Coding

Part:BBa_K4083006

Designed by: Arsen Orazbek   Group: iGEM21_NU_Kazakhstan   (2021-10-18)
Revision as of 09:50, 20 October 2021 by Arsenorazbek (Talk | contribs)


rhlB with SacI and SalI sites

The rhlB gene is responsible for the production of rhamnosyltransferase chain B (RhlB) in Pseudomonas aeruginosa.

Usage and Biology

The P. aeruginosa is gram-negative bacillus, and opportunstic pathogen. It secretes rhamnolipids - the rhamnose containing glycolipid biosurfactant. This biosurfactants are used by P. aeruginosa to emulsify the oil sunstances for easy digestion. Thus, rhamnolipids can increase the availability of fats wchich can be important in many different areas like petrleum, bioremediation, cosmetics, food, agroculture, etc. [1] However, due to the toxicity and infectiousness of P. aeruginosa, other alternative organisms are tested. Currently, he organism that showed the best results is Pseudomonas putida.

Our team planned to extract rhL and rhlB genes from P.aeruginosa and to insert them into pRGPDuo2 plasmid obtained from Gauttam, R. [2] We developed the new approach to increase the P. putida's rhamnolpid synthesis by adding nadE gene which encodes NAD synthetase. This way, we hoped to see more rhamnolipid production in engineered P. putida.

The rhamnosyltransferase chain B (RhlB) catalyzes the reaction between 3-(3-hydroxyalkanoyloxy)alkanoic acid and dTDP-L-rhamnose which forms mono-rhamnolipid [1].

800px-BBa_K4083007_rhamnolipid_Metabolic_pathway.png

Part functionality

Reference

[1] Chong, H., & Li, Q. (2017, August 5). Microbial production of rhamnolipids: opportunities, challenges and strategies. Microbial Cell Factories. https://microbialcellfactories.biomedcentral.com/articles/10.1186/s12934-017-0753-2

[2] Gauttam, R., Mukhopadhyay, A., & Singer, S. W. (2020). Construction of a novel dual-inducible duet-expression system for gene (over)expression in Pseudomonas putida. Plasmid, 110. https://doi.org/10.1016/j.plasmid.2020.102514

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 1172
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 65
    Illegal NgoMIV site found at 786
    Illegal NgoMIV site found at 899
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
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