Effect of glucose on sporulation and extracellular amylase production byClostridium perfringens type A in a defined medium

Shih, Neng-Jen; Labbé, Ronald G.

doi:10.1007/bf01570758

Cited by 13 publications

(20 citation statements)

References 26 publications

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“…The sporulation medium we used in these experiments, DSSM, contains ϳ8 mM raffinose. Typically, C. perfringens sporulation media, including DSSM, contain moderate amounts of nutrient-rich ingredients (e.g., proteose peptone and yeast extract) in combination with a slowly utilizable carbohydrate source (e.g., starch or raffinose) (26)(27)(28)(29)45). Therefore, carbohydrate metabolism appears to be an important part of the initiation and/or completion of sporulation by C. perfringens.…”

Section: Discussionmentioning

confidence: 99%

“…Glucose has been shown to act as a catabolite repressor of sporulation in C. perfringens (29,45). The mechanism of catabolite repression of sporulation by glucose in C. perfringens has not been determined, but in Bacillus subtilis, another gram-positive spore-forming bacterium, many catabolite repressor (CR) effects from glucose, including sporulation, have been shown to be mediated by the transcriptional regulator CcpA, a member of the LacI/GalR family of repressor proteins (11,20).…”

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confidence: 99%

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The CcpA Protein Is Necessary for Efficient Sporulation and Enterotoxin Gene (cpe) Regulation inClostridium perfringens

2004

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Clostridium perfringens is the cause of several human diseases, including gas gangrene (clostridial myonecrosis), enteritis necroticans, antibiotic-associated diarrhea, and acute food poisoning. The symptoms of antibiotic-associated diarrhea and acute food poisoning are due to sporulation-dependent production of C. perfringens enterotoxin encoded by the cpe gene. Glucose is a catabolite repressor of sporulation by C. perfringens. In order to identify the mechanism of catabolite repression by glucose, a mutation was introduced into the ccpA gene of C. perfringens by conjugational transfer of a nonreplicating plasmid into C. perfringens, which led to inactivation of the ccpA gene by homologous recombination. CcpA is a transcriptional regulator known to mediate catabolite repression in a number of low-G؉C-content gram-positive bacteria, of which C. perfringens is a member. The ccpA mutant strain sporulated at a 60-fold lower efficiency than the wild-type strain in the absence of glucose. In the presence of 5 mM glucose, sporulation was repressed about 2,000-fold in the wild-type strain and 800-fold in the ccpA mutant strain compared to sporulation levels for the same strains grown in the absence of glucose. Therefore, while CcpA is necessary for efficient sporulation in C. perfringens, glucose-mediated catabolite repression of sporulation is not due to the activity of CcpA. Transcription of the cpe gene was measured in the wild-type and ccpA mutant strains grown in sporulation medium by using a cpe-gusA fusion (gusA is an Escherichia coli gene encoding the enzyme ␤-glucuronidase). In the exponential growth phase, cpe transcription was two times higher in the ccpA mutant strain than in the wild-type strain. Transcription of cpe was highly induced during the entry into stationary phase in wild-type cells but was not induced in the ccpA mutant strain. Glucose repressed cpe transcription in both the wild-type and ccpA mutant strain. Therefore, CcpA appears to act as a repressor of cpe transcription in exponential growth but is required for efficient sporulation and cpe transcription upon entry into stationary phase. CcpA was also required for maximum synthesis of collagenase (kappa toxin) and acted as a repressor of polysaccharide capsule synthesis in the presence of glucose, but it did not regulate synthesis of the phospholipase PLC (alpha toxin).

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

The CcpA Protein Is Necessary for Efficient Sporulation and Enterotoxin Gene (cpe) Regulation inClostridium perfringens

2004

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“…The glucose mediated catabolite repression effect was initially described as the glucose effect (34,35), observed in both gram-positive and gramnegative organisms. Shih and Labbe (36) suggested that glucose mediated catabolite repression of enzyme synthesis is very common in Bacillus sp. There was no significant stimulation of α-amylase synthesis after addition of cyclic AMP in culture medium of Bacillus licheniformis SKB4 as exogenous cAMP could not overcome the repression effect of glucose.…”

Section: Catabolite Repression Of α-Amylase Synthesismentioning

confidence: 99%

Induction and catabolite repression of alpha-amylase synthesis in Bacilluslicheniformis SKB4

Samanta¹,

Jana²,

Kar³

et al. 2017

J Microbiol Biotech Res

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Microbial amylases have an exciting potentiality and are being used extensively in different industries. In this study, regulation of amylase biosynthesis was examined in Bacillus licheniformis SKB4 (wild type) and its mutant strain (8b 2,4-dinitrophenol (uncoupler) at (5mM) concentration. Thus, the pattern of regulation of α-amylase production in the present strain was in multiple forms; it showed the classical glucose effect without stimulation of second messenger system.

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“…Activated cultures were incubated at 37OC for 16-18 h and inoculated (2% inoculum) into media designed for sporulation or amylase production (Shih and LabbC 1994) for 8-10 h. Cell pellets were collected by centrifugation at 10 000 x g, washed twice with 0.05 M phosphate buffer (pH 7.0), resuspended in 0.05 M Tris-HC1 buffer (pH 8.0) containing 1 mM phenylmethylsulfonyl fluoride (PMSF), placed in an ice water bath, and disrupted by sonification at 4OC, 1 minlcycle, until 99% of the cells were disrupted. Cell extracts were collected by centrifugation at 10 000 x g for 60 min.…”

Section: Preparation Of Cell Extractsmentioning

confidence: 99%

“…perfringens (Shih and LabbC 1994). Sporulation in C. perfringens is a nonreversible process and no extracellular amylase accumulates once sporulation is initiated (LabbC and Duncan 1975;Shih and LabbC 1994).…”

Section: Introductionmentioning

confidence: 98%

Characterization and distribution of amylases during vegetative cell growth and sporulation ofClostridium perfringens

Shih¹,

Labbé²

1996

Can. J. Microbiol.

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Clostridium perfringens produced eight extracellular and two intracellular amylolytic activities when examined by zymograms following polyacrylamide gel electrophoresis under native conditions. The major intracellular amylase was isolated from vegetative cells of C. perfringens. It possessed an estimated molecular mass of 112 kDa. Sulfhydryl and phenol functional groups were essential to its activity. The amylase was endo-acting on starch and also hydrolyzed pullulan. Polyclonal antisera against a purified extracellular amylase did not cross-react with intracellular amylase and the two amylases were biochemically different. The distribution of extracellular amylolytic activities of sporulating cells was different from that of vegetative cells, whereas the distribution of intracellular amylolytic activities remained identical. A significant increase of a particular amylase (A8) occurred in the extracellular fluid during sporulation compared with that during vegetative growth. Regulation of the excretion of amylase(s) may be sporulation and enterotoxingenicity related.

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Effect of glucose on sporulation and extracellular amylase production byClostridium perfringens type A in a defined medium

Cited by 13 publications

References 26 publications

The CcpA Protein Is Necessary for Efficient Sporulation and Enterotoxin Gene (cpe) Regulation inClostridium perfringens

The CcpA Protein Is Necessary for Efficient Sporulation and Enterotoxin Gene (cpe) Regulation inClostridium perfringens

Induction and catabolite repression of alpha-amylase synthesis in Bacilluslicheniformis SKB4

Characterization and distribution of amylases during vegetative cell growth and sporulation ofClostridium perfringens

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