Extracellular enzyme synthesis in the genus Bacillus.

Priest, Fergus G.

doi:10.1128/mmbr.41.3.711-753.1977

Cited by 482 publications

(177 citation statements)

References 201 publications

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“…Bacillus spp. are extensively exploited for protease production [6]. However, the great economic value of protease still gives an impetus to the search for new proteases with novel properties.…”

Section: Introductionmentioning

confidence: 99%

Purification and characterization of alkaline protease from Alcaligenes faecalis

Thangam

Rajkumar

2002

Biotech and App Biochem

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Extracellular alkaline protease from the alkalophilic bacterium Alcaligenes faecalis was purified by a combination of ion-exchange and size-exclusion chromatographic methods, and its properties were examined. The purified enzyme had a specific activity of 563.8 micromol of tyrosine/min per mg of protein and gave a single band on native PAGE and SDS/PAGE with a molecular mass of 67 kDa. Gelatin zymogram also revealed one clear zone of proteolytic activity which corresponded to the band obtained with native PAGE and SDS/PAGE. The enzyme had an optimal pH of 9.0 and exhibited its highest activity at 55 degrees C. The enzyme activity was inhibited by PMSF, suggesting the presence of serine residues at the active site. The enzyme had a K(m) of 1.66 mg/ml and a V(max) of 526 units/min per mg of protein with casein as the substrate.

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Section: Introductionmentioning

confidence: 99%

Purification and characterization of alkaline protease from Alcaligenes faecalis

Thangam

Rajkumar

2002

Biotech and App Biochem

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“…Assuming the membrane forms were precursors of exocellular levansucrase, we propose a two-step mechanism for the secretion process of levansucrase. The number of exoprotein synthesis/secretion sites in a B. suhtilk cell is estimated to 2.5 x lo4.Bacillus suhtilis is known to secrete a large variety of extracellular enzymes [1]. Most of them are synthesized during the stationary phase of growth.…”

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confidence: 99%

“…Bacillus suhtilis is known to secrete a large variety of extracellular enzymes [1]. Most of them are synthesized during the stationary phase of growth.…”

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confidence: 99%

Secretion of Bacillus subtilis levansucrase: a possible two‐step mechanism

Petit‐Glatron

Benyahia

Chambert

1987

European Journal of Biochemistry

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The rate of exocellular levansucrase synthesis in an overproducing (sacUh) strain of Bacillus suhtilis was shown to be directly proportional to the amount of two different transient forms of this enzyme located within the membrane fraction of the cells. The apparent M , of the larger membrane form was 53000, and that of the smaller form 50000; the half-life time of each form was estimated in vivo to be 4-6 s and 32-42 s, respectively. Ethanol treatment of the cells lead to the accumulation of the 53 000-M, form which may represent 1.5% of total membrane proteins. This latter form, partially purified, was transformed in vitro into the 50000-M, form by the action of the Escherichia coli leader peptidase. These enzyme forms were quite different from the exocellular levansucrase since they showed a weak affinity for hydroxyapatite and needed complexed iron to display enzyme activity. Assuming the membrane forms were precursors of exocellular levansucrase, we propose a two-step mechanism for the secretion process of levansucrase. The number of exoprotein synthesis/secretion sites in a B. suhtilk cell is estimated to 2.5 x lo4. [4] the sacUh strain QBl12, when fully induced, produces only exocellular levansucrase (about 8% of total proteins) and, moreover, this enzyme became the unique protein secreted in the culture medium during the exponential phase of growth. Thus, this strain seems to contain optimal characteristics for a biochemical study of levansucrase secretion.The existence of a 53000-M, precursor form of levansucrase was demonstrated by the expression of the cloned structural gene of this enzyme in a minicell-producing strain of Escherichia coli; the nucleotide sequence of the signal peptide was determined [5, 61. The levansucrase precursor form synthesized in minicells in the presence of phenethyl alcohol was found in the outer membrane fraction [7]. However, the yield of the precursor form accumulated in such conditions (less than 0.02% of total protein) was too low for purification. Furthermore, characterization of the other components of a presumed B. subtilis export machinery is not possible using this approach. In order to prepare a large amount of the precursor of exocellular levansucrase, we investigated the conditions of its synthesis and accumulation in

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“…The experiments described do not distinguish between the possibilities that these and other extracellular enzymes are released normally but then readsorbed onto the cell surface, or whether the secretion mechanism itself is inhibited during growth in batch culture. Whichever is correct, the preferential accumulation of glucosyltransferase by continuous cultures of S. mutans is in marked contrast to that of many other extracellular enzymes that are preferentially secreted during the early stationary phase of growth in batch culture [ 13].…”

Section: Polyacrylamide Gel Electrophoresismentioning

confidence: 96%

Production of much larger quantities of extracellular glucosyltransferases by continuous than by batch cultures of Streptococcus mutans

Kenney

1982

FEMS Microbiology Letters

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Extracellular enzyme synthesis in the genus Bacillus.

Cited by 482 publications

References 201 publications

Purification and characterization of alkaline protease from Alcaligenes faecalis

Purification and characterization of alkaline protease from Alcaligenes faecalis

Secretion of Bacillus subtilis levansucrase: a possible two‐step mechanism

Production of much larger quantities of extracellular glucosyltransferases by continuous than by batch cultures of Streptococcus mutans

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