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Volume 155, Issue 7

Functional characterization of the first two actinomycete 4-amino-4-deoxychorismate lyase genes Free

Abstract

In some antibiotic producers, aminobenzoic acid (PABA) or its immediate precursor, 4-amino-4-deoxychorismate (ADC), is involved in primary metabolism and antibiotic biosynthesis. In sp. FR-008, a gene putatively encoding a fold-type IV pyridoxal 5′-phosphate (PLP)-dependent enzyme was found within the antibiotic FR-008/candicidin biosynthetic gene cluster, whose inactivation significantly reduced the productivity of antibiotic FR-008 to about 20 % of the wild-type level. Its specific role in PABA formation was further demonstrated by the successful complementation of an mutant. Moreover, a free-standing gene , probably encoding another fold-type IV PLP-dependent enzyme, was cloned from the same strain. Inactivation of reduced antibiotic FR-008 yield to about 57 % of the wild-type level in the mutant, and the complementation of the mutant established its involvement in PABA biosynthesis. Furthermore, a / double mutant only retained about 4 % of the wild-type antibiotic FR-008 productivity, clearly indicating that also contributed to biosynthesis of this antibiotic. Surprisingly, apparently retarded growth of the double mutant was observed on minimal medium, which suggested that both and are involved in PABA biosynthesis for primary metabolism. Finally, both PabC-1 and PabC-2 were shown to be functional ADC lyases by enzymic lysis with the release of pyruvate. and appear to represent the first two functional ADC lyase genes identified in actinomycetes. The involvement of these two ADC lyase genes in both cell growth and antibiotic FR-008 biosynthesis sets an example for the interplay between primary and secondary metabolisms in bacteria.

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Keyword(s): aac(3)IV, apramycin-resistance gene , ADC, 4-amino-4-deoxychorismate , Apr, apramycin , LDH, lactate dehydrogenase , PABA, p-aminobenzoic acid , PermE*, up-mutated promoter of erythromycin-resistance gene , PKS, polyketide synthase and PLP, pyridoxal 5′-phosphate
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2009-07-01
2024-04-16
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Functional characterization of the first two actinomycete 4-amino-4-deoxychorismate lyase genes
Microbiology 155, 2450 (2009); https://doi.org/10.1099/mic.0.026336-0
/content/journal/micro/10.1099/mic.0.026336-0
/content/journal/micro/10.1099/mic.0.026336-0
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