  2.3.1.236 | more |
constructs harboring mutant aziB, in which site specific mutations are performed for inactivating the KR and DH domains, respectively, are introduced into Streptomyces albus, yielding the recombinant strain AL1007 (to express the KR mutant AziB, G1398A within the NADPH-binding motif GxGxxG), AL1008 (to express the KR mutant AziB, Y1549F at the conserved active site), and AL1009 (to express the DH mutant AziB, H935F within the conserved motif HxxxGxxxxP). Upon HPLC-MS analysis, AL1007, AL1008, and AL1009 fail to produce 5-methyl-NPA, confirming that the reductive and dehydrating actions governed by the KR and DH domains of AziB are essential for the 5-methyl-naphthoic acid formation |
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