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Recombinant Cylindriform Spider silk protein production, purification and characterisation

Bhattacharya, Deep LU (2023) KBKM01 20231
Pure and Applied Biochemistry
Abstract
The global need for recombinant proteins is continually growing. The world protein market was valued at 1.75 billion USD in 2021, with an expected compound annual growth rate (CAGR) of 12.00% between 2022 and 2030. Recombinant proteins have proven indispensable in numerous fields. Their versatility and effectiveness have positioned them as key components in developing innovative therapies, sustainable manufacturing practices, and ground-breaking technologies. Among the diverse array of recombinant proteins, spider silk protein has garnered substantial interest within the scientific community. Its remarkable mechanical properties, such as exceptional strength, elasticity, and biocompatibility, have inspired researchers to explore its... (More)
The global need for recombinant proteins is continually growing. The world protein market was valued at 1.75 billion USD in 2021, with an expected compound annual growth rate (CAGR) of 12.00% between 2022 and 2030. Recombinant proteins have proven indispensable in numerous fields. Their versatility and effectiveness have positioned them as key components in developing innovative therapies, sustainable manufacturing practices, and ground-breaking technologies. Among the diverse array of recombinant proteins, spider silk protein has garnered substantial interest within the scientific community. Its remarkable mechanical properties, such as exceptional strength, elasticity, and biocompatibility, have inspired researchers to explore its potential applications in biomedical engineering to textile manufacturing. However, spiders' aggressive and cannibalistic nature renders their use impractical and limits their scalability, thus making recombinant spider silk protein production the only viable option. To develop a deeper understanding of this protein production pipeline, this Master's thesis aims to analyse and optimise the recombinant production of cylindriform or tubiliform spider silk protein using E.coli Tuner cell line. The study reveals promising results, indicating that low IPTG concentrations (0.001mM) and longer expression times (20 hours) can significantly enhance protein production. Structural modelling using AlphaFold also identifies potential purification issues, providing valuable insights for further investigation. These findings contribute to advancing spider silk protein production, addressing scalability concerns and laying the foundation for future studies on its production. (Less)
Popular Abstract
The increasing global demand for recombinant proteins is driven by their
diverse applications in industries such as biotechnology and medicine. The
global protein market was valued at $1.75 billion in 2021, and it is expected to
grow at a compound annual growth rate (CAGR) of 12.000% between 2022 and
2030. These proteins serve a crucial role in the production of therapeutic
hormones for the treatment of diseases and as industrial biocatalysts.
Spider silk protein is an intriguing protein that has garnered considerable
interest from the scientific community. Silk from spiders possesses
extraordinary qualities, including exceptional tensile strength, elasticity, and
biocompatibility. Due to the aggressive and cannibalistic nature of... (More)
The increasing global demand for recombinant proteins is driven by their
diverse applications in industries such as biotechnology and medicine. The
global protein market was valued at $1.75 billion in 2021, and it is expected to
grow at a compound annual growth rate (CAGR) of 12.000% between 2022 and
2030. These proteins serve a crucial role in the production of therapeutic
hormones for the treatment of diseases and as industrial biocatalysts.
Spider silk protein is an intriguing protein that has garnered considerable
interest from the scientific community. Silk from spiders possesses
extraordinary qualities, including exceptional tensile strength, elasticity, and
biocompatibility. Due to the aggressive and cannibalistic nature of these
creatures, it is impractical to produce this protein through conventional
methods, such as by harvesting it directly from spiders. As an intriguing
alternative, researchers are investigating recombinant production methods in
microorganisms.
The purpose of this Master's thesis is multifaceted. The initial objective is to
produce and purify recombinant cylindriform or tubiliform spider silk using
the E.coli Tuner cell line. This involves cultivating the cells in specially designed
bioreactors and employing protein purification and analysis techniques such as
SDS PAGE (sodium dodecyl sulphate polyacrylamide gel electrophoresis) and
IMAC (immobilised metal affinity chromatography). The second objective is to
characterise the produced silk protein. After the purification procedure,
bioinformatic tools will be utilised to validate the sequences of the purified
protein.
By recombinantly producing spider silk protein, researchers hope to unleash its
potential for a variety of applications, from advanced textile materials to tissue
engineering and drug delivery. This thesis represents a significant step towards
understanding the production and characterization of spider silk protein,
contributing to the field of recombinant protein production and expanding the
potential for innovative applications in a variety of industries. (Less)
Please use this url to cite or link to this publication:
author
Bhattacharya, Deep LU
supervisor
organization
course
KBKM01 20231
year
type
H2 - Master's Degree (Two Years)
subject
keywords
SpiderSilk, Recombinant Protein, Protein production, Protein Purification, Structural Modelling, Applied Biochemistry
language
English
id
9128256
date added to LUP
2023-06-22 11:11:58
date last changed
2023-06-22 11:11:58
@misc{9128256,
  abstract     = {{The global need for recombinant proteins is continually growing. The world protein market was valued at 1.75 billion USD in 2021, with an expected compound annual growth rate (CAGR) of 12.00% between 2022 and 2030. Recombinant proteins have proven indispensable in numerous fields. Their versatility and effectiveness have positioned them as key components in developing innovative therapies, sustainable manufacturing practices, and ground-breaking technologies. Among the diverse array of recombinant proteins, spider silk protein has garnered substantial interest within the scientific community. Its remarkable mechanical properties, such as exceptional strength, elasticity, and biocompatibility, have inspired researchers to explore its potential applications in biomedical engineering to textile manufacturing. However, spiders' aggressive and cannibalistic nature renders their use impractical and limits their scalability, thus making recombinant spider silk protein production the only viable option. To develop a deeper understanding of this protein production pipeline, this Master's thesis aims to analyse and optimise the recombinant production of cylindriform or tubiliform spider silk protein using E.coli Tuner cell line. The study reveals promising results, indicating that low IPTG concentrations (0.001mM) and longer expression times (20 hours) can significantly enhance protein production. Structural modelling using AlphaFold also identifies potential purification issues, providing valuable insights for further investigation. These findings contribute to advancing spider silk protein production, addressing scalability concerns and laying the foundation for future studies on its production.}},
  author       = {{Bhattacharya, Deep}},
  language     = {{eng}},
  note         = {{Student Paper}},
  title        = {{Recombinant Cylindriform Spider silk protein production, purification and characterisation}},
  year         = {{2023}},
}