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Eradication of drug-resistant Acinetobacter baumannii by cell-penetrating peptide fused endolysin
Jeonghyun Lim , Jaeyeon Jang , Heejoon Myung , Miryoung Song
J. Microbiol. 2022;60(8):859-866.   Published online May 25, 2022
DOI: https://doi.org/10.1007/s12275-022-2107-y
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AbstractAbstract
Antimicrobial agents targeting peptidoglycan have shown successful results in eliminating bacteria with high selective toxicity. Bacteriophage encoded endolysin as an alternative antibiotics is a peptidoglycan degrading enzyme with a low rate of resistance. Here, the engineered endolysin was developed to defeat multiple drug-resistant (MDR) Acinetobacter baumannii. First, putative endolysin PA90 was predicted by genome analysis of isolated Pseudomonas phage PBPA. The His-tagged PA90 was purified from BL21(DE3) pLysS and tested for the enzymatic activity using Gram-negative pathogens known for having a high antibiotic resistance rate including A. baumannii. Since the measured activity of PA90 was low, probably due to the outer membrane, cell-penetrating peptide (CPP) DS4.3 was introduced at the N-terminus of PA90 to aid access to its substrate. This engineered endolysin, DS-PA90, completely killed A. baumannii at 0.25 μM, at which concentration PA90 could only eliminate less than one log in CFU/ml. Additionally, DS-PA90 has tolerance to NaCl, where the ~50% of activity could be maintained in the presence of 150 mM NaCl, and stable activity was also observed with changes in pH or temperature. Even MDR A. baumannii strains were highly susceptible to DS-PA90 treatment: five out of nine strains were entirely killed and four strains were reduced by 3–4 log in CFU/ml. Consequently, DS-PA90 could protect waxworm from A. baumannii-induced death by ~70% for ATCC 17978 or ~44% for MDR strain 1656-2 infection. Collectively, our data suggest that CPP-fused endolysin can be an effective antibacterial agent against Gramnegative pathogens regardless of antibiotics resistance mechanisms.

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  • Antimicrobial peptide thanatin fused endolysin PA90 (Tha-PA90) for the control of Acinetobacter baumannii infection in mouse model
    Jeonghyun Lim, Heejoon Myung, Daejin Lim, Miryoung Song
    Journal of Biomedical Science.2024;[Epub]     CrossRef
  • Tissue damage alleviation and mucin inhibition by P5 in a respiratory infection mouse model with multidrug-resistant Acinetobacter baumannii
    Jun Hee Oh, Jonggwan Park, Hee Kyoung Kang, Hee Joo Park, Yoonkyung Park
    Biomedicine & Pharmacotherapy.2024; 181: 117724.     CrossRef
  • Potential of antimicrobial peptide-fused endolysin LysC02 as therapeutics for infections and disinfectants for food contact surfaces to control Cronobacter sakazakii
    Doyeon Kim, Jinwoo Kim, Minsik Kim
    Food Control.2024; 157: 110190.     CrossRef
  • Gram-negative endolysins: overcoming the outer membrane obstacle
    Hazel M Sisson, Simon A Jackson, Robert D Fagerlund, Suzanne L Warring, Peter C Fineran
    Current Opinion in Microbiology.2024; 78: 102433.     CrossRef
  • LysJEP8: A promising novel endolysin for combating multidrug‐resistant Gram‐negative bacteria
    Jose Vicente Carratalá, Neus Ferrer‐Miralles, Elena Garcia‐Fruitós, Anna Arís
    Microbial Biotechnology.2024;[Epub]     CrossRef
  • You get what you test for: The killing effect of phage lysins is highly dependent on buffer tonicity and ionic strength
    Roberto Vázquez, Diana Gutiérrez, Zoë Dezutter, Bjorn Criel, Philippe de Groote, Yves Briers
    Microbial Biotechnology.2024;[Epub]     CrossRef
  • Endolysins: a new antimicrobial agent against antimicrobial resistance. Strategies and opportunities in overcoming the challenges of endolysins against Gram-negative bacteria
