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Synthetic Spider Silk Production on a Laboratory Scale
Published on: July 18, 2012
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Efficient protein production inspired by how spiders make silk
Nina Kronqvist1, Médoune Sarr1, Anton Lindqvist2
1Division for Neurogeriatrics, Department of NVS, Center for Alzheimer Research, Karolinska Institutet, 141 57 Huddinge, Sweden.
Nature Communications
|May 24, 2017
Summary
Researchers developed a spider silk protein fragment (NT*) to improve the recombinant production of difficult-to-express proteins. This new tool enhances protein solubility and purification, aiding pharmaceutical development and creating synthetic lung surfactant.
Area of Science:
- Biochemistry
- Protein Engineering
- Biotechnology
Background:
- Membrane proteins are crucial pharmaceutical targets but challenging for recombinant production due to aggregation.
- Spider silk proteins achieve high concentrations via micellar structures, with the N-terminal domain (NT) forming a soluble shell.
Purpose of the Study:
- To investigate if fusion to the NT domain can enhance the solubility of non-spidroin proteins.
- To engineer a stabilized, hypersoluble, and pH-insensitive NT mutant (NT*) for improved recombinant protein expression.
Main Methods:
- Designing and constructing a charge-reversed N-terminal domain mutant (NT*).
- Creating fusion proteins between NT* and target proteins, including transmembrane proteins.
- Expressing and purifying fusion proteins in Escherichia coli.
- Testing the efficacy of NT*-based synthetic lung surfactant in an animal model.
Main Results:
- NT*-transmembrane protein fusions yielded up to eight times more soluble protein than conventional tags in E. coli.
- NT* enabled homogeneous purification of transmembrane peptides without chromatography.
- Developed a low-cost synthetic lung surfactant using NT* that demonstrated efficacy in an animal model.
- Showcased efficient expression and purification of other challenging, non-transmembrane proteins.
Conclusions:
- Fusion to the engineered NT* domain significantly enhances the solubility and recombinant production of aggregation-prone proteins, including membrane proteins.
- NT* provides a versatile and efficient tool for purifying difficult proteins and developing novel biomaterials like synthetic lung surfactant.
- This approach offers a valuable new strategy for tackling 'reluctant proteins' in biotechnology and pharmaceutical research.
