Coding

Part:BBa_K4083007

Designed by: Arsen Orazbek   Group: iGEM21_NU_Kazakhstan   (2021-10-18)
Revision as of 09:52, 20 October 2021 by Arsenorazbek (Talk | contribs)


rhlA gene with SalI and SacI sites at ends

rhlA gene codes for rhamnosyltransferase chain A (RhlA) 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 A catalyzes the production of 3-(3-hydroxyalkanoyloxy)alkanoic acid (HAA) from β-hydroxydecanoyl-ACP. HAA is the precursor for rhanolipid production which is catalyzed by RhlB to form mono-rhamnolipids [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 BamHI site found at 94
    Illegal BamHI site found at 654
    Illegal XhoI site found at 830
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None