Saturation Mutagenesis of Lysine 12 Leads to the Identification of Derivatives of Nisin A with Enhanced Antimicrobial Activity
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{"title"=>"Saturation Mutagenesis of Lysine 12 Leads to the Identification of Derivatives of Nisin A with Enhanced Antimicrobial Activity", "type"=>"journal", "authors"=>[{"first_name"=>"Evelyn M.", "last_name"=>"Molloy", "scopus_author_id"=>"53983228800"}, {"first_name"=>"Des", "last_name"=>"Field", "scopus_author_id"=>"21233304600"}, {"first_name"=>"Paula M.", "last_name"=>"O'Connor", "scopus_author_id"=>"7402622563"}, {"first_name"=>"Paul D.", "last_name"=>"Cotter", "scopus_author_id"=>"7006716039"}, {"first_name"=>"Colin", "last_name"=>"Hill", "scopus_author_id"=>"7403558967"}, {"first_name"=>"R. Paul", "last_name"=>"Ross", "scopus_author_id"=>"55664948500"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"pmid"=>"23505531", "doi"=>"10.1371/journal.pone.0058530", "sgr"=>"84874929048", "isbn"=>"1932-6203", "scopus"=>"2-s2.0-84874929048", "issn"=>"19326203", "pui"=>"368506757"}, "id"=>"8eb0e842-c6de-3d6b-bfc0-4143a15d66f0", "abstract"=>"It is becoming increasingly apparent that innovations from the \"golden age\" of antibiotics are becoming ineffective, resulting in a pressing need for novel therapeutics. The bacteriocin family of antimicrobial peptides has attracted much attention in recent years as a source of potential alternatives. The most intensively studied bacteriocin is nisin, a broad spectrum lantibiotic that inhibits gram-positive bacteria including important food pathogens and clinically relevant antibiotic resistant bacteria. Nisin is gene-encoded and, as such, is amenable to peptide bioengineering, facilitating the generation of novel derivatives that can be screened for desirable properties. It was to this end that we used a site-saturation mutagenesis approach to create a bank of producers of nisin A derivatives that differ with respect to the identity of residue 12 (normally lysine; K12). A number of these producers exhibited enhanced bioactivity and the nisin A K12A producer was deemed of greatest interest. Subsequent investigations with the purified antimicrobial highlighted the enhanced specific activity of this modified nisin against representative target strains from the genera Streptococcus, Bacillus, Lactococcus, Enterococcus and Staphylococcus.", "link"=>"http://www.mendeley.com/research/saturation-mutagenesis-lysine-12-leads-identification-derivatives-nisin-enhanced-antimicrobial-activ", "reader_count"=>27, "reader_count_by_academic_status"=>{"Unspecified"=>2, "Researcher"=>5, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>8, "Student > Postgraduate"=>2, "Student > Master"=>5, "Student > Bachelor"=>2, "Lecturer"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>2, "Researcher"=>5, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>8, "Student > Postgraduate"=>2, "Student > Master"=>5, "Student > Bachelor"=>2, "Lecturer"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>3, "Biochemistry, Genetics and Molecular Biology"=>4, "Agricultural and Biological Sciences"=>14, "Chemical Engineering"=>1, "Chemistry"=>3, "Psychology"=>1, "Immunology and Microbiology"=>1}, "reader_count_by_subdiscipline"=>{"Chemistry"=>{"Chemistry"=>3}, "Psychology"=>{"Psychology"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>14}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>4}, "Unspecified"=>{"Unspecified"=>3}, "Chemical Engineering"=>{"Chemical Engineering"=>1}}, "reader_count_by_country"=>{"Ireland"=>1, "South Africa"=>1, "Chile"=>1, "Portugal"=>1, "India"=>1}, "group_count"=>1}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/983640"], "description"=>"<p>The structure of nisin A is depicted, showing the location of its five (β-methyl)lanthionine rings (A–E) and modified residues dehydroalanine (yellow) and dehydrobutyrine (green). Position K12 is highlighted in red and the hinge region is boxed.</p>", "links"=>[], "tags"=>["nisin"], "article_id"=>649489, "categories"=>["Biotechnology", "Genetics", "Microbiology"], "users"=>["Evelyn M. Molloy", "Des Field", "Paula M. O'. Connor", "Paul D. Cotter", "Colin Hill", "R. Paul Ross"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0058530.g001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Structure_of_nisin_A_/649489", "title"=>"Structure of nisin A.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-03-12 08:02:32"}
