Recombinant Production of Spider Silk Proteins

Heidebrecht, Aniela; Scheibel, Thomas

doi:10.1016/b978-0-12-407679-2.00004-1

Cited by 115 publications

(117 citation statements)

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“…Secretion is more likely to be successful using yeast or baculovirus expression systems, which have previously been demonstrated to process high molecular weight proteins. 52,89 Both baculovirus and yeast have been used to express silk constructs, but none have yielded similar strengths to that reported for E. coli above. In each of these systems, however, only smaller constructs were analyzed.…”

Section: B Silk Gene Expressionmentioning

confidence: 99%

“…Sequence determination has been hampered by the sheer enormity of the proteins, which have estimated size ranges of 200-350 kDa, 52 with large interspecies size variations due to their repetitive sequence motifs. 53,54 The large number of short repeat regions encompassed within the proteins and their genes represents a formidable challenge for sequencing, precluding standard polymerase chain reaction (PCR) and primer walking strategies, 55 and hampering bioinformatics contig assembly.…”

Section: A Genes and Proteinsmentioning

confidence: 99%

“…Additionally, since the genes are guanine and cytosinerich, depletion of the tRNA pools often occurs before obtaining adequate protein yields. 52 High molecular weight expression, matching that of native silks, appears critical in obtaining high strength synthetic silk. This was demonstrated by Xia et al 85 who produced a range of molecular weight silks in Escherichia coli (E. coli) and found a clear dependence of mechanical properties on molecular weight (see later).…”

Section: B Silk Gene Expressionmentioning

confidence: 99%

“…86 Additionally, unwanted recombination events can lead to shortening of the repetitive genes. 52 One method to improve protein yield and purity is to secrete the protein into the extracellular media, which bypasses the intrinsic production limit of the total amount of protein that can accumulate within a cell. With Gramnegative organisms, such as E. coli, secretion requires crossing of the inner and outer cell membrane, which is a difficult process.…”

Section: B Silk Gene Expressionmentioning

confidence: 99%

“…52 Recent work using transgenic expression of ELPylated spider silk proteins in tobacco, however, appears to have overcome the problem of low molecular weight expression. 92 After purifying the fusion proteins by inverse transition cycling and crosslinking with transglutaminase, high molecular weight protein films could be produced.…”

Section: B Silk Gene Expressionmentioning

confidence: 99%

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With great structure comes great functionality: Understanding and emulating spider silk

et al. 2014

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The overarching aim of biomimetic approaches to materials synthesis is to mimic simultaneously the structure and function of a natural material, in such a way that these functional properties can be systematically tailored and optimized. In the case of synthetic spider silk fibers, to date functionalities have largely focused on mechanical properties. A rapidly expanding body of literature documents this work, building on the emerging knowledge of structure-function relationships in native spider silks, and the spinning processes used to create them. Here, we describe some of the benchmark achievements reported until now, with a focus on the last five years. Progress in protein synthesis, notably the expression on full-size spidroins, has driven substantial improvements in synthetic spider silk performance. Spinning technology, however, lags behind and is a major limiting factor in biomimetic production. We also discuss applications for synthetic silk that primarily capitalize on its nonmechanical attributes, and that exploit the remarkable range of structures that can be formed from a synthetic silk feedstock.

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Section: B Silk Gene Expressionmentioning

confidence: 99%