. pisciphila strain is able to accumulate lead and cadmium over 20% and 5% of dry weight of biomass, respectively. Partial of the sequestrated metals can be washed with CaCl2. Morphological and enzyme activity changes taking place in the presence of excessive Pb, Cd, and/or Zn also indicate that the mechanism of heavy metal tolerance and accumulation of the DSE strains would be a complex process. The findings indicated promising tolerance and accumulation of the DSE strains with potential values in metal cycling and restoration of soil and water system.

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Diversity and Metal Tolerance of Nematode-Trapping Fungi in Pb-Polluted Soils: Ming-He Mo , Wei-Min Chen , Hao-Ran Yang , Ke-Qin Zhang; J. Microbiol. 2008;46(1):16-22.; DOI: https://doi.org/10.1007/s12275-007-0174-8

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Abstract PDF

The diversity of nematode-trapping fungi (NTF) in two lead (Pb) mines in Yunnan Province, China was investigated in 2004. In total, 20 species belonging to five genera were identified from 500 samples collected at the Lanping and the Huize mines. Pb concentrations ranged from 216~7,150 mg/kg for the former and 132~13,380 mg/kg for the latter, respectively. The fungi were divided into five groups based on different trapping mechanisms. The trapping-net producer group contained the largest number of species, with nine. Two predators, Dactylellina ellipsosporum and Arthrobotrys oligospora, were found at frequencies of 32.85% and 15.41%, respectively. The diversity indexes of NTF were positively correlated with Pb pollution levels in both the Lanping Mine (r=0.66) and the Huize Mine (r=0.72), suggesting that the distribution of NTF was not negatively affected by Pb contamination. For most strains of a given species, there was no significant difference (P>0.01) in the Pb tolerance between the strains isolated from habitats with low or high Pb concentrations. However, Pb toxicity exerted adverse effects on trap formation and predacious capability of fungi. We discuss the possible metal tolerance mechanisms and their relationships to the survival strategy of NTF in Pb-polluted environments.

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Induction of ethanol tolerance on the production of 17-ketosteroids by mutant of mycobacterium sp.: Kim, Mal Nam , Kim, Eun Mi; J. Microbiol. 1995;33(4):322-327.

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Abstract PDF: Tolerance of Mycobacterium sp. against organic solvents has been induced for the cholesterol side chain degradation by adding chemicals associated with synthesis of fatty acids or alcohols. Biotin of 300 ㎍/1 and 0.5% aqueous ethanol solution were optima for the enhancement of ethanol tolerance of the microorganism. The induction of ethanol tolerance by biotin was found to be due to increase of degree of unsaturation of the fatty acids in membranous phospholipid of the cell, especially due to increase of oleic acid content. However when 0.5% of ethanol was added for the ethanol tolerance induction, there was an ambiguous correlation between ethanol tolerance and degree of unsaturation of the fatty acids, in spite of the fact that the induction increased the content of unsaturated fatty acids. Addition of 0.5% of ethanol induced several ethanol shock proteins having molecular weight similar to that of heat shock proteins.

Identification of the Genes Involved in Stationary-Phase Specific Acid Resistance of Salmonella typhimurium: Bang, Iel Soo , Lee, In Soo , Lee, Yung Nok , Park, Yong keun; J. Microbiol. 1995;33(1):21-27.

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Abstract PDF: Salmonella encounters variables pH fluctuation during its life cycle and has been developed adaptative systems such as acid tolerance response (ATR) to survive at severe acidic environment. As part of on going investigation of stationary-phase specific acid resistance, we have searched for acid sensitive mutations in virulent Salmonella typhimurim UK-1 usin the MudJ fusion technique and two lethal selection procedures including DNP(dinitrophenol) selection media and microtiterplate selection method. Two acid sensitive mutations have been identified and designated, spatrK2, spatrK5. These mutations removed both stationary-phase acid tolerant effect and stationary-phase specific acid resistance. Non-specific histone like protein, H-NS and stationary-phase specific sigma factor, RpoS made little contribution to that system at respective single mutation(5-10 fold decrease). But, when both mutations were combined together, no acid resistance was achieved while acid tolerance response was still effective. Two dimensional SDS polyacrylamide gel electrophoresis showed new stationary-phase specific acid shock proteins as well as proteins already known. Not expectedly, the gels from acid adapted samples of both rpoS and hns mutation showed that double mutation of those regulators does not make change of the standard acid shock proteins. Only four acid shock proteins were regulated by these regulators, while fifteen proteins were newly identified as the members of acid shock response system by these regulators. These results implicate that stationary-phase acid resistance of that organism has RpoS/H-NS soubly dependent acid protective system and independent acid tolerance response system.

Osmotic Tolerance Response of Salmonella typhimurium with Respect ro Growth-Phase and Identification of otr201, a rpoS-Related Gene: Lim, Si Keun , Bang, Soo Iel , Bang, Seong Ho , Lee, Yung Nok , Park, Yong Keun; J. Microbiol. 1995;33(1):66-73.

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Abstract PDF: Salmonella typhimurium can stand against and survive under lethal osmotic exposure. Two systems of osmotic tolerance response(OTR) were found to be utilized by that organism, which were possibly overlapping with each other. The first system is an induction in response to non-lethal high osmoshock(0.3~0.7 M NaCl) at log-phase. The second system is induced during famine condition of stationary-phase. The viability of wild types(UK1, LT2) under these unfavorable conditions was increased by both systems. The viability of stationary-phase cells was approximately 5-fold that of the cells adapted at log-phase. In addition, a few regulatory fenes(rpoS, fur, crp, atp), one carbonstarvation-inducible(cstA104), and an osmotic-inducible gene(proU) were found to play an important role in osmotic tolerance at both growth phases. RpoS, a putative alternative sigma factor (σ^38), was found to participate in OTR systems regardless of growth-phase, but rpoS-defective mutant could still develope the adaptive tolerance. Thus, we concluded that there is rpoS-defective and rpoS-independent systems for osmotic tolerance at both growth-phase. Of the possible otr mutants newly isolated using MudJ(Km, lac) operon fusion techniques, YK3092 (otr201::MudJ) was most sensitive to osmotic challenge regardless of growth phase. It was mapped nearby at 57 min on chromosome and showed rpoS-negative phenotypes such as no catalase activity and inability to accumulate glycogen : but was not linked to rpoS. Therefore, this result strongly suggest that otr201 might be a rpoS-related regulatory gene not gound before.

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