Difference between revisions of "Part:BBa K3416032"
 
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===<i>lysECD7</i>===
 
===<i>lysECD7</i>===
 
=Introduction=
 
=Introduction=
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Vilnius-Lithuania iGEM 2020 project [https://2020.igem.org/Team:Vilnius-Lithuania <b>FlavoFlow]</b>includes three goals towards looking for  <i>Flavobacterium</i> disease-related problems’ solutions. The project includes creating a rapid detection kit, based on HDA and LFA, developing an implement for treating a disease, and introducing the foundation of edible vaccines.
 
Vilnius-Lithuania iGEM 2020 project [https://2020.igem.org/Team:Vilnius-Lithuania <b>FlavoFlow]</b>includes three goals towards looking for  <i>Flavobacterium</i> disease-related problems’ solutions. The project includes creating a rapid detection kit, based on HDA and LFA, developing an implement for treating a disease, and introducing the foundation of edible vaccines.
 
This part was used for the second goal- treatment -  of the project FlavoFlow.
 
This part was used for the second goal- treatment -  of the project FlavoFlow.
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==Biology==
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=Biology=
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Bacteriophage lysins are enzymes encoded by bacteriophages that cut peptidoglycans in bacterial cell walls(Fig.1). Endolysins are missing the signal peptide sequence, that is why they are not transferred through the cytoplasmic membrane1 . These enzymes have a high rate of lytic action, they also can work upon antibiotic-resistant bacterial strains as well as their ability to degrade bacterial biofilms. LysECD7 is an endolysin of the putative zinc D-alanyl-D-alanine carboxypeptidase (peptidase M15 family) of the lytic bacteriophage ECD72. This bacteriophage is related to the Myoviridae family(Fig. 2) and infects the <i>Escherichia species</i><sup>3</sup>. Normally, endolysins act from within the bacterial host at the end of the bacteriophage’s replicative cycle, allowing the release of viral offspring.  
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Bacteriophage lysins are enzymes encoded by bacteriophages that cut peptidoglycans in bacterial cell walls(Fig.1). Endolysins are missing the signal peptide sequence, that is why they are not transferred through the cytoplasmic membrane1 . These enzymes have a high rate of lytic action, they also can work upon antibiotic-resistant bacterial strains as well as their ability to degrade bacterial biofilms. LysECD7 is an endolysin of the putative zinc D-alanyl-D-alanine carboxypeptidase (peptidase M15 family) of the lytic bacteriophage ECD72. This bacteriophage is related to the Myoviridae family(Fig. 2) and infects the <i>Escherichia species</i><ref name ="Third">Myoviridae. <i>Virus Taxonomy</i> 46–62 (Elsevier, 2012).</ref>. Normally, endolysins act from within the bacterial host at the end of the bacteriophage’s replicative cycle, allowing the release of viral offspring.  
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[[File:T--Vilnius-Lithuania--endo.png|thumb|400px|centre|<b>Figure 1.</b> Endolysin activity mechanism<sup>5</sup>. a) Endolysin can not access peptidoglycan. b)
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[[File:T--Vilnius-Lithuania--endo.png|thumb|400px|centre|<b>Figure 1.</b> Endolysin activity mechanism<ref name="Fifth">Schmelcher, M. & Loessner, M. J. Bacteriophage endolysins: applications for food safety. <i>Current Opinion in Biotechnology</i> <b>37</b>, 76–87 (2016).</ref>. a) Endolysin can not access peptidoglycan. b)
 
Endolysins are able penetrate the outer membrane because: 1) they have specific membrane passing abilities; 2) they work together with permeabilizing reagents to enter through the outer membrane; 3) they are fused with self-promoted uptake mediators.  ]]
 
Endolysins are able penetrate the outer membrane because: 1) they have specific membrane passing abilities; 2) they work together with permeabilizing reagents to enter through the outer membrane; 3) they are fused with self-promoted uptake mediators.  ]]
  
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LysECD7 might become a potential therapeutic agent for antimicrobial drugs, since results revealed a broad spectrum of activity against Gram-negative bacteria<sup>4,5</sup>. However, this endolysin action depends on a lot of aspects such as buffer conditions, salts concentrations, the pH levels of the medium. For the most efficient use of endolysins, it is important that they maintain their activity under these conditions: high salt composition and pH from 5.5 to 7.42.
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LysECD7 might become a potential therapeutic agent for antimicrobial drugs, since results revealed a broad spectrum of activity against Gram-negative bacteria<ref>Antonova, N. P. et al. Modulation of Endolysin LysECD7 Bactericidal Activity by Different Peptide Tag Fusion. <i>Biomolecules</i> <b>10</b>, 440 (2020).</ref>,<ref name="Fifth"></ref>. However, this endolysin action depends on a lot of aspects such as buffer conditions, salts concentrations, the pH levels of the medium. For the most efficient use of endolysins, it is important that they maintain their activity under these conditions: high salt composition and pH from 5.5 to 7.42.
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[[File:T--Vilnius-Lithuania--virus.png|thumb|400px|centre|<b>Figure 2.</b> Typical structure of a myovirus<ref name ="Third">3</ref>. ]]
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K3416000 SequenceAndFeatures</partinfo>
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===Functional Parameters===
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<partinfo>BBa_K3416000 parameters</partinfo>
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[[File:T--Vilnius-Lithuania--virus.png|thumb|400px|centre|<b>Figure 2.</b> Typical structure of a myovirus<sup>3</sup>. ]]
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=References=
  
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===References===
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<references />
 
#Jarábková, V., Tišáková, L. & Godány, A. Phage Endolysin: A Way To Understand A Binding Function Of C-Terminal Domains, A Mini Review. <i>Nova Biotechnologica et Chimica</i> <b>14</b>, 117–134 (2015).
 
