Critical roles of spo0A and spo0H in vegetative alkaline phosphatase production in Bacillus subtilis

Abstract

Growth conditions established to optimize vegetative alkaline phosphatase production and stability in Bacillus subtilis were used to compare alkaline phosphatase synthesis and secretion in isogenic strains JH646 (spoOA12) and JH646MS (spoOA12 abrB15). A mutation in spoOA blocked vegetative alkaline phosphatase production, and a second mutation at the abrB locus resulted in hyperinduction of vegetative alkaline phosphatase. Phosphate regulation of vegetative alkaline phosphatase synthesis was unaffected in the double mutant. spoOH, on a multicopy plasmid, partially overcame the spoOA effect.

Regulation of xylanolytic enzymes in Bacillus subtilis

“…amjE, Nicholson & Chambliss, 1987;bglS, Stulke e t al., 1993;de&, Msadek et al, 1991). An important factor involved in the regulation of many postexponentially expressed genes is the SpoOA protein (Boylan et al, 1988;Hulett & Jensen, 1988;Stulke et al, 1993). It is part of the signalling pathway from a decreased intracellular GTP concentration to the temporal activation of the bglS gene .…”

Bacillus licheniformis APase I gene promoter: a strong well-regulated promoter in B. subtilis

“…However, it is not likely that the hyper-production of APase in B. licheniformis MC14 compared to other Bacillus species is due solely to the existence of multiple APase genes, since it has been shown recently that there are multiple APase genes in B. subtilis . B. subtilis produces (or secretes) only one-sixteenth of the total APase activity produced by B. licheniformis MC 14 under similar culture conditions (Hulett & Jensen, 1988). Therefore, the nature of the hyper-production of APase in B. licheniformis MC14 is more likely to be the result of a difference in the promoter strength of one or more of the APase structural genes.…”

“…5 b, d). The medium (DM) used in these studies is a complete medium in which growth of B. subtilis ceases because of limiting phosphate (Hulett & Jensen, 1988). Induction of the APase I promoter was dependent on the phosphate concentration and was initiated during exponential growth between 3 and 4 h, when the phosphate concentration was 0.1-0.08 mM.…”

“…APase assays of B. subtilis strains were performed on culture supernatants from cells grown in defined medium as described previously Hulett & Jensen, 1988). Units of activity were calculated as pmol p-nitrophenol phosphate hydrolysed min-I at pH 9.5 in 1 M-CHES buffer (Sigma) with 0.1 m~-CoCl,, 70 mM-MgSO, and m~-ZnCl,.…”

“…Efforts to isolate APase structural genes from Bacillus by complementation of an Escherichia coli phoA strain have not been successful (Hulett et al, 1988;Lee et al, 1991). Previous problems in sequencing the APase protein were overcome by employing a new technique using the PVDF (polyvinylidene difluoride) immobilon (Matsudaira, 1987).…”