Part:BBa_K2533055

RBS-ldhA-RBS-lldP-TT

L-lactate permease & D-lactate dehydrogenase

Usage and biology

It encodes L-lactate permease and D-lactate dehydrogenase, which could produce and transport lactate. ldhA could convert pyruvate to D-lactate, lldP could transport lactate out of the cell. In this way, Rhodopseudomonas palustris could effciently transport produced lactate out of the cell, which later will be used by Shewanella.

Characterization

This is one section for lactate production and transportation part.

Figure1:RBS-ldhA-RBS-lldP-TT

DNA Gel Electrophoretic

To make sure that we get the target gene, we did DNA gel electrophoretic for verification. And here is the result.

Figure2:Verification of successful transformation of pSB1C3-RBS-ldhA-RBS-lldP-TT

Our target genes are 2739bp, and as the marker is DS5000, we could be sure that the bright bands in this picture are our target genes.

Electrogenesis

By detecting the production of lactate after expressing protein, we might find out whether RBS-ldhA-RBS-lldP-tt could effectively help Rhodopseudomonas palustris to produce more lactate.

Figure3:shows that our modification is effective. Every gene circuits can help strains produce lactate, and mleS-lldP-ldhA is the most efficient one. Therefore, our construction of gene circuits achieves the goal to help strains produce lactate.

It could be demonstrated that targeted genes could be expressed in the engineered cells. More lactate has been produced by engineered bacteria.