Activity of Fe-S cluster requiring proteins

摘要

The present invention is related to a recombinant host cell, in particular a yeast cell, comprising a dihydroxy-acid dehydratase polypeptide. The invention is also related to a recombinant host cell having increased specific activity of the dihydroxy-acid dehydratase polypeptide as a result of increased expression of the polypeptide, modulation of the Fe—S cluster biosynthesis of the cell, or a combination thereof. The present invention also includes methods of using the host cells, as well as, methods for identifying polypeptides that increase the flux in an Fe—S cluster biosynthesis pathway in a host cell.

主权项

1. A recombinant yeast host cell comprising (i) at least one heterologous polynucleotide encoding a dihydroxy-acid dehydratase (DHAD), and (ii) at least one deletion in an endogenous gene encoding Fe Repressor of Activation-2 (FRA2), wherein the deletion eliminates FRA2 activity,wherein the DHAD has an amino acid sequence that matches a Profile Hidden Markov Model (HMM) prepared using the amino acid sequences of SEQ ID NOs: 164, 168, 230, 232, 298, 310, 344, and 346 with an E value of <10−5, and wherein the polypeptide further comprises all three conserved cysteines, corresponding to positions 56, 129 and 201 in the amino acid sequence of Streptococcus mutans DHAD corresponding to SEQ ID NO: 168.