Journal of Microbiology

Journal Articles

Ultrasonic Treatment Enhanced Astaxanthin Production of Haematococcus pluvialis: Yun Hwan Park , Jaewon Park , Jeong Sik Choi , Hyun Soo Kim , Jong Soon Choi , Yoon-E Choi; J. Microbiol. 2023;61(6):633-639. Published online June 13, 2023; DOI: https://doi.org/10.1007/s12275-023-00053-5

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In this study, effects of ultrasonic treatment on Haematococcus pluvialis (H. pluvialis) were investigated. It has been confirmed that the ultrasonic stimulation acted as stress resources in the red cyst stage H. pluvialis cells containing astaxanthin,
result
ing in additional astaxanthin production. With the increase in production of astaxanthin, the average diameter of H. pluvialis cells increased accordingly. In addition, to determine how ultrasonic stimulation had an effect on the further biosynthesis of astaxanthin, genes related to astaxanthin synthesis and cellular ROS level were measured. As a result, it was confirmed that astaxanthin biosynthesis related genes and cellular ROS levels were increased, and thus ultrasonic stimulation acts as an oxidative stimulus. These results support the notion on the effect of the ultrasonic treatment, and we believe our novel approach based on the ultrasonic treatment would help to enhance the astaxanthin production from H. pluvialis.

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Recent Advances in Astaxanthin as an Antioxidant in Food Applications
Yimeng Dang, Zhixi Li, Fanqianhui Yu
Antioxidants.2024; 13(7): 879. CrossRef
Effect of reduced atmospheric pressures on the morphology and astaxanthin biosynthesis of microalga Haematococcus lacustris
Sangui Kim, Rendi Mahadi, Aditya Lakshmi Narasimhan, Catherine Christabel, Hyoji Yu, Eui-Jin Kim, You-Kwan Oh
Biotechnology and Bioprocess Engineering.2024; 29(6): 1131. CrossRef

Metformin Regulates Gut Microbiota Abundance to Suppress M2 Skewing of Macrophages and Colorectal Tumorigenesis in Mice: Linfeng Fan , Xiangfu Zeng , Guofeng Xu; J. Microbiol. 2023;61(1):109-120. Published online January 26, 2023; DOI: https://doi.org/10.1007/s12275-022-00010-8

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Abstract

The correlation of imbalanced gut microbiota with the onset and progression of colorectal cancer (CRC) has become clear. This work investigates the effect of metformin on gut microbiota and genesis of CRC in mice. Human fecal samples were collected from healthy control (HC) donors and CRC patients. Compared to HC donors, CRC patients had reduced abundance of gut microbiota; however, they had increased abundance of detrimental Bacteroidetes. Mice were injected with azomethane (AOM) to induce colorectal tumorigenesis models. Treatment of CRC patients-sourced fecal microbiota promoted tumorigenesis, and it increased the expression of Ki67, β-catenin, COX-2, and Cyclin D1 in mouse colon tissues. Further treatment of metformin blocked the colorectal tumorigenesis in mice. Fecal microbiota from the metformin-treated mice was collected, which showed decreased Bacteroidetes abundance and suppressed AOM-induced colorectal tumorigenesis in mice as well. Moreover, the metformin- modified microbiota promoted the M1 macrophage-related markers IL-6 and iNOS but suppressed the M2 macrophage-related markers IL-4R and Arg1 in mouse colon tissues. In conclusion, this study suggests that metformin-mediated gut microbiota alteration suppresses macrophage M2 polarization to block colorectal tumorigenesis.

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Metformin alleviates colitis-associated colorectal cancer via inhibition of the TLR4/MyD88/NFκB/MAPK pathway and macrophage M2 polarization
Xueying Lai, Bin Liu, Yu Wan, Ping Zhou, Wanjun Li, Wei Hu, Wei Gong
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Wenting Li, Nanshu Liu, Mingwei Chen, Dongjuan Liu, Sai Liu
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Linxin Zhou, Linfu Zheng, Binbin Xu, Zhou Ye, Dazhou Li, Wen Wang
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An AMPK agonist suppresses the progress of colorectal cancer by regulating the polarization of TAM to M1 through inhibition of HIF-1α and mTOR signal pathway
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Journal of Cancer Research and Therapeutics.2023; 19(6): 1560. CrossRef

