Rsn‐2‐mediated directed foam enrichment of β‐lactamase

Abstract

Today, the availability of methods for the activity-preserving and cost-efficient downstream processing of enzymes forms a major bottleneck to the use of these valuable tools in technical processes. A promising technology appears to be foam fractionation, which utilizes the adsorption of proteins at a gas-liquid interface. However, the employment of surfactants and the dependency of the applicability on individual properties of the target molecules are considerable drawbacks. Here, we demonstrate that a reversible fusion of the large, surface-active protein Ranaspumin-2 (Rsn-2) to a βlactamase (Bla) enabled both surfactant-free formation of a stable foam and directed enrichment of the enzyme by the foaming. At the same time, Bla maintained 70% of its catalytic activity, which was in stark contrast to the enzyme without fusion to Rsn-2. Rsn-2 predominantly mediated adsorption. Comparable results were obtained after fusion to the structurally more complex penicillin G acylase (PGA) as the target enzyme. The results indicate that using a surface-active protein as a fusion tag might be the clue to the establishment of foam fractionation as a general method for enzyme downstream processing.

Separation and Purification of Recombinant β-Glucosidase with Hydrophobicity and Thermally Responsive Property from Cell Lysis Solution by Foam Separation and Further Purification

“…This indicated that GLEGB cell lysis solution had better foaming property and foam stability than Glu. As can be seen from Figure c, the surface tension of GLEGB decreased faster, and the equilibrium surface tension was lower than that of Glu, indicating that the adsorption capacity of GLEGB at the gas–liquid interface was stronger than that of Glu …”

“…As can be seen from Figure 3c, the surface tension of GLEGB decreased faster, and the equilibrium surface tension was lower than that of Glu, indicating that the adsorption capacity of GLEGB at the gas− liquid interface was stronger than that of Glu. 33 3.1. Foam Property.…”

Foaming and defoaming–concepts and their significance in food and allied industries: a review

“…However, various parameters governing process efficiency need to be optimized and controlled for efficient purification including pH, temperature, water hardness, surface tension, surfactant concentration, etc. [91].…”

Immobilized Phosphoenolpyruvate Carboxylase Beads for Catalytic Carboxylation in Carbon Dioxide-Containing Gas–Liquid Flow

“…Gas–liquid contacting apparatuses such as bubble columns are potentially useful for treating CO 2 -containing gas with biochemical approaches. One of the critical problems associated with the gas–liquid system is the bubbles-induced decrease in the enzyme activity as reported, for example, with respect to lysozyme , and β-lactamase . Effects of gas bubbles on the stability and conformation of enzymes were examined , partly with the purpose of optimizing foam fractionation bioprocesses for effective or selective recovering of proteins of interest.…”

“…One of the critical problems associated with the gas−liquid system is the bubbles-induced decrease in the enzyme activity as reported, for example, with respect to lysozyme 18,19 and β-lactamase. 20 Effects of gas bubbles on the stability and conformation of enzymes were examined 21,22 partly with the purpose of optimizing foam fractionation bioprocesses for effective or selective recovering of proteins of interest. The pH-dependent adsorption of lysozyme to the gas−liquid interface was correlated with the secondary structure of the enzyme.…”