Binding Peptide-Guided Immobilization of Lipases with Significantly Improved Catalytic Performance Using Escherichia coli BL21(DE3) Biofilms as a Platform

Hao Dong, Wenxue Zhang, Qize Xuan, Yao Zhou, Shengmin Zhou, Jiaofang Huang, Ping Wang

Research output: Contribution to journalArticlepeer-review

10 Scopus citations


Developing novel immobilization methods to maximize the catalytic performance of enzymes has been a permanent pursuit of scientific researchers. Engineered Escherichia coli biofilms have attracted great concern as surface display platforms for enzyme immobilization. However, current biological conjugation methods, such as the SpyTag/SpyCatcher tagging pair, that immobilize enzymes onto E. coli biofilms seriously hamper enzymatic performance. Through phage display screening of lipase-binding peptides (LBPs) and co-expression of CsgB (nucleation protein of curli nanofibers) and LBP2-modified CsgA (CsgALBP2, major structural subunit of curli nanofibers) proteins, we developed E. coli BL21::ΔCsgA-CsgB-CsgALBP2 (LBP2-functionalized) biofilms as surface display platforms to maximize the catalytic performance of lipase (Lip181). After immobilization onto LBP2-functionalized biofilm materials, Lip181 showed increased thermostability, pH, and storage stability. Surprisingly, the relative activity of immobilized Lip181 increased from 8.43 to 11.33 U/mg through this immobilization strategy. Furthermore, the highest loading of lipase on LBP2-functionalized biofilm materials reached up to 27.90 mg/g of wet biofilm materials, equivalent to 210.49 mg/g of dry biofilm materials, revealing their potential as a surface with high enzyme loading capacity. Additionally, immobilized Lip181 was used to hydrolyze phthalic acid esters, and the hydrolysis rate against dibutyl phthalate was up to 100%. Thus, LBP2-mediated immobilization of lipases was demonstrated to be far more advantageous than the traditional SpyTag/SpyCatcher strategy in maximizing enzymatic performance, thereby providing a better alternative for enzyme immobilization onto E. coli biofilms.

Original languageEnglish (US)
Pages (from-to)6168-6179
Number of pages12
JournalACS Applied Materials and Interfaces
Issue number5
StatePublished - Feb 10 2021

Bibliographical note

Funding Information:
This work was sponsored by the National Natural Science Foundation of China (21636003, 21672065, 22077032, and 31872728), the International S&T Innovation Cooperation Key Project (2017YFE0129600), the National Science and Technology Major Project of the Ministry of Science and Technology of China (2020YFA0908900), the Natural Science Foundation of Shanghai (19ZR1477100), the Fundamental Research Funds for the Central Universities (22221818014), and the 111 Project (B18022).

Publisher Copyright:
© 2021 American Chemical Society.


  • biofilm
  • immobilization
  • lipase
  • lipase binding peptide
  • phthalic acid esters

PubMed: MeSH publication types

  • Journal Article


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