    Fazal Mehmood Khan, Fazal Rasheed, Yunlan Yang, Bin Liu, Rui Zhang
    Frontiers in Pharmacology.2024;[Epub]     CrossRef
  • Characterization of Three Different Endolysins Effective against Gram-Negative Bacteria
    Tae-Hwan Jeong, Hye-Won Hong, Min Soo Kim, Miryoung Song, Heejoon Myung
    Viruses.2023; 15(3): 679.     CrossRef
  • Design strategies for positively charged endolysins: Insights into Artilysin development
    Jose Vicente Carratalá, Anna Arís, Elena Garcia-Fruitós, Neus Ferrer-Miralles
    Biotechnology Advances.2023; 69: 108250.     CrossRef
Promoter exchange of the cryptic nonribosomal peptide synthetase gene for oligopeptide production in Aspergillus oryzae
Chanikul Chutrakul , Sarocha Panchanawaporn , Sukanya Jeennor , Jutamas Anantayanon , Kobkul Laoteng
J. Microbiol. 2022;60(1):47-56.   Published online November 9, 2021
DOI: https://doi.org/10.1007/s12275-022-1442-3
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AbstractAbstract
Oligopeptides with functional activities are of current interest in the nutraceutical and medical sectors. The development of the biosynthetic process of oligopeptides through a nonribosomal peptide synthetase (NRPS) system has become more challenging. To develop a production platform for nonribosomal peptides (NRPs), reprogramming of transcriptional regulation of the acv gene encoded ACV synthetase (ACVS) was implemented in Aspergillus oryzae using the CRISPRCas9 system. Awakening silent acv expression was successfully achieved by promoter substitution. Among the three exchanged promoters, AoPgpdA, AoPtef1, and PtPtoxA, the replacement of the native promoter with AoPgpdA led to the highest ACV production in A. oryzae. However, the ACV production of the AoPGpdA strain was also dependent on the medium composition, in which urea was the best nitrogen source, and a C:N ratio of 20:1 was optimal for tripeptide production. In addition to cell growth, magnesium ions are an essential element for ACV production and might participate in ACVS activity. It was also found that ACV was the growthassociated product of the engineered strain that might be a
result
of constitutive transcriptional control by the AoPgpdA promoter. This study offers a potential strategy for nonribosomal ACV production using the fungal system, which is applicable for redesigning bioactive oligopeptides with industrial relevance.

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  • Strategies for the Enhancement of Secondary Metabolite Production via Biosynthesis Gene Cluster Regulation in Aspergillus oryzae
    Xiao Jia, Jiayi Song, Yijian Wu, Sai Feng, Zeao Sun, Yan Hu, Mengxue Yu, Rui Han, Bin Zeng
    Journal of Fungi.2024; 10(5): 312.     CrossRef
  • Transcriptome-based Mining of the Constitutive Promoters for Tuning Gene Expression in Aspergillus oryzae
    Kobkul Laoteng, Jutamas Anantayanon, Chanikul Chutrakul, Sarocha Panchanawaporn, Sukanya Jeennor
    Journal of Microbiology.2023; 61(2): 199.     CrossRef
  • Efficient de novo production of bioactive cordycepin by Aspergillus oryzae using a food-grade expression platform
    Sukanya Jeennor, Jutamas Anantayanon, Sarocha Panchanawaporn, Chanikul Chutrakul, Wanwipa Vongsangnak, Kobkul Laoteng
    Microbial Cell Factories.2023;[Epub]     CrossRef
  • Synthetic microbes and biocatalyst designs in Thailand
    Duangthip Trisrivirat, Ruchanok Tinikul, Pimchai Chaiyen
    Biotechnology Notes.2023; 4: 28.     CrossRef
  • Potential of Aspergillus oryzae as a biosynthetic platform for indigoidine, a non-ribosomal peptide pigment with antioxidant activity
    Sarocha Panchanawaporn, Chanikul Chutrakul, Sukanya Jeennor, Jutamas Anantayanon, Nakul Rattanaphan, Kobkul Laoteng, Daniel Cullen
