1887

Abstract

Branched-chain fatty acids (BCFAs) typically constitute more than 90 % of the fatty acids of . The authors have previously described two Tn-induced, cold-sensitive, BCFA-deficient (<40 %) mutants ( and ) with lowered membrane fluidity. Sequence analyses revealed that Tn was inserted into different genes of the branched-chain -keto acid dehydrogenase cluster () in these two mutants. The cold-sensitivity and BCFA deficiency of , in which Tn was inserted into , were complemented by cloned . The growth and corresponding BCFA content of the mutants at 37 °C were stimulated by fatty acid precursors bypassing Bkd, 2-methylbutyrate (precursor for odd-numbered anteiso-fatty acids), isobutyrate (precursor for even-numbered iso-fatty acids) and isovalerate (precursor for odd-numbered iso-fatty acids). In contrast, the corresponding Bkd substrates, -ketomethylvalerate, -ketoisovalerate and -ketoisocaproate, exhibited much poorer activity. At 26 °C, 2-methylbutyrate and isovalerate stimulated the growth of the mutants, and at 10 °C, only 2-methylbutyrate stimulated growth. Pyruvate depressed the BCFA content of from 33 % to 27 %, which may be close to the minimum BCFA requirement for . The transcription of was enhanced by Bkd substrates, but not by low temperature. When provided with the BCFA precursors, was able to increase its anteiso-C fatty acid content at 10 °C compared to 37 °C, which is the characteristic response of to low temperature. This implies that Bkd is not the major cold-regulation point of BCFA synthesis.

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2005-02-01
2024-11-11
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