Effect of biostimulation and bioaugmentation on hydrocarbon degradation and detoxification of diesel-contaminated soil: a microcosm study: Patricia Giovanella , Lídia de Azevedo Duarte , Daniela Mayumi Kita , Valéria Maia de Oliveira , Lara Durães Sette; J. Microbiol. 2021;59(7):634-643. Published online May 15, 2021; DOI: https://doi.org/10.1007/s12275-021-0395-2

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Abstract

Soil contamination with diesel oil is quite common during processes of transport and storage. Bioremediation is considered a safe, economical, and environmentally friendly approach for contaminated soil treatment. In this context, studies using hydrocarbon bioremediation have focused on total petroleum hydrocarbon (TPH) analysis to assess process effectiveness, while ecotoxicity has been neglected. Thus, this study aimed to select a microbial consortium capable of detoxifying diesel oil and apply this consortium to the bioremediation of soil contaminated with this environmental pollutant through different bioremediation approaches. Gas chromatography (GC-FID) was used to analyze diesel oil degradation, while ecotoxicological bioassays with the bioindicators Artemia sp., Aliivibrio fischeri (Microtox), and Cucumis sativus were used to assess detoxification. After 90 days of bioremediation, we found that the biostimulation and biostimulation/ bioaugmentation approaches showed higher rates of diesel oil degradation in relation to natural attenuation (41.9 and 26.7%, respectively). Phytotoxicity increased in the biostimulation and biostimulation/bioaugmentation treatments during the degradation process, whereas in the Microtox test, the toxicity was the same in these treatments as that in the natural attenuation treatment. In both the phytotoxicity and Microtox tests, bioaugmentation treatment showed lower toxicity. However, compared with natural attenuation, this approach did not show satisfactory hydrocarbon degradation. Based on the microcosm experiments results, we conclude that a broader analysis of the success of bioremediation requires the performance of toxicity bioassays.

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

Involvement of Alternative Oxidase in the Regulation of Sensitivity of Sclerotinia sclerotiorum to the Fungicides Azoxystrobin and Procymidone: Ting Xu , Ya-Ting Wang , Wu-Sheng Liang , Fei Yao , Yong-Hong Li , Dian-Rong Li , Hao Wang , Zheng-Yi Wang; J. Microbiol. 2013;51(3):352-358. Published online April 26, 2013; DOI: https://doi.org/10.1007/s12275-013-2534-x

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Abstract

Sclerotinia sclerotiorum is a filamentous fungal pathogen that can infect many economically important crops and vegetables. Alternative oxidase is the terminal oxidase of the alternative respiratory pathway in fungal mitochondria. The function of alternative oxidase was investigated in the regulation of sensitivity of S. sclerotiorum to two commercial fungicides, azoxystrobin and procymidone which have different fungitoxic mechanisms. Two isolates of S. sclerotiorum were sensitive to both fungicides. Application of salicylhydroxamic acid, a specific inhibitor of alternative oxidase, significantly increased the values of effective concentration causing 50% mycelial growth inhibition (EC50) of azoxystrobin to both S. sclerotiorum isolates, whereas notably decreased the EC50 values of procymidone. In mycelial respiration assay azoxystrobin displayed immediate inhibitory effect on cytochrome pathway capacity, but had no immediate effect on alternative pathway capacity. In contrast, procymidone showed no immediate impact on capacities of both cytochrome and alternative pathways in the mycelia. However, alternative oxidase encoding gene (aox) transcript and protein levels, alternative respiration pathway capacity of the mycelia were obviously increased by pre-treatment for 24 h with both azoxystrobin and procymidone. These results indicate that alternative oxidase was involved in the regulation of sensitivity of S. sclerotiorum to the fungicides azoxystrobin and procymidone, and that both fungicides could affect aox gene expression and the alternative respiration pathway capacity development in mycelia of this fungal pathogen.

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Involvement of Alternative Oxidase in the Regulation of Growth, Development, and Resistance to Oxidative Stress of Sclerotinia sclerotiorum: Ting Xu , Fei Yao , Wu-Sheng Liang , Yong-Hong Li , Dian-Rong Li , Hao Wang , Zheng-Yi Wang; J. Microbiol. 2012;50(4):594-602. Published online August 25, 2012; DOI: https://doi.org/10.1007/s12275-012-2015-7

