Identification of proteins involved in membrane fouling in membrane bioreactors (MBRs) treating municipal wastewater

• Published in: International biodeterioration & biodegradation 2012-11, Vol.75, p.15-22
• Main Authors: Miyoshi, Taro, Aizawa, Tomoyasu, Kimura, Katsuki, Watanabe, Yoshimasa
• Format: Article
• Language: English
• Subjects: amino acid composition; amino acids; bioreactors; Foulant characterization; fouling; gels; Membrane bioreactor; Membrane fouling; municipal wastewater; outer membrane proteins; Proteins; Pseudomonas; sequence analysis; trichloroacetic acid; two-dimensional gel electrophoresis
• ISSN: 0964-8305; 1879-0208
• DOI: 10.1016/j.ibiod.2012.08.003

The proteins that caused membrane fouling in a continuous operation of membrane bioreactors (MBRs) treating real municipal wastewater were investigated in detail. We continuously operated two identical pilot-scale MBRs under different solid retention times (SRTs) and extracted the foulants at the end of the operation. Regardless of the operating conditions, proteins were dominant components in the foulants extracted from the fouled membranes. The extracted proteins were subjected to the separation with two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and the identification through the N-terminal amino acid sequencing analysis. The proteins concentrated by the combination of the crude concentration using an ultra-filtration (UF) membrane and trichloroacetic acid (TCA) precipitation were separated and visualized well on 2D-PAGE gels. The results of 2D-PAGE analysis indicated that the compositions of proteins that caused membrane fouling significantly differed depending on the SRT, although such differences cannot be seen in the amino acid composition analysis. Analyzing selected 2D-PAGE spots by N-terminal amino acid sequencing analysis led to the identification of two well-characterized outer membrane proteins originating from Pseudomonas genus, namely OprF and OprD. To our knowledge, this is the first successful identification of proteins that have caused membrane fouling in continuous operations of MBRs treating real wastewater. ► UF filtration followed by TCA precipitation was an effective concentration method. ► Concentrated proteins were separated well by 2D-PAGE. ► Two proteins that caused membrane fouling were identified as OprF and OprD.