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2 "Constitutive promoter"
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Characterization of a novel phage depolymerase specific to Escherichia coli O157:H7 and biofilm control on abiotic surfaces
Do-Won Park , Jong-Hyun Park
J. Microbiol. 2021;59(11):1002-1009.   Published online October 6, 2021
DOI: https://doi.org/10.1007/s12275-021-1413-0
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AbstractAbstract
The increasing prevalence of foodborne diseases caused by Escherichia coli O157:H7 as well as its ability to form biofilms poses major threats to public health worldwide. With increasing concerns about the limitations of current disinfectant treatments, phage-derived depolymerases may be used as promising biocontrol agents. Therefore, in this study, the characterization, purification, and application of a novel phage depolymerase, Dpo10, specifically targeting the lipopolysaccharides of E. coli O157, was performed. Dpo10, with a molecular mass of 98 kDa, was predicted to possess pectate lyase activity via genome analysis and considered to act as a receptor- binding protein of the phage. We confirmed that the purified Dpo10 showed O-polysaccharide degrading activity only for the E. coli O157 strains by observing its opaque halo. Dpo10 maintained stable enzymatic activities across a wide range of temperature conditions under 55°C and mild basic pH. Notably, Dpo10 did not inhibit bacterial growth but significantly increased the complement-mediated serum lysis of E. coli O157 by degrading its O-polysaccharides. Moreover, Dpo10 inhibited the biofilm formation against E. coli O157 on abiotic polystyrene by 8-fold and stainless steel by 2.56 log CFU/coupon. This inhibition was visually confirmed via fieldemission scanning electron microscopy. Therefore, the novel depolymerase from E. coli siphophage exhibits specific binding and lytic activities on the lipopolysaccharide of E. coli O157 and may be used as a promising anti-biofilm agent against the E. coli O157:H7 strain.

Citations

Citations to this article as recorded by  
  • Effect of Bacteriophages against Biofilms of Escherichia coli on Food Processing Surfaces
    Ana Brás, Márcia Braz, Inês Martinho, João Duarte, Carla Pereira, Adelaide Almeida
    Microorganisms.2024; 12(2): 366.     CrossRef
  • Bacteriophage–Host Interactions and the Therapeutic Potential of Bacteriophages
    Leon M. T. Dicks, Wian Vermeulen
    Viruses.2024; 16(3): 478.     CrossRef
  • Current Strategies for Combating Biofilm-Forming Pathogens in Clinical Healthcare-Associated Infections
    Rashmita Biswas, Bhawana Jangra, Ganapathy Ashok, Velayutham Ravichandiran, Utpal Mohan
    Indian Journal of Microbiology.2024; 64(3): 781.     CrossRef
  • Phage Adsorption to Gram-Positive Bacteria
    Audrey Leprince, Jacques Mahillon
    Viruses.2023; 15(1): 196.     CrossRef
  • Prevalence of Indigenous Antibiotic-Resistant Salmonella Isolates and Their Application to Explore a Lytic Phage vB_SalS_KFSSM with an Intra-Broad Specificity
    Jaein Choe, Su-Hyeon Kim, Ji Min Han, Jong-Hoon Kim, Mi-Sun Kwak, Do-Won Jeong, Mi-Kyung Park
    Journal of Microbiology.2023; 61(12): 1063.     CrossRef
  • Phages against Pathogenic Bacterial Biofilms and Biofilm-Based Infections: A Review
    Siyu Liu, Hongyun Lu, Shengliang Zhang, Ying Shi, Qihe Chen
    Pharmaceutics.2022; 14(2): 427.     CrossRef
Evaluation and application of constitutive promoters for cutinase production by Saccharomyces cerevisiae
Juan Zhang , Yanqiu Cai , Guocheng Du , Jian Chen , Miao Wang , Zhen Kang
J. Microbiol. 2017;55(7):538-544.   Published online June 30, 2017
DOI: https://doi.org/10.1007/s12275-017-6514-4
  • 47 View
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  • 5 Crossref
AbstractAbstract
died and applied in processes targeted for industrial scale. In this work, the cutinase gene tfu from Thermobifida fusca was artificially synthesized according to codon usage bias of Saccharomyces cerevisiae and investigated in Saccharomyces cerevisiae. Using the α-factor signal peptide, the T. fusca cutinase was successfully overexpressed and secreted with the GAL1 expression system. To increase the cutinase level and overcome some of the drawbacks of induction, four different strong promoters (ADH1, HXT1, TEF1, and TDH3) were comparatively evaluated for cutinase production. By comparison, promoter TEF1 exhibited an outstanding property and significantly increased the expression level. By fed-batch fermentation with a constant feeding approach, the activity of cutinase was increased to 29.7 U/ml. The result will contribute to apply constitutive promoter TEF1 as a tool for targeted cutinase production in S. cerevisiae cell factory.

Citations

Citations to this article as recorded by  
  • Engineering strategies for enhanced heterologous protein production by Saccharomyces cerevisiae
    Meirong Zhao, Jianfan Ma, Lei Zhang, Haishan Qi
    Microbial Cell Factories.2024;[Epub]     CrossRef
  • Engineering the xylose metabolism of Saccharomyces cerevisiae for ethanol and single cell protein bioconversion
    Mengtian Huang, Zhuocheng Jin, Hong Ni, Peining Zhang, Huanan Li, Jiashu Liu, Chengcheng Weng, Zhengbing Jiang
    Biomass and Bioenergy.2024; 190: 107372.     CrossRef
  • An outlook to sophisticated technologies and novel developments for metabolic regulation in the Saccharomyces cerevisiae expression system
    Yijian Wu, Sai Feng, Zeao Sun, Yan Hu, Xiao Jia, Bin Zeng
    Frontiers in Bioengineering and Biotechnology.2023;[Epub]     CrossRef
  • A CRISPR–Cas9 System-Mediated Genetic Disruption and Multi-fragment Assembly in Starmerella bombicola
    Yibo Shi, Lihua Zhang, Min Zhang, Jieyu Chu, Yuanyuan Xia, Haiquan Yang, Liming Liu, Xianzhong Chen
    ACS Synthetic Biology.2022; 11(4): 1497.     CrossRef
  • Recent advances in genetic engineering tools based on synthetic biology
    Jun Ren, Jingyu Lee, Dokyun Na
    Journal of Microbiology.2020; 58(1): 1.     CrossRef

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