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Abstract

Sclerotinia sclerotiorum is a cosmopolitan, filamentous, fungal pathogen that can cause serious disease in many kinds of crops. Alternative oxidase is the terminal oxidase of the alternative mitochondrial respiratory pathway in fungi and higher plants. We report the presence of this alternative pathway respiration and demonstrate its expression in two isolates of S. sclerotiorum under unstressed, normal culture conditions. Application of salicylhydroxamic acid, a specific inhibitor of alternative oxidase, severely inhibited the mycelial growth of S. sclerotiorum both on potato dextrose agar plates and in liquid culture media. Inhibition of alternative oxidase could influence the growth pattern of S. sclerotiorum, as salicylhydroxamic acid treatment induced obvious aerial mycelia growing on potato dextrose agar plates. Under the treatment with salicylhydroxamic acid, S. sclerotiorum formed sclerotia much more slowly than the control. Treatment with hydrogen peroxide in millimolar concentrations greatly decreased the growth rate of mycelia and delayed the formation of sclerotia in both tested S. sclerotiorum isolates. As well, this treatment obviously increased their alternative pathway respiration and the levels of both mRNA and protein of the alternative oxidase. These results indicate that alternative oxidase is involved in the regulation of growth, development, and resistance to oxidative stress of S. sclerotiorum.

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Differential Expression of citA Gene Encoding the Mitochondrial Citrate Synthase of Aspergillus nidulans in Response to Developmental Status and Carbon Sources: In Sook Min , Ji Young Bang , Soon Won Seo , Cheong Ho Lee , Pil Jae Maeng; J. Microbiol. 2010;48(2):188-198. Published online May 1, 2010; DOI: https://doi.org/10.1007/s12275-010-0096-8

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Abstract

As an extension of our previous studies on the mitochondrial citrate synthase of Aspergillus nidulans and cloning of its coding gene (citA), we analyzed differential expression of citA in response to the progress of development and change of carbon source. The cDNA consisted of 1,700 nucleotides and was predicted to encode a 474-amino acid protein. By comparing the cDNA sequence with the corresponding genomic sequence, we confirmed that citA gene contains 7 introns and that its transcription starts at position -26 (26-nucleotide upstream from the initiation codon). Four putative CreA binding motifs and three putative stress-response elements (STREs) were found within the 1.45-kb citA promoter region. The mode of citA expression was examined by both Northern blot and confocal microscopy using green fluorescent protein (sGFP) as a vital reporter. During vegetative growth and asexual development, the expression of citA was ubiqiutous throughout the whole fungal body including mycelia and conidiophores. During sexual development, the expression of citA was quite strong in cleistothecial shells, but significantly weak in the content of cleistothecia including ascospores. Acetate showed a strong inductive effect on citA expression, which is subjected to carbon catabolite repression (CCR) caused by glucose. The recombinant fusion protein CitA40::sGFP (sGFP containing the 40-amino acid N-terminal segment of CitA) was localized into mitochondria, which supports that a mitochondrial targeting signal is included within the 40-amino acid N-terminal segment of CitA.

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The Role and Regulation of Trx1, a Cytosolic Thioredoxin in Schizosaccharomyces pombe: Ji-Yoon Song , Jung-Hye Roe; J. Microbiol. 2008;46(4):408-414. Published online August 31, 2008; DOI: https://doi.org/10.1007/s12275-008-0076-4

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Abstract

The genome of fission yeast Schizosaccharomyces pombe harbors two genes for thioredoxins, trx1+ and trx2+, which encode cytosolic and mitochondrial thioredoxins, respectively. The Δtrx1 mutant was found sensitive to diverse external stressors such as various oxidants, heat, and salt, whereas Δtrx2 mutant was not sensitive except to paraquat, a superoxide generator. Both Δtrx1 and Δtrx2 mutants were more resistant to diamide, a thiol-specific oxidant, than the wild type. The trx1+ gene expression was induced by H2O2 and menadione, being mediated through a stress-responsive transcription factor Pap1. In Δtrx1 cells, the basal expression of Pap1-regulated genes were elevated, suggesting a role for Trx1 as a reducer for oxidized (activated) Pap1. The Δtrx1 mutant exhibited cysteine auxotrophy, which can be overcome by adding sulfite. This suggests that Trx1 serves as a primary electron donor for 3’-phosphoadenosine-5’-phosphosulfate (PAPS) reductase and thus is an essential protein for sulfur assimilation in S. pombe. These results suggest that, in contrast to Trx2 whose role is more confined to mitochondrial functions, Trx1 plays a major role in protecting S. pombe against various stressful conditions and enables proper sulfur metabolism.