    PLOS ONE.2022; 17(6): e0270359.     CrossRef
  • CRISPR/Cas9-Based Genome Editing and Its Application in Aspergillus Species
    Feng-Jie Jin, Bao-Teng Wang, Zhen-Dong Wang, Long Jin, Pei Han
    Journal of Fungi.2022; 8(5): 467.     CrossRef
Research Support, Non-U.S. Gov'ts
Antimicrobial Resistance, Virulence Genes and PFGE-profiling of Escherichia coli Isolates from South Korean Cattle Farms
Seung Won Shin , Jae-Won Byun , Myounghwan Jung , Min-Kyoung Shin , Han Sang Yoo
J. Microbiol. 2014;52(9):785-793.   Published online July 30, 2014
DOI: https://doi.org/10.1007/s12275-014-4166-1
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AbstractAbstract
To estimate the prevalence of Escherichia coli with potential pathogenicity in cattle farm in South Korea, a total of 290 E. coli isolates were isolated from cattle farms over a period of 2 years in South Korea. These were examined for phenotypic and genotypic characteristics including antimicrobial susceptibility, serotype, and gene profiles of virulence and antimicrobial resistance. The most dominant virulence gene was f17 (26.2%), followed by stx2 (15.9%), ehxA (11.0%), stx1 (8.3%), eae (5.2%), and sta (4.1%). Some shiga-toxin producing E. coli isolates possessed eae (15.9%). All isolates except for one showed resistance to one or more antimicrobials, with 152 isolates exhibiting multidrug-resistance. The most prevalent resistance phenotype detected was streptomycin (63.1%), followed by tetracycline (54.5%), neomycin (40.3%), cephalothin (32.8%), amoxicillin (30.0%), ampicillin (29.7%), and sulphamethoxazole/trimethoprim (16.6%). The associated resistance determinants detected were strAstrB (39.0%), tet(E) (80.0%), tet(A) (27.6%), aac(3)-IV (33.1%), aphA1 (21.4%), blaTEM (23.8%), and sul2 (22.1%). When investigated by O serotyping and PFGE molecular subtyping, the high degree of diversity was exhibited in E. coli isolates. These results suggest that E. coli isolates from South Korean cattle farms are significantly diverse in terms of virulence and antimicrobial resistance. In conclusion, the gastroinstestinal flora of cattle could be a significant reservoir of diverse virulence and antimicrobial resistance determinants, which is potentially hazardous to public health.

Citations

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  • Underrepresented high diversity of class 1 integrons in the environment uncovered by PacBio sequencing using a new primer
    Yu Yang, An-Ni Zhang, You Che, Lei Liu, Yu Deng, Tong Zhang
    Science of The Total Environment.2021; 787: 147611.     CrossRef
  • The prevalence of causative agents of calf diarrhea in Korean native calves
    Jeong-Byoung Chae, Hyeon-Cheol Kim, Jun-Gu Kang, Kyoung-Seong Choi, Joon-Seok Chae, Do-Hyeon Yu, Bae-Keun Park, Yeon-su Oh, Hak-Jong Choi, Jinho Park
    Journal of Animal Science and Technology.2021; 63(4): 864.     CrossRef
  • O-serogroups, virulence genes, antimicrobial susceptibility, and MLST genotypes of Shiga toxin-producing Escherichia coli from swine and cattle in Central China
    Zhong Peng, Wan Liang, Zizhe Hu, Xiaosong Li, Rui Guo, Lin Hua, Xibiao Tang, Chen Tan, Huanchun Chen, Xiangru Wang, Bin Wu
    BMC Veterinary Research.2019;[Epub]     CrossRef
  • Detection and molecular characterization of sorbitol fermenting non-O157 Escherichia coli from goats
    Shivasharanappa Nayakvadi, Charlotte Alison Alemao, H.B. Chethan Kumar, R.S. Rajkumar, Susitha Rajkumar, Eaknath B. Chakurkar, Shivaramu Keelara
    Small Ruminant Research.2018; 161: 7.     CrossRef
  • Antimicrobial Resistance inEscherichia coli
    Laurent Poirel, Jean-Yves Madec, Agnese Lupo, Anne-Kathrin Schink, Nicolas Kieffer, Patrice Nordmann, Stefan Schwarz, Frank Møller Aarestrup, Stefan Schwarz, Jianzhong Shen, Lina Cavaco