#Jarábková, V., Tišáková, L. & Godány, A. Phage Endolysin: A Way To Understand A Binding Function Of C-Terminal Domains, A Mini Review. <i>Nova Biotechnologica et Chimica</i> <b>14</b>, 117–134 (2015).
 
#Antonova, N. et al. Broad Bactericidal Activity of the Myoviridae Bacteriophage Lysins LysAm24, LysECD7, and LysSi3 against Gram-Negative ESKAPE Pathogens. <i>Viruses</i> <b>11</b>, 284 (2019).
 
#Antonova, N. et al. Broad Bactericidal Activity of the Myoviridae Bacteriophage Lysins LysAm24, LysECD7, and LysSi3 against Gram-Negative ESKAPE Pathogens. <i>Viruses</i> <b>11</b>, 284 (2019).
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#Myoviridae. <i>Virus Taxonomy</i> 46–62 (Elsevier, 2012).
 
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#Antonova, N. P. et al. Modulation of Endolysin LysECD7 Bactericidal Activity by Different Peptide Tag Fusion. <i>Biomolecules</i> <b>10</b>, 440 (2020).
 
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#Schmelcher, M. & Loessner, M. J. Bacteriophage endolysins: applications for food safety. <i>Current Opinion in Biotechnology</i> <b>37</b>, 76–87 (2016).
 

Latest revision as of 22:36, 16 December 2020

lysECD7

Introduction

FlavoFlow

Vilnius-Lithuania iGEM 2020 project FlavoFlowincludes three goals towards looking for Flavobacterium disease-related problems’ solutions. The project includes creating a rapid detection kit, based on HDA and LFA, developing an implement for treating a disease, and introducing the foundation of edible vaccines. This part was used for the second goal- treatment - of the project FlavoFlow.

Biology

Bacteriophage lysins are enzymes encoded by bacteriophages that cut peptidoglycans in bacterial cell walls(Fig.1). Endolysins are missing the signal peptide sequence, that is why they are not transferred through the cytoplasmic membrane1 . These enzymes have a high rate of lytic action, they also can work upon antibiotic-resistant bacterial strains as well as their ability to degrade bacterial biofilms. LysECD7 is an endolysin of the putative zinc D-alanyl-D-alanine carboxypeptidase (peptidase M15 family) of the lytic bacteriophage ECD72. This bacteriophage is related to the Myoviridae family(Fig. 2) and infects the Escherichia species[1]. Normally, endolysins act from within the bacterial host at the end of the bacteriophage’s replicative cycle, allowing the release of viral offspring.

Figure 1. Endolysin activity mechanism[2]. a) Endolysin can not access peptidoglycan. b) Endolysins are able penetrate the outer membrane because: 1) they have specific membrane passing abilities; 2) they work together with permeabilizing reagents to enter through the outer membrane; 3) they are fused with self-promoted uptake mediators.

LysECD7 might become a potential therapeutic agent for antimicrobial drugs, since results revealed a broad spectrum of activity against Gram-negative bacteria[3],[2]. However, this endolysin action depends on a lot of aspects such as buffer conditions, salts concentrations, the pH levels of the medium. For the most efficient use of endolysins, it is important that they maintain their activity under these conditions: high salt composition and pH from 5.5 to 7.42.


Figure 2. Typical structure of a myovirus[1].

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


References

  1. ↑ 1.0 1.1 Myoviridae. Virus Taxonomy 46–62 (Elsevier, 2012).
  2. ↑ 2.0 2.1 Schmelcher, M. & Loessner, M. J. Bacteriophage endolysins: applications for food safety. Current Opinion in Biotechnology 37, 76–87 (2016).
  3. ↑ Antonova, N. P. et al. Modulation of Endolysin LysECD7 Bactericidal Activity by Different Peptide Tag Fusion. Biomolecules 10, 440 (2020).
  1. Jarábková, V., Tišáková, L. & Godány, A. Phage Endolysin: A Way To Understand A Binding Function Of C-Terminal Domains, A Mini Review. Nova Biotechnologica et Chimica 14, 117–134 (2015).
  2. Antonova, N. et al. Broad Bactericidal Activity of the Myoviridae Bacteriophage Lysins LysAm24, LysECD7, and LysSi3 against Gram-Negative ESKAPE Pathogens. Viruses 11, 284 (2019).