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Restriction and Transcription Maps of Mitochondrial DNA of Trimorphomyces papilionaceus: Jeoung, Won Jin , Hong, Soon Gyu , Kang, Young Won , Jung, Hack Sung; J. Microbiol. 1995;33(2):149-153.

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Abstract: Mitochondrial DNA has been isolated from Trimorphomyces papilionaceus. By analyzing DNA fragments digested by restriction enzymes, a restriction site map has been constructured. The mtDNA of T. papilionaceus amounts to 48.5 kb in size and is circular in structure. Entire mitochondrial DNA was cloned in E coli plasmids and Northern blot hybridization was done using cloned and subcloned DNAs as probes. Based on hybridization results of mitochondrial RNA transcripts, a transcription map was prepared.

Phylogenetic study of trichaptum species based on the RFLP analysis of mitochondrial DNA: Kim, Mi Sun , jung, Hack Sung; J. Microbiol. 1996;34(3):215-219.

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Abstract: Eight strains of Trichaptunm (Polyporaceae), two strains from each species of T. abietinum, T. biforme, T. fusco-violaceum, and T. laricinum were examined to see their phylogenetic relationship by digesting mitochondrial DNAs with EcoRV, Hind III, XbaI, and PstI, and then analyzing fragmentation patterns with the methods of Nei and Li. T. abietinum, T. biforme, and T. laricinum developed an independent phylogenetic lineage, respectively, but T. fusco-violaceum FP-133997-sp showed a close relationship with two strains of T. bioforme, and T. fusco-violaceum HHB-4016-sp barely grouped with those of T. laricinum. Based on the results of the RFLP analysis of mtDNA, it is concluded that T. fusco-violaceum is under way to differentiation into two different subgroups.

Amino acid substitutions conferring cold-sensitive phenotype on the yeast MTF1 gene: Jang , Sei Heon; J. Microbiol. 1997;35(3):228-233.

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Abstract: The MTF1 gene of Saccharomyces cerevisiae encodes a 43 kDa MITOCHONDRIAL RNA polymerase specificity factor which recognizes mitochondrial promoters to initiate correct transcription. To better understand structure-function of the MTF1 gene as well as the transcription mechanism of mitochondrial RNA polymerase, two cold-sensitive alleles of the MTF1 mutation were isolated by plasmid shuffling method after PCR-based random mutagenesis of the MTF1 gene. The mutation sites were analyzed by nucleotide sequencing. These cs phenotype mtf1 mutants were respiration competent on the nonfermentible glycerol medium at the permissive temperature, but incompetent at 13℃. The cs phenotype allele of the MTF1, yJH147, encoded an L146P replacement. The other cs allele, yJH148, contained K179E and K214M double replacements. Mutations in both alleles were in a region of Mtflp which is located between domains with amino acid sequence similarities to conserved regions 2 and 3 of bacterial s factors.

The Genetic Organization of the Linear Mitochondrial Plasmid mlp1 from Pleurotus ostreatus NFFA2: Kim, Eun Kyung , Youn, Hye Sook , Koo, Yong Bom , Roem Jung Hye; J. Microbiol. 1997;35(4):264-270.

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Abstract: The structure of plasmid mlp1, a linear 10.2kb mitochondrial plasmid of Pleurotus ostreatus NFF A2 was determined by restriction enzyme mapping and partial sequencing. The plasmid encodes at least two proteins; a putative RNA polymerase showing homology to yeast mitochondrial RNA polymerase and to viral-encoded RNA polymerases, and a putative DNA polymerase showing significant homology to the family B thpe DNA polymerases. It also contains terminal inverted repeat sequences at both ends which are longer than 274 bp. A 1.6 kb EcoRI restriction fragment of m1p1 containing the putative RNA polymerase gene did not hybridize to the nuclear or motochondrial genomes from P. ostreatus, suggesting that it may encode plasmidspecific RNA polymerase. The gene fragment also did not hybridize with the RNA polymerase gene (RPO41) from Saccaromyces cerevisiae. The relationship between genes in m1p1 and those in another linear plasmid pC1K1 of Claviceps purpurea was examined by DNA hybridization. The result indicates that the genes for DNA and RNA polymerases are not closely related with those in C. purpurea.

Identification of Critical Amino Acids in the Core RNA Polynerase Binding Region of Yease Mtflp: Yang, Jae Sub , Jang, Sei Heon; J. Microbiol. 1998;36(3):208-213.

