Secretion of Escherichia coli aminopeptidase P in Bacillus subtilis using the prepro-structure coding region of subtilisin amylosacchariticus

Oyama, Hiroshi; Yoshimoto, Tadashi; Takeshita, Tomoaki; Tsuru, Daisuke

doi:10.1016/0922-338x(89)90032-9

Cited by 7 publications

(1 citation statement)

References 13 publications

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“…Subtilisins are among the most valuable enzymes manufactured by the biotechnology industry (Debabov, 1982;Priest, 19891, yet detailed information concerning the rational design of efficient B. subtilis subtilisin production systems is not generally available. The development of recombinant protein production systems using B. subtilis as the host organism, especially those driven by the subtilisin promoter (Vasantha and Filpula, 1989;Oyama et al, 1989), provides an additional impetus for research in this area. Although subtilisin synthesis is not required for sporulation (Stahl and Ferrari, 19841, its production is triggered (controlled) by mechanisms common to those responsible for the initiation of sporulation (Sonenshein, 1989) and hence it has served as a model for developmentally-associated gene expression.…”

Section: Introductionmentioning

confidence: 99%

Physiological and Genetic Strategies for Enhanced Subtilisin Production by Bacillus subtilis

Pierce

Robertson

Leighton

1992

Biotechnology Progress

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Defined minimal media conditions were used to assess and subsequently enhance the production of subtilisin by genetically characterized Bacillus subtilis strains. Subtilisin production was initiated by the exhaustion or limitation of ammonium in batch and fed-batch cultures. Expression of the subtilisin gene (aprE) was monitored with a chromosomal aprE::lacZ gene fusion. The beta-galactosidase production driven by this fusion reflected subtilisin accumulation in the culture medium. Subtilisin gene expression was temporally extended in sporulation-deficient strains (spoIIG), relative to co-genic sporogenous strains, resulting in enhanced subtilisin production. Ammonium exhaustion not only triggered subtilisin production in asporogenous spoIIG mutants but also shifted carbon metabolism from acetate production to acetate uptake and resulted in the formation of multiple septa in a significant fraction of the cell population. Fed-batch culture techniques, employing the spoIIG strain, were investigated as a means to further extend subtilisin production. The constant provision of ammonium resulted in linear growth, with doubling times of 11 and 36 h in each of two independent experiments. At the lower growth rate, the responses elicited (subtilisin production, glucose metabolism, and morphological changes) during the feeding regime closely approximated the ammonium starvation response, while at the higher growth rate a partial starvation response was observed.

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Section: Introductionmentioning

confidence: 99%