Difference between revisions of "Part:BBa K3562001"
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AHL-acylase is a acylase that cleaves AHL by hydrolyzing the amide linkage between the acyl side chain and HSL moiety.(Fig.1)
 
AHL-acylase is a acylase that cleaves AHL by hydrolyzing the amide linkage between the acyl side chain and HSL moiety.(Fig.1)
 
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[[Image:AHL-acylase.png|frame|'''Figure 1:'''Mechanism of AHL-acylase]]
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[[Image:AHL-acylase.png|frame|'''Figure 1''' Mechanism of AHL-acylase]]
 
<p>
 
<p>
 
Quorum sensing factor is closely related to bacterial virulence. Enzymes that degrade AHL can control the concentration of AHL to silence the expression of related genes. In our project, we use these enzymes to control the virulence of Pseudomonas aeruginosa referred to existing experimental therapies.
 
Quorum sensing factor is closely related to bacterial virulence. Enzymes that degrade AHL can control the concentration of AHL to silence the expression of related genes. In our project, we use these enzymes to control the virulence of Pseudomonas aeruginosa referred to existing experimental therapies.
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Same as other acylases, AiiO catalyzes the reaction which cleaves the amide bond of AHL and turns it into a free homoserine lactone and a fatty acid. And it locates inside the bacteria cell. As for the substrate specificity, it may have a strong preference for inactivating AHLs with the acyl side chains greater than eight carbons and may exhibit a slight preference for 3-oxo-substituted AHLs, However, it probably can degrade several short-chain AHLs as well. So it may function as a broad-spectrum acylase inside the cell.
 
Same as other acylases, AiiO catalyzes the reaction which cleaves the amide bond of AHL and turns it into a free homoserine lactone and a fatty acid. And it locates inside the bacteria cell. As for the substrate specificity, it may have a strong preference for inactivating AHLs with the acyl side chains greater than eight carbons and may exhibit a slight preference for 3-oxo-substituted AHLs, However, it probably can degrade several short-chain AHLs as well. So it may function as a broad-spectrum acylase inside the cell.
 
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[[Image:AiiO fig2.png|frame|'''Figure 2:'''Reduction of AHL concentration during incubation with ''Ochrobactrum'' sp. 44 cell extracts. a: This analysis was performed in duplicates. The mean value and the standard errors (indicated as ±value) were calculated. b: Percentage values indicate the initial concentration of AHL. c: Percentage values indicate the concentration of remaining AHL.[1]]]
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[[Image:AiiO fig2.png|frame|'''Figure 2''' Reduction of AHL concentration during incubation with ''Ochrobactrum'' sp. 44 cell extracts. a: This analysis was performed in duplicates. The mean value and the standard errors (indicated as ±value) were calculated. b: Percentage values indicate the initial concentration of AHL. c: Percentage values indicate the concentration of remaining AHL.[1]]]
  
 
===Reference===
 
===Reference===
 
[1]Czajkowski R, Krzyzanowska D, Karczewska J, et al., Inactivation of AHLs by Ochrobactrum sp. A44 depends on the activity of a novel class of AHL acylase[J]. Environmental Microbiology Reports, 2011, 3(1):59-68.
 
[1]Czajkowski R, Krzyzanowska D, Karczewska J, et al., Inactivation of AHLs by Ochrobactrum sp. A44 depends on the activity of a novel class of AHL acylase[J]. Environmental Microbiology Reports, 2011, 3(1):59-68.

Revision as of 13:50, 26 October 2020


AiiO (with His-tag)

AHL-acylase with AHLs (Quorum-sensing factor Acyl Homoserine Lactone) degradation activity.

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]


Usage and Biology

AHL-acylase is a acylase that cleaves AHL by hydrolyzing the amide linkage between the acyl side chain and HSL moiety.(Fig.1)

Figure 1 Mechanism of AHL-acylase

Quorum sensing factor is closely related to bacterial virulence. Enzymes that degrade AHL can control the concentration of AHL to silence the expression of related genes. In our project, we use these enzymes to control the virulence of Pseudomonas aeruginosa referred to existing experimental therapies.

Different AHL degrading enzymes have different substrate preferences and degradation activities. We try to select suitable ones from a variety of enzymes.

Same as other acylases, AiiO catalyzes the reaction which cleaves the amide bond of AHL and turns it into a free homoserine lactone and a fatty acid. And it locates inside the bacteria cell. As for the substrate specificity, it may have a strong preference for inactivating AHLs with the acyl side chains greater than eight carbons and may exhibit a slight preference for 3-oxo-substituted AHLs, However, it probably can degrade several short-chain AHLs as well. So it may function as a broad-spectrum acylase inside the cell.

Figure 2 Reduction of AHL concentration during incubation with Ochrobactrum sp. 44 cell extracts. a: This analysis was performed in duplicates. The mean value and the standard errors (indicated as ±value) were calculated. b: Percentage values indicate the initial concentration of AHL. c: Percentage values indicate the concentration of remaining AHL.[1]

Reference

[1]Czajkowski R, Krzyzanowska D, Karczewska J, et al., Inactivation of AHLs by Ochrobactrum sp. A44 depends on the activity of a novel class of AHL acylase[J]. Environmental Microbiology Reports, 2011, 3(1):59-68.