Difference between revisions of "Part:BBa K2963043"

Revision as of 13:33, 19 October 2019

Producing γ-PGA with different D/L glutamate monomer ratio(R) (two lacO in Ptac)

Usage and Biology

We used this part to produce different D/L monomer ratios of γ-PGA.

The BCA genes from Bacillus sp. encode a γ-PGA synthetase located on the cell membrane which is capable of polymerizing glutamic acid to form poly-γ-glutamic acid. In Bacillus licheniformis, BCA are called capBCA. We used the B* gene, a mutant of B gene.

The racE gene is derived from Bacillus subtilis and it encodes a racemase which can converts L-glutamate to D-glutamate.

We assembled the capB*CA genes and racE gene which is under the control of tac promoter containing one lacO together to construct this part using plasmid PZM1.

Characterization

We transferred this part into our chassis microorganism Corynebacterium glutamicum using plasmid PZM1. And we using HPLC to detect the D/L monomer ratio of γ-PGA. The result shows as below: Picture1

Picture1 is the L-glutamate monomer ratio in γ-PGA we have produced using capB*CA genes and the result reaches about over 90%.

Picture2

Picture2 is the result of BBa_K2963043. This part contains capB*CA genes and racE gene which is under the control of tac promoter with one operator(lacO). The L-glutamate monomer ratio reaches about 32%.

When we linked the capB*CA with racE gene, the L-glutamate monnmer ratio was changing. This part is working. All the results show that we have achieved the goal of producing different D/L glutamate monomer ratio of γ-PGA preliminary.

Sequence and Features