Difference between revisions of "Part:BBa K5477035"
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UGT2B15 (UDP-glucuronosyltransferase 2B15) is an enzyme belonging to the UDP-glucuronosyltransferase (UGT) family, which plays a role in the phase II metabolism of xenobiotics and endogenous compounds. UGT2B15 is primarily involved in the process of glucuronidation, a biochemical reaction where glucuronic acid is transferred to lipophilic molecules, making them more water-soluble and easier to excrete from the body. This detoxification process is essential for eliminating harmful substances such as drugs, environmental toxins, and metabolic byproducts. In particular, UGT2B15 is responsible for the glucuronidation of a wide range of compounds, including bisphenol A (BPA), steroid hormones (like androgens), and pharmaceuticals. By conjugating these substances with glucuronic acid, UGT2B15 facilitates their removal via urine or bile, reducing their biological activity and toxicity. The enzyme is expressed predominantly in the liver but is also found in other tissues such as the kidney, prostate, and gastrointestinal tract, where it participates in localized detoxification processes (1).
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This composite part consists of UDP-glucuronosyltransferase 2B15 (UGT2B15) [https://parts.igem.org/Part:BBa_K54770169 BBa_K54770169 ], pGAL1/10 bidirectional promoter [https://parts.igem.org/Part:BBa_K5477005 BBa_K5477005] and UDP-glucose dehydrogenase [https://parts.igem.org/Part:BBa_K5477018 BBa_K5477018]. pGAL1/10 drives the expression of the two enzymes in the opposite direction. UDPD synthesizes the necessary precursor (UDP-glucoronic acid) that UGT2B15 uses for phase II detoxification of BPA (1) (2).  
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By incorporating UGT2B15 into the detoxification system, the goal is to metabolize BPA through glucuronidation, rendering it less harmful and easier to excrete from the cell or organism.  
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The composite part was cloned using the method of USER-cloning into YCp-H. YCp-H is a centromeric plasmid used in yeast that includes a HIS3 marker, allowing for selection in histidine auxotrophic yeast strains. Like other CEN plasmids, YCp-H contains a CEN sequence, ensuring that the plasmid replicates and segregates similarly to yeast chromosomes. This results in a low copy number (typically one to two copies per cell), providing stable maintenance of the plasmid.
  
 
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===References===
 
===References===
  
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1. Ramírez, Viviana & Gálvez Ontiveros, Yolanda & Porras, Patricia & Martinez-Gonzalez, Luis & Rivas, Ana & Alvarez-Cubero, María. (2021). METABOLIC pathways, alterations in MIRNAS expression and effects of genetic polymorphisms of bisphenol a analogues: A SYSTEMATIC review. Environmental Research. 197. 111062. 10.1016/j.envres.2021.111062.
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1. Hanioka N, Naito T, Narimatsu S. Human UDP-glucuronosyltransferase isoforms involved in bisphenol A glucuronidation. Chemosphere. 2008 Dec 1;74(1):33–6.
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2. Oka T, Jigami Y. Reconstruction of de novo pathway for synthesis of UDP-glucuronic acid and UDP-xylose from intrinsic UDP-glucose in Saccharomyces cerevisiae. FEBS J. 2006 Jun;273(12):2645–57.

Revision as of 01:39, 2 October 2024


UDPD-pGAL1/10-UGT2B15 detox module against BPA


This composite part consists of UDP-glucuronosyltransferase 2B15 (UGT2B15) BBa_K54770169 , pGAL1/10 bidirectional promoter BBa_K5477005 and UDP-glucose dehydrogenase BBa_K5477018. pGAL1/10 drives the expression of the two enzymes in the opposite direction. UDPD synthesizes the necessary precursor (UDP-glucoronic acid) that UGT2B15 uses for phase II detoxification of BPA (1) (2).

The composite part was cloned using the method of USER-cloning into YCp-H. YCp-H is a centromeric plasmid used in yeast that includes a HIS3 marker, allowing for selection in histidine auxotrophic yeast strains. Like other CEN plasmids, YCp-H contains a CEN sequence, ensuring that the plasmid replicates and segregates similarly to yeast chromosomes. This results in a low copy number (typically one to two copies per cell), providing stable maintenance of the plasmid.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 3466
    Illegal PstI site found at 542
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 3466
    Illegal PstI site found at 542
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 3466
    Illegal BglII site found at 1122
    Illegal BamHI site found at 3286
    Illegal BamHI site found at 3580
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 3466
    Illegal PstI site found at 542
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 3466
    Illegal PstI site found at 542
    Illegal AgeI site found at 1828
  • 1000
    COMPATIBLE WITH RFC[1000]


References

1. Hanioka N, Naito T, Narimatsu S. Human UDP-glucuronosyltransferase isoforms involved in bisphenol A glucuronidation. Chemosphere. 2008 Dec 1;74(1):33–6.

2. Oka T, Jigami Y. Reconstruction of de novo pathway for synthesis of UDP-glucuronic acid and UDP-xylose from intrinsic UDP-glucose in Saccharomyces cerevisiae. FEBS J. 2006 Jun;273(12):2645–57.