Difference between revisions of "Part:BBa K302010"

 
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<partinfo>BBa_K302010 short</partinfo>
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<partinfo>BBa_K1152010 short</partinfo>
  
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''Bacillus subtilis'' 168 is unable to produce surfactin, a natural surfactant which helps in reducing surface tension and aids bacteria to swarm on a solid or semi solid medium. The ''sfp'' gene restores the capability to produce surfactin.
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svp encodes for a 4'-Phosphopanthetheinyl-transferase (PPTase) which transfers a 4'-PPT residue from CoA to a conserved serine residue in the T-Domain of NRPS modules, thus activating these enzymes.
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[[File:Heidelberg2013_PPtase_mechanism.png|280px|thumb|'''Figure 1:''' General mechanism of activation the activation of NRP Synthetases from apo- to holo-form.]]
  
 
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===Usage and Biology===
 
===Usage and Biology===
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Svp is a PPTase often associated with activating the indigoidine synthetase bpsA [Takahashi, 2007].
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K302010 SequenceAndFeatures</partinfo>
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<partinfo>BBa_K1152010 SequenceAndFeatures</partinfo>
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===References===
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* Takahashi, H., Kumagai, T., Kitani, K., Mori, M., Matoba, Y., & Sugiyama, M. (2007). Cloning and Characterization of a Streptomyces Single Module Type Non-ribosomal Peptide Synthetase Catalyzing a Blue Pigment Synthesis *, 282(12), 9073–9081. doi:10.1074/jbc.M611319200
  
  
 
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===Functional Parameters===
 
===Functional Parameters===
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<partinfo>BBa_K302010 parameters</partinfo>
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<partinfo>BBa_K1152010 parameters</partinfo>
 
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===Information contributed by City of London UK (2021)===
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Part information is collated here to help future users of the BioBrick registry.
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 +
Metadata:
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*'''Group:''' City of London UK 2021
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*'''Author:''' Janusan Jeyananthan
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*'''Summary:''' Added information collated from existing scientific studies
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----
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Kinetics:
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kcat is 86 min(-1) with S.glaucescens apo-ACP TcmM as the substrate and 11 min(-1) with S.mobaraensis apo-PCP BlmI as the substrate.
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For Streptomyces mobaraensis apo-PCP BlmI1: KM=3.9 µM
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For Streptomyces glaucescens apo-ACP TcmM: KM=3.1 µM
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<ref>“Svp - 4’-Phosphopantetheinyl Transferase Svp - Streptomyces Mobaraensis - Svp Gene & Protein” n.d. Uniprot. Accessed 12 August, 2021 https://www.uniprot.org/uniprot/Q9F0Q6 </ref>
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[[File:BBa_K302010.png|x200px|center]]
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===References===
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 +
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===Information contributed by NFLS_Nanjing (2022)===
 +
Part information is collated here to help future users of the BioBrick registry.
 +
 +
Metadata:
 +
*'''Group:''' NFLS_Nanjing (2022)
 +
*'''Author:''' Yuming Xue
 +
*'''Summary:''' Added information collated from existing scientific studies
 +
 +
Biology and usage:
 +
 +
1. The gene bpsA is expressed in C. glutamicum and carries strong fluxes toward L-glutamate, a precursor of indigoidine. [1]
 +
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2. The gene bpsA catalyzes indigoidine formation from two molecules of glutamine in an ATP-dependent manner.[2]
 +
 +
Experimental approach:
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1. Integrated sfp into the yeast chromosomal δ-sequences.
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2. Codon-optimize Te bpsA gene for expression in S. cerevisiae and genomically integrated into locus ARS1014a under control of the TDH3 promoter and ADH1 terminator using a previously reported, cloning free Cas9 toolkit.
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3. Use the conventional lithium acetate method to perform transformations.
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4. Use 200 ng pCut_1014a and 500 ng of linear Donor DNA with 500 bp homology to the integration locus ARS1014a.[3]
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===References===
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[1] High-Level Production of the Natural Blue Pigment Indigoidine from Metabolically Engineered Corynebacterium glutamicum for Sustainable Fabric Dyes
 +
 +
[2] Genome-scale metabolic rewiring improves titersrates and yields of the non-native product indigoidine at scale
 +
 +
[3] Production efciency of the bacterial non-ribosomal peptide indigoidine relies on the respiratory metabolic state in S. cerevisiae

Latest revision as of 15:07, 18 September 2022

Svp 4'-Phosphopanthetheinyl-transferase

svp encodes for a 4'-Phosphopanthetheinyl-transferase (PPTase) which transfers a 4'-PPT residue from CoA to a conserved serine residue in the T-Domain of NRPS modules, thus activating these enzymes.

Figure 1: General mechanism of activation the activation of NRP Synthetases from apo- to holo-form.

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
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 173
    Illegal NgoMIV site found at 413
    Illegal NgoMIV site found at 663
    Illegal AgeI site found at 573
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 366

References

  • Takahashi, H., Kumagai, T., Kitani, K., Mori, M., Matoba, Y., & Sugiyama, M. (2007). Cloning and Characterization of a Streptomyces Single Module Type Non-ribosomal Peptide Synthetase Catalyzing a Blue Pigment Synthesis *, 282(12), 9073–9081. doi:10.1074/jbc.M611319200


Information contributed by City of London UK (2021)

Part information is collated here to help future users of the BioBrick registry.

Metadata:

  • Group: City of London UK 2021
  • Author: Janusan Jeyananthan
  • Summary: Added information collated from existing scientific studies

Kinetics:

kcat is 86 min(-1) with S.glaucescens apo-ACP TcmM as the substrate and 11 min(-1) with S.mobaraensis apo-PCP BlmI as the substrate.

For Streptomyces mobaraensis apo-PCP BlmI1: KM=3.9 µM For Streptomyces glaucescens apo-ACP TcmM: KM=3.1 µM

[1]

BBa K302010.png

References

Information contributed by NFLS_Nanjing (2022)

Part information is collated here to help future users of the BioBrick registry.

Metadata:

  • Group: NFLS_Nanjing (2022)
  • Author: Yuming Xue
  • Summary: Added information collated from existing scientific studies

Biology and usage:

1. The gene bpsA is expressed in C. glutamicum and carries strong fluxes toward L-glutamate, a precursor of indigoidine. [1]

2. The gene bpsA catalyzes indigoidine formation from two molecules of glutamine in an ATP-dependent manner.[2]

Experimental approach:

1. Integrated sfp into the yeast chromosomal δ-sequences.

2. Codon-optimize Te bpsA gene for expression in S. cerevisiae and genomically integrated into locus ARS1014a under control of the TDH3 promoter and ADH1 terminator using a previously reported, cloning free Cas9 toolkit.

3. Use the conventional lithium acetate method to perform transformations.

4. Use 200 ng pCut_1014a and 500 ng of linear Donor DNA with 500 bp homology to the integration locus ARS1014a.[3]

References

[1] High-Level Production of the Natural Blue Pigment Indigoidine from Metabolically Engineered Corynebacterium glutamicum for Sustainable Fabric Dyes

[2] Genome-scale metabolic rewiring improves titersrates and yields of the non-native product indigoidine at scale

[3] Production efciency of the bacterial non-ribosomal peptide indigoidine relies on the respiratory metabolic state in S. cerevisiae
  1. ↑ “Svp - 4’-Phosphopantetheinyl Transferase Svp - Streptomyces Mobaraensis - Svp Gene & Protein” n.d. Uniprot. Accessed 12 August, 2021 https://www.uniprot.org/uniprot/Q9F0Q6