Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 107, Issue 32, Pages 14059-14063Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1003366107
Keywords
metabolic engineering; glycyl-tRNA; silk fiber; Nephila clavipes; spinning
Categories
Funding
- Ministry of Education, Science, and Technology (MEST) through the National Research Foundation (NRF) [2009-0082332]
- MEST through NRF [R32-2008-000-10142-0]
- National Institutes of Health P41 Tissue Engineering Resource Center
- Air Force Office of Scientific Research (AFOSR)
- Ministry of Education, Science & Technology (MoST), Republic of Korea [KIB 1] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
- National Research Foundation of Korea [2009-0082332, R32-2008-000-10142-0, 2010-50205] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
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Spider dragline silk is a remarkably strong fiber that makes it attractive for numerous applications. Much has thus been done to make similar fibers by biomimic spinning of recombinant dragline silk proteins. However, success is limited in part due to the inability to successfully express native-sized recombinant silk proteins (250-320 kDa). Here we show that a 284.9 kDa recombinant protein of the spider Nephila clavipes is produced and spun into a fiber displaying mechanical properties comparable to those of the native silk. The native-sized protein, predominantly rich in glycine (44.9%), was favorably expressed in metabolically engineered Escherichia coli within which the glycyl-tRNA pool was elevated. We also found that the recombinant proteins of lower molecular weight versions yielded inferior fiber properties. The results provide insight into evolution of silk protein size related to mechanical performance, and also clarify why spinning lower molecular weight proteins does not recapitulate the properties of native fibers. Furthermore, the silk expression, purification, and spinning platform established here should be useful for sustainable production of natural quality dragline silk, potentially enabling broader applications.
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