  • {"files"=>["https://ndownloader.figshare.com/files/983642"], "description"=>"<p>Representative RP-HPLC peaks showing the appearance of A K12S and B K12T after optimization of solvent gradient. Relative production levels were determined using triplicate peak areas from two independent purifications from first principles. Relative Standard Deviation<12% for each given value.</p>", "links"=>[], "tags"=>["nisin"], "article_id"=>649491, "categories"=>["Biotechnology", "Genetics", "Microbiology"], "users"=>["Evelyn M. Molloy", "Des Field", "Paula M. O'. Connor", "Paul D. Cotter", "Colin Hill", "R. Paul Ross"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0058530.g002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_RP_HPLC_Separation_of_nisin_A_K12S_Dha_and_K12T_Dhb_/649491", "title"=>"RP-HPLC Separation of nisin A K12S/Dha and K12T/Dhb.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-03-12 08:02:43"}
  • {"files"=>["https://ndownloader.figshare.com/files/1012135"], "description"=>"<p>Observed molecular mass (+/−0.25 Da) from MALDI-TOF MS analysis of NZ9800 pDF05-K12X producers. ND: not detected. * represents dehydrated forms (hydrophobic modified residues), i.e. Dhb in the case of T and Dha in the case of S.</p><p>Bioactivity of NZ9800 pDF05-K12X producers against <i>L. lactis</i> HP. Values given are the mean of triplicate deferred antagonism assays and represent zone of inhibition (diameter of zone minus diameter of bacterial growth) expressed as a percentage compared to that of the wild-type nisin producer at 100%. S.D.: Standard Deviation; Relative Standard Deviation <10% for each given value. N/A: not applicable. All values in bold reached statistical significance compared to nisin control (K) (Student's t-test: P<0.05).</p>", "links"=>[], "tags"=>["spectrometry", "bioactivity", "nisin", "k12x"], "article_id"=>672547, "categories"=>["Biotechnology", "Genetics", "Microbiology"], "users"=>["Evelyn M. Molloy", "Des Field", "Paula M. O'. Connor", "Paul D. Cotter", "Colin Hill", "R. Paul Ross"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0058530.t003"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Mass_Spectrometry_Analysis_and_Bioactivity_Determination_of_Nisin_K12X_Bank_/672547", "title"=>"Mass Spectrometry Analysis and Bioactivity Determination of Nisin K12X Bank.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-03-11 00:42:27"}
  • {"files"=>["https://ndownloader.figshare.com/files/1012156"], "description"=>"<p>PHO indicates 5′ phosphate modification. Underlined sequences represent degenerate codon (N = A+C+G+T, K = G+T, M = A+C). Lower-case letters indicate site-directed mutation.</p>", "links"=>[], "tags"=>["genetics and genomics", "microbiology", "biotechnology"], "article_id"=>672565, "categories"=>["Biotechnology", "Genetics", "Microbiology"], "users"=>["Evelyn M. Molloy", "Des Field", "Paula M. O'. Connor", "Paul D. Cotter", "Colin Hill", "R. Paul Ross"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0058530.t002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Oligonucleotides_used_in_this_study_/672565", "title"=>"Oligonucleotides used in this study.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-03-11 00:42:45"}
  • {"files"=>["https://ndownloader.figshare.com/files/1012176"], "description"=>"<p>DPC: Dairy Products Research Centre, Moorepark; UCC: University College Cork; NCDO: National Collection of Dairy Organisms; DSMZ: German Collection of Microorganisms and Cell Cultures; ATCC: American Type Culture Collection; NCIMB: National Collection of Industrial, Food and Marine Bacteria; BSAC: British Society for Antimicrobial Chemotherapy; ILSI: International Life Science Institute (MartinWiedmann), ARS: Agricultural Research Service, U.S. Department of Agriculture (Todd Ward).</p>", "links"=>[], "tags"=>["plasmids"], "article_id"=>672588, "categories"=>["Biotechnology", "Genetics", "Microbiology"], "users"=>["Evelyn M. Molloy", "Des Field", "Paula M. O'. Connor", "Paul D. Cotter", "Colin Hill", "R. Paul Ross"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0058530.t001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Strains_and_plasmids_used_in_this_study_/672588", "title"=>"Strains and plasmids used in this study.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-03-11 00:43:08"}
