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Welcome to the Biofilm Degradation Team Notebook!

Project Background:

Biofilm production by bacteria is a major concern in ASPs (activated sludge processors). The buildup of these bacteria in these processors inhibits the helpful bacteria from being able to effectively break down several components. Of main concern are the biofilms of bacteria such as Nocardia spp., Thiothrix spp., Sphaerotilus natans, and several others. In order to solve this issue we cloned the genes for alpha Amylase and DispersinB, both which break down the polysaccharide matrix of biofilms, as well as the gene for Aiia, a quorum-sensor blocker, into Nitrosospira multiformis, one of the helpful bacteria in the activated sludge processors. Attached to these genes in their respective chassis is a signaling sequence which dictates the expression of these gene products extracellularly. With the expression of these genes outside the cells there should be a significant decrease in the amount of biofilm buildup by these biofilm-creating bacteria and bacteria such as N. multiformis will be less restricted in breaking down the interested components of ASPs.

Biofilm Team:

Cam Zenger, Jared McOmber, and Jordan Berg

The focus of the biofilm team was to insert the genes for Alpha Amylase, DispersinB, and Aiia with a signaling sequence for extracellular expression into the pSB1C3 chassis and to assay the efficacy of these genes in biofilm reduction in ASPs. Additionally, site-directed mutagenesis was performed on alpha Amylase to remove the PstI restriction site found within the gene.