Conquering isoleucine auxotrophy of Escherichia coli BLR(DE3) to recombinantly produce spider silk proteins in minimal media.
Large-scale production of recombinant spider silk proteins is a long-term goal for their industrial use. Therefore, we have recently developed a process for bacterial production. Due to a highly repetitive gene sequence of spider silks, the host strain E. coli BLR(DE3) was employed since it shows no homologue recombination. Although perfectly suited for production in full media, the BLR strain does not grow in cost-effective minimal media, indicating a previously not reported L: -isoleucine auxotrophy. We provide evidence that mutated threonine deaminase is likely responsible for the detected auxotrophy of BLR.
Schmidt
Martin
M
Department Chemie, Technische Universität München, Lichtenbergstrasse 4, 85747, Garching, Germany.
Römer
Lin
L
Strehle
Martin
M
Scheibel
Thomas
T
D000818:Animals; D003470:Culture Media; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D005349:Fibroins / Q000096:biosynthesis; D007532:Isoleucine / Q000378:metabolism; D009154:Mutation; D017354:Point Mutation; D011994:Recombinant Proteins / Q000096:biosynthesis*; D013913:Threonine Dehydratase / Q000737:chemistry* / Q000235:genetics*
D003470:Culture Media; D011994:Recombinant Proteins; C086087:spidroin 1; D007532:Isoleucine; C076815:spidroin 2; D005349:Fibroins; D013913:Threonine Dehydratase