  • {"files"=>["https://ndownloader.figshare.com/files/1012194"], "description"=>"<p>Results from agarose gel diffusion assays of purified nisin and nisin K12A at a concentration of 40 µM against three Gram-negative strains. Results are expressed as both zone diameter and as K12A bioactivity compared to that of nisin A at 100%. Values represent the mean of triplicate agarose assay results. Standard deviation values in brackets; Relative Standard Deviation<10% for each given value. Values in bold reached statistical significance compared to nisin control (Student's t-test: P<3E<sup>−04</sup>).</p>", "links"=>[], "tags"=>["purified", "nisin", "k12a", "gram-negative"], "article_id"=>672604, "categories"=>["Biotechnology", "Genetics", "Microbiology"], "users"=>["Evelyn M. Molloy", "Des Field", "Paula M. O'. Connor", "Paul D. Cotter", "Colin Hill", "R. Paul Ross"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0058530.t006"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Bioactivity_of_purified_nisin_WT_and_nisin_K12A_against_representative_Gram_negative_targets_/672604", "title"=>"Bioactivity of purified nisin (WT) and nisin K12A against representative Gram-negative targets.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-03-11 00:43:24"}
  • {"files"=>["https://ndownloader.figshare.com/files/1012215"], "description"=>"<p>Results from minimum inhibitory concentration assays of purified nisin (WT) and nisin K12A against various Gram-positive targets. Values given are identical results from three independent determinations. Fold Difference represents the improvement of K12A compared to nisin against the relevant indicator.</p>", "links"=>[], "tags"=>["inhibitory", "concentrations", "purified", "nisin", "k12a", "gram-positive"], "article_id"=>672625, "categories"=>["Biotechnology", "Genetics", "Microbiology"], "users"=>["Evelyn M. Molloy", "Des Field", "Paula M. O'. Connor", "Paul D. Cotter", "Colin Hill", "R. Paul Ross"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0058530.t005"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Minimum_inhibitory_concentrations_of_purified_nisin_WT_and_nisin_K12A_against_various_Gram_positive_targets_/672625", "title"=>"Minimum inhibitory concentrations of purified nisin (WT) and nisin K12A against various Gram-positive targets.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-03-11 00:43:45"}
  • {"files"=>["https://ndownloader.figshare.com/files/1012234"], "description"=>"<p>Values are the mean of triplicate deferred antagonism assays and represent zone of inhibition (diameter of zone minus diameter of bacterial growth) expressed as a percentage compared to that of the wild-type nisin producer at 100%. S.D.: Standard Deviation; Relative Standard Deviation<10% for each given value. All values in bold reached statistical significance compared to nisin control (K) (Student's t-test: P<0.05). Strains marked with an asterisk are drug resistant isolates.</p>", "links"=>[], "tags"=>["k12s", "k12t", "producers", "gram-positive"], "article_id"=>672643, "categories"=>["Biotechnology", "Genetics", "Microbiology"], "users"=>["Evelyn M. Molloy", "Des Field", "Paula M. O'. Connor", "Paul D. Cotter", "Colin Hill", "R. Paul Ross"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0058530.t004"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Bioactivity_of_K12A_K12S_and_K12T_producers_against_various_Gram_positive_targets_/672643", "title"=>"Bioactivity of K12A, K12S and K12T producers against various Gram-positive targets.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-03-11 00:44:03"}

PMC Usage Stats | Further Information

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  • {"unique-ip"=>"2", "full-text"=>"1", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"3"}
  • {"unique-ip"=>"2", "full-text"=>"2", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"4"}
  • {"unique-ip"=>"9", "full-text"=>"9", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"5"}
  • {"unique-ip"=>"9", "full-text"=>"10", "pdf"=>"4", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"8"}
  • {"unique-ip"=>"5", "full-text"=>"4", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"9"}

Relative Metric

{"start_date"=>"2013-01-01T00:00:00Z", "end_date"=>"2013-12-31T00:00:00Z", "subject_areas"=>[{"subject_area"=>"/Biology and life sciences/Microbiology", "average_usage"=>[293, 503, 638, 755, 861, 960, 1056, 1146, 1239, 1323, 1403, 1491, 1568]}, {"subject_area"=>"/Medicine and health sciences", "average_usage"=>[264, 460, 584, 692, 794, 887, 978, 1067, 1154, 1241, 1328, 1408, 1474]}, {"subject_area"=>"/Medicine and health sciences/Pharmacology", "average_usage"=>[265, 471, 604, 723, 827, 926, 1022, 1113, 1210, 1290, 1377, 1454, 1532]}]}
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