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Abstract: Yeast mitochondral RNA polymerase specificity factor encoded by the nuclear MTF1 gene is required for a selective transcription on nonanucleotide mitochondral promoter by core RNA polymerase. Although there is a little amino acid sequence similarity of Mtf1p with bacterial sigma factors, the mode of transcriptional initiation of mitochondrial RNA polymerase is identical to that of E. coli RNA polymerase. To study the interaction of mtf1p with core polymerase, we carried out region-directed random mutagenesis of the core binding domain with the pool of mutant oligonucleotide. Out of 4,000 transformants screened for petite phenotype on glycerol media by plasmid shuffling, six alleles of the MTF1 gene were isolated. The positions of amino acid replacements that resulted in mtf1 mutants were limited to amino acids 53-54 and 65-67. Among mutant forms of Mtf1p overproduced in E. coli, Mtf1p with either L53H or Y65Dmutation was unable to produce a selective transcript in run-off transcription reaction, suggesting that amino acids L53 and Y65 are crucial for promoter recognition and/or contact with core polymerase.

Function of mORF1 Protein as a Terminal Recognition Factor for the Linear Mitochondrial Plasmid pMLP1 from Pleurotus ostreatus: Eun-Kyoung Kim , Jung-Hye Roe; J. Microbiol. 1999;37(4):229-233.

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Abstract: The mitochondrial plasmid pMLP1 from a white-rot fungus, Pleurotus ostreatus, is a double-stranded DNA containing 381 bp terminal inverted repeat (TIR) whose 5'-ends are covalently bound by terminal proteins. The plasmid contains two major open reading frames (ORFs), encoding putative DNA and RNA polymerases, and a minor ORF encoding a small, highly basic protein. To identify the DNA binding activity that recognizes the TIR region of pMLP1, gel retardation assays were performed with mitochondrial extracts. A specific protein binding to a region between 123 and 248 nt within TIR was observed. We examined whether the gene product of mORF1 bindes to this region specifically. E. coli cell extract which contains an overproduced mORF1 protein formed a complex specific to the region between 123 and 248 nt. Inclusion of mORF1 protein in the specific complex formed between P. ostreatus mitochondrial extract and TIR was confirmed by a supershift assay using polyclonal antibodies against the mORF1 protein. Our result suggest that the product of mORF1 may function as a terminal region recognition factor (TRF), recognizing an internal region in TIR.

Expression of Human Mitochondiral Aldehyde Dehydrogenase 2 in Mammalian Cells using Vaccinia Virus-T7 RNA Polymerase: Kang, Su Min , Yoo, Seung Ku , Lee, Ki Whan; J. Microbiol. 1999;37(1):41-44.

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Abstract: Human mitochondrial aldehyde dehydrogenase 2 (ALDH2) is mainly responsible for oxidation of acetaldehyde generated during alcohol oxidation in vivo. A full-length cDNA of human liver ALDH2 was successfully expressed using a vaccinia virus-T7 RNA polymerase system. The expressed ALDH2 had an enzymatic activity as high as the native human liver ALDH2 enzyme.

Isolation and Characterization of the sod2+ Gene Encoding a Putative Mitochondrial Manganese Superoxide Dismutase in Schizosaccharomyces pombe: Jae-Hoon Jeong , Eun-Soo Kwon , Jung-Hye Roe; J. Microbiol. 2001;39(1):37-41.

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Abstract: The fission yeast Schizosaccharomyces pombe contains two distinct superoxide dismutase (SOD) activities, one in the cytosol encoded by the sod1+ gene and the other in mitochondria. The sod2+ gene encoding putative mitochondrial manganese superoxide dismutase (MnSOD) was isolated from the S. pombe genomic library using a PCR fragment as the probe. The nucleotide sequence of the sod2+ gene and its flanking region (4051 bp HindIII fragment) was determined. An intron of 123 nt in size was predicted and confirmed by sequencing the cDNA following reverse transcription PCR. The predicted Sod2p consists of 218 amino acid residues with a molecular mass of 24,346 Da. The deduced amino acid sequence showed a high degree of homology with other MnSODs, especially in the metal binding residues at the active site and their relative positions. The transcriptional start site was mapped by primer extension at 231 nt upstream from the ATG codon. A putative TATA box (TATAAAA) was located 58 nt upstream from the transcriptional start site and putative polyadenylation sites were located at 1000, 1062, and 1074 nt downstream from the ATG start codon.

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