Part:BBa_K2533053

RBS-mleS-RBS-lldP-RBS-ldhA-TT

Production and transport of lactic acid

Usage and biology

ldhA refers to fermentative D-lactate dehydrogenase, NAD-dependent, performing the conversion of pyruvate to D-lactate. In this way, Rhodopseudomonas palustris could produce lactate more efficiently, which brings Shewanella more carbon sources.

Characterization

This is one section for lactate utilization part.

Figure1:RBS-mleS-RBS-lldP-RBS-ldhA-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-mleS-RBS-lldP-RBS-ldhA-TT

Our target genes are 4339bp, 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 ldhA 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.