title: Functional display of triphenylmethane reductase for dye removal on the surface of Escherichia coli using N-terminal domain of ice nucleation protein.
abstract: Traditional biological treatment for triphenylmethane dye effluent is stuck with the inaccessibility of dye molecules to intracellular dye-degrading enzyme, thus a high-efficiency and low-cost method for dye decolorization is highly desirable. Here we established a bioremediation approach to display triphenylmethane reductase (TMR) on the surface of Escherichia coli (E. coli) using N-terminal of ice nucleation protein as anchoring motif for triphenylmethane dye decolorization for the first time. Approximately 85% of recombinant protein positioning on the surface of E. coil cells exhibited high activity and stability. The optimal temperature and pH of the surface-displayed TMR are 50 °C and 8.5, respectively. Comparing with other reported microorganisms, the decolorization rate for malachite green of this engineered strain is the highest so far, reaching 640 μmol min(-1) g(-1) dry weight cells. These results indicate that this engineered E. coli strain is a very promising candidate for synthetic dye removal.
authors: Institute of Microbiology, College of Life Science, Zhejiang University, Hangzhou 310058, China.
authors: Fen
authors: F
authors: Gao
authors: Institute of Microbiology, College of Life Science, Zhejiang University, Hangzhou 310058, China. Electronic address: htding@outlook.com.
authors: Haitao
authors: H
authors: Ding
authors: Institute of Microbiology, College of Life Science, Zhejiang University, Hangzhou 310058, China.
authors: Zhuo
authors: Z
authors: Feng
authors: Genclonn Biotech (Hangzhou) Co. Ltd., Hangzhou 310023, China.
authors: Danfeng
authors: D
authors: Liu
authors: Institute of Microbiology, College of Life Science, Zhejiang University, Hangzhou 310058, China. Electronic address: yhzhao225@zju.edu.cn.
authors: Yuhua
authors: Y
authors: Zhao
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry* / Q000378:metabolism*; D001673:Biodegradation, Environmental; D004396:Coloring Agents / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D004926:Escherichia coli / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D060847:Metabolic Engineering; D010088:Oxidoreductases / Q000302:isolation & purification / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D011549:Pseudomonas / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012394:Rosaniline Dyes; D013347:Subcellular Fractions / Q000201:enzymology; D013379:Substrate Specificity; D013696:Temperature; D013997:Time Factors
chemical_list: D001425:Bacterial Outer Membrane Proteins; D004396:Coloring Agents; D011993:Recombinant Fusion Proteins; D012394:Rosaniline Dyes; C063113:ice nucleation protein; C005095:malachite green; D010088:Oxidoreductases; C507510:triphenylmethane reductase, Citrobacter
keywords: Cell surface display; Dye degradation; Ice nucleation protein (INP); Triphenylmethane reductase (TMR)