    Microbiology Spectrum.2018;[Epub]     CrossRef
  • The genetic background of antibiotic resistance among clinical uropathogenic Escherichia coli strains
    Wioletta Adamus-Białek, Anna Baraniak, Monika Wawszczak, Stanisław Głuszek, Beata Gad, Klaudia Wróbel, Paulina Bator, Marta Majchrzak, Paweł Parniewski
    Molecular Biology Reports.2018; 45(5): 1055.     CrossRef
  • Interrelationship between tetracycline resistance determinants, phylogenetic group affiliation and carriage of class 1 integrons in commensal Escherichia coli isolates from cattle farms
    Kuastros Mekonnen Belaynehe, Seung Won Shin, Han Sang Yoo
    BMC Veterinary Research.2018;[Epub]     CrossRef
  • Occurrence of aminoglycoside-modifying enzymes among isolates of Escherichia coli exhibiting high levels of aminoglycoside resistance isolated from Korean cattle farms
    Kuastros Mekonnen Belaynehe, Seung Won Shin, Park Hong-Tae, Han Sang Yoo
    FEMS Microbiology Letters.2017;[Epub]     CrossRef
  • Prevalence of Antimicrobial Resistance and Transfer of Tetracycline Resistance Genes in Escherichia coli Isolates from Beef Cattle
    Seung Won Shin, Min Kyoung Shin, Myunghwan Jung, Kuastros Mekonnen Belaynehe, Han Sang Yoo, M. W. Griffiths
    Applied and Environmental Microbiology.2015; 81(16): 5560.     CrossRef
  • Profiling of antimicrobial resistance and plasmid replicon types in β-lactamase producingEscherichia coliisolated from Korean beef cattle
    Seung Won Shin, Myunghwan Jung, Min-Kyung Shin, Han Sang Yoo
    Journal of Veterinary Science.2015; 16(4): 483.     CrossRef
Prevalence of Human Papillomavirus Infection and Genotype Distribution Determined by the Cyclic-Catcher Melting Temperature Analysis in Korean Medical Checkup Population
Yun-Jee Kim , Min-Jung Kwon , Hee-Yeon Woo , Soon-Young Paik
J. Microbiol. 2013;51(5):665-670.   Published online September 14, 2013
DOI: https://doi.org/10.1007/s12275-013-3160-3
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AbstractAbstract
Although cytology screening has reduced the incidence and mortality rate of cervical cancer significantly, its usefulness is limited to samples from the site of the lesion, resulting in its low sensitivity and unsuitability for use in medical checkups. The purpose of the present study was to evaluate the prevalence of HPV infection using genotype distribution and to analyze the correlation of the HPV DNA test results with cytological results. We also evaluated the benefits of quantitative information obtained from cyclic-catcher melting temperature analysis (CMTA) in screening for cervical cancer. We performed cyclic-CMTA using AnyplexTM II HPV28 Detection in combination with cervical cytology for 2,181 subjects. The following HPV positivity types were detected using cyclic-CMTA and HPV positivity was found to increase together with the severity of the cytology results: (1) For 419 HPV positive specimens, HPV DNA was detected in 18.1% of normal specimens, 78.3% of ASCUS, and all of LSIL and HSIL; (2) high-risk HPV DNAs were detected in 63.3% of normal (N=547), 65.9% of ASCUS (N=41), 76.9% of LSIL (N=13), and 88.9% of HSIL (N=9) among total detected HPV DNA regardless multiple detection; (3) multiple HPV genotypes were detected in 4.8% of normal specimens (N=2,146), 52.2% of ASCUS (N=23), 57.1% of LSIL (N=7), and 40.0% of HSIL (N=5). In addition, a high level of viral DNA was observed using cyclic-CMTA in all specimens beyond the LSIL stage according to cytology, while only 6% of specimens with normal cytology showed a correlation with viral quantitation by cyclic-CMTA. The combination of HPV genotyping with a quantitative assay and cytology will allow for a more accurate diagnosis of cervical cancer.

