Part:BBa_K4083006

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].

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

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]