Biofilm Polysaccharide Display Platform: A Natural, Renewable, and Biocompatible Material for Improved Lipase Performance

Abstract

Most of the microorganisms can form biofilms, which makes biofilms an abundant bioresource to be exploited. Due to the limitations of the application of current immobilization methods for biofilms, we developed an immobilization method called the biofilm polysaccharide display (BPD) strategy while maintaining the native biofilm structure and catalytic microenvironment of Clostridium acetobutylicum B3. Lipase Lip181 showed significant improvements in stability after chemical immobilization. For example, immobilized Lip181 retained 74.23% of its original activity after incubation for 14 days, while free Lip181 was totally deactivated. In addition, immobilized Lip181 maintained high residual activity (pH 5.0–11.0), which showed improved resistance to pH changes. Notably, this method did not decrease but slightly increased the relative activity of Lip181 from 6.39 to 6.78 U/mg. Immobilized Lip181 was used to prepare cinnamyl acetate, and it showed a maximum yield of 85.09%. Overall, this biofilm immobilization method may promote the development of biocatalytic and biofilm materials.

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

“…p-Nitrophenyl palmitate (pNPP) was used as a substrate to identify the immobilization of Lip181SpyCatcher. 18 2.3. Screening of Binding Peptides Specific to Lipase, Lip181.…”

“…Heterologous expression and purification of the lipase, Lip181, were described in our previous study (Section 5, Supporting Information). 18 Purified Lip181 was used as a target, and binding peptides specific to Lip181 were screened according to the manufacturer's instructions. Briefly, a phage-displayed peptide library was co-incubated with Lip181; the specifically bound phages were eluted and further amplified to enrich the pool of binding peptides.…”

Current Status and Future Perspectives of Supports and Protocols for Enzyme Immobilization

“…Another way to use peptides in enzyme immobilization would be to combine proteins with peptides, thus performing a specific and oriented binding appropriately [282]. This combination can preserve the enzyme's binding to the support and maintain its biological activity.…”

Bacterial Materials: Applications of Natural and Modified Biofilms

“…Similarly, Dong et al. [ 60 ] made use of a chemical immobilization strategy based on carbodiimide coupling to covalently link an enzyme to the EPS of a Bacillus subtilis biofilm matrix without reducing enzymatic activity. Moreover, in the same study, magnetic nanoparticles were added to the enzyme‐enriched biofilm.…”