[Construction of an expression vector with elastin-like polypeptide tag to purify xylanase]

Area of Science:

• Biochemistry
• Molecular Biology
• Protein Engineering

Context:

• Recombinant protein purification is crucial for biotechnology and research.
• Traditional purification methods can be time-consuming, expensive, and may damage sensitive proteins.
• Elastin-like polypeptides (ELPs) offer a promising alternative for protein purification due to their unique phase transition properties.

Purpose:

• To report the purification of xylanase using the shortest, sensitive ELP (KV8F-20).
• To design a thermophilic xylanase gene and express it fused to an ELP tag.
• To characterize the efficiency and applicability of ELP-based purification for recombinant proteins.

Summary:

• A thermophilic xylanase gene was fused with an ELP tag via a random coil sequence, creating the pET-22b-SoxB-M2-S-ELP vector.
• Recombinant xylanase was purified using inverse transition cycling and high-speed centrifugation.
• The ELP tag facilitated purification with a 3.16-fold increase in purity, 21.2% recovery, and 64.3% final purity at 22°C.

Impact:

• Elastin-like polypeptide (ELP) serves as a simple, fast, gentle, and economical purification tag for recombinant proteins.
• The developed expression vector is a versatile and dependable tool for purifying various target proteins.
• This method advances efficient and accessible protein purification techniques in molecular biology and biotechnology.