Citations

Citations to this article as recorded by  
  • Comparative evaluation of HPV genotyping: A study on the performance concordance between Anyplex II HPV28 detection and Linear Array genotyping tests in nationwide studies in Brazil
    Isabel Cristina Bandeira, Juliana Comerlato, Marina Bessel, Bruna Vieira Fernandes, Giana Mota, Luisa Lina Villa, Flávia Moreno Alves de Souza, Gerson Fernando Mendes Pereira, Eliana Marcia Wendland, Hideo Kato
    PLOS ONE.2024; 19(6): e0305122.     CrossRef
  • Use of swabs for dry collection of self-samples to detect human papillomavirus among Malagasy women
    Pierre Vassilakos, Rosa Catarino, Stephanie Bougel, Maria Munoz, Caroline Benski, Ulrike Meyer-Hamme, Jeromine Jinoro, Josea Lea Heriniainasolo, Patrick Petignat
    Infectious Agents and Cancer.2016;[Epub]     CrossRef
  • Clinical Validation of Anyplex II HPV HR Detection Test for Cervical Cancer Screening in Korea
    Sunkyung Jung, Byungdoo Lee, Kap No Lee, Yonggoo Kim, Eun-Jee Oh
    Archives of Pathology & Laboratory Medicine.2016; 140(3): 276.     CrossRef
  • Therapeutic potential of an AcHERV-HPV L1 DNA vaccine
    Hee-Jung Lee, Jong Kwang Yoon, Yoonki Heo, Hansam Cho, Yeondong Cho, Yongdae Gwon, Kang Chang Kim, Jiwon Choi, Jae Sung Lee, Yu-Kyoung Oh, Young Bong Kim
    Journal of Microbiology.2015; 53(6): 415.     CrossRef
Distribution of Marine Birnavirus in Cultured Olive Flounder Paralichthys olivaceus in Korea
Sung-Ju Jung , Seok-Ryel Kim , Il-Yong Joung , Shin-Ichi Kitamura , Hee-Taek Ceong , Myung-Joo Oh
J. Microbiol. 2008;46(3):265-273.   Published online July 5, 2008
DOI: https://doi.org/10.1007/s12275-008-0004-7
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AbstractAbstract
Surveys of marine birnavirus (MABV) were undertaken in cultured olive flounder Paralichthys olivaceus from the south and west coastal areas and Jeju in Korea during the period January 1999 to April 2007. MABV was detected in all seasons from the fry, juveniles and adult fish from the areas examined. Evident cytopathic effects of the virus including rounding and cell lysis were observed in chinook salmon embryo (CHSE-214) and rainbow trout gonad (RTG-2) cells, but not in fathead minnow (FHM) and epithelial papilloma of carp (EPC) cells. Nucleotide sequences of the VP2/NS junction region of the Korean isolates showed 97.8%~100% similarity, and they belonged to the same genogroup. Cross neutralization tests with serotype-specific rabbit antisera against MABV strains exhibited a close antigenic relationships between strains, and were distinct from infectious pancreatic necrosis virus (IPNV) strains. Coinfection of MABV with bacteria (Streptococcus iniae, Vibrio spp.) and viruses (nervous necrosis virus, lymphocystis disease virus, viral hemorrhagic septicemia virus) was observed.
Prevalence and Antibiotic Susceptibility of Vancomycin-Resistant Enterococci in Chicken Intestines and Fecal Samples from Healthy Young Children and Intensive Care Unit Patients
Shin Moo Kim , Eun Sook Shim , Chi Nam Seong
J. Microbiol. 2001;39(2):116-120.
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AbstractAbstract
The prevalence, resistance genotype and antibiotic susceptibility of vancomycin-resistant enterococci (VRE) were determined. Prevalence of VRE in chickens, healthy children and intensive care unit (ICU) patients was 43.0%, 12.7% and 24.1%, respectively. Forty out of 56 isolates from chicken intestines were identified as Enterococcus faecium, and 12 were E. faecalis. All the isolates contained the vanA gene. Nine out of 13 VRE isolates from patients and two out of 21 from healthy young children were identified as E. faecium. The resistance types of E. faecium, E. gallinarium and E. casseliflavus were VanA, VanC1, and VanC2, respectively. The mimimum inhibitory concentrations (MICs) of E. faecium, E. gallinarium, and E. casseliflavus to vancomycin were 512, 8 and 4 g/ml, respectively. Specifically, E. faecium isolates were resistant to most of antibiotics except ampicillin and gentamicin. This is the first report of high VanA type VRE prevalence in nonhospitalized young children in Korea.

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