Extracellular serine-proteinases isolated from Streptomyces alboniger: Partial characterization and effect of aprotinin on cellular structure

Lopes, Angela H.; Coelho, R. R. R.; Meirelles, Maria de Nazareth Leal de; Branquinha, Marta H.; Vermelho, Alane Beatriz

doi:10.1590/s0074-02761999000600010

Cited by 6 publications

(3 citation statements)

References 29 publications

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“…Aprotinin was found to have antibacterial activity by its ability to permeate the cell walls of Gram-positive and Gram-negative bacteria and disintegrate the cytoplasm (Pellegrini, et al , 1992). Andréa Loped et al pointed out the direct correlation between the action of aprotinin and inhibiting growth of the Gram-positive bacterium Streptomyces alboniger (Lopes, et al , 1999). The growth of P. gingivalis and Fusobacterium nucleatum was specifically inhibited by amino-protease inhibitors, such as bestatin (Rogers, et al , 1998, Grenier, et al , 2001).…”

Section: Discussionmentioning

confidence: 99%

“…Accordingly, several investigators have reported that a concentration-dependent relationship exits between protease inhibitor and bacterial growth (Labbe, et al , 2001). Grenier's group showed that there was no inhibition of bacteria with bestatin at 0.02 μg/ml, but when the concentration of bestatin approached 10 μg/ml, the bactericidal function reached a maximum (Grenier, et al , 2001) In the presence of aprotinin, the growth of S. alboniger was also partially or completely inhibited, depending on the concentration of the protease inhibitor (Lopes, et al , 1999). Clearly, then, a higher dose of protease inhibitor has the potential to interfere with the proliferation of bacteria, resulting in an alteration of bacterial composition.…”

Section: Discussionmentioning

confidence: 99%

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Effect of protease inhibitors on the quantitative and qualitative assessment of oral microorganisms

Liu¹,

Saxena²,

Deng³

et al. 2010

FEMS Microbiology Letters

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Protease inhibitor cocktails are routinely added to clinical samples used for proteomic studies to inactivate proteases. Since these same samples are often used for microbial studies, we determined whether addition of protease inhibitors could affect the quantitative or qualitative assessment of microbial profiles. Twenty-two saliva samples were collected and processed immediately with or without the addition of a protease inhibitor cocktail. Conventional cultivation methods were used to evaluate total bacterial growth. Total genomic DNA was isolated and a specific 16S rRNA gene-targeted region was PCR-amplified and separated by denaturing gradient gel electrophoresis. A combination of 1D SDS-PAGE and LC-MS/MS methods was used to determine the effect of the protease inhibitors on the integrity of salivary proteins and peptides. Interestingly, no significant differences were observed in either the bacterial growth and composition or the integrity of salivary proteins between the two groups. Correlation coefficients between the paired samples for total cultivable microbiota (r2=0.847), total mutans streptococci (r2=0.898), total oral lactobacilli (r2=0.933), and total Streptococcus mutans (r2=0.870) also exceeded expected values. The results suggest that the addition of a protease inhibitor cocktail in saliva samples does not impact the growth of oral microbiota or compromise the ability to characterize its composition.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Effect of protease inhibitors on the quantitative and qualitative assessment of oral microorganisms

Liu¹,

Saxena²,

Deng³

et al. 2010

FEMS Microbiology Letters

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“…By using the ammonium sulphate method for protease precipitation, proteases from S. albidoflavus are precipitated by 45% saturation [29] and proteases from S. alboniger, by 40% saturation. [30] After ammonium sulphate precipitation, the pellet was dialyzed and loaded onto a Sephadex G-100 column. The fractions were tested for protease activity and the active fractions (nos.…”

Section: Biotechnology and Biotechnological Equipment 459mentioning

confidence: 99%

Characterization of alkaline protease produced byStreptomyces griseorubensE44G and its possibility for controllingRhizoctoniaroot rot disease of corn

Al-Askar

Rashad

Hafez

et al. 2015

Biotechnology & Biotechnological Equipment

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The antifungal activity of Streptomyces griseorubens E44G against Rhizoctonia solani, the causal agent of root rot disease of corn, was investigated. The mycelial growth of R. solani was inhibited by S. griseorubens E44G, indicating that it has an antifungal potential. The antagonist, S. griseorubens E44G, was detected to have proteolytic activity, using the method of casein hydrolysis. Moreover, the protease production was optimized under submerged conditions. The purification and precipitation of protease were achieved by ammonium sulphate and Sephadex G-100 gel filtration chromatography. Protease activity was detected spectrophotometrically based on the production of tyrosine. The molecular weight of the enzyme (35 kDa) was determined by sodium dodecyl sulphate polyacrylamide gel electrophoresis . The optimum activity of the enzyme was detected at pH 8.5 and 60 C. The results indicated that the enzyme was thermostable and retained full activity even after 1 hour of incubation at 60 C. The purified enzyme substantially inhibited the growth of R. solani, indicating that this enzyme may be actually involved in the antagonistic process.

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Production and partial characterization of extracellular proteinases from Streptomyces malaysiensis, isolated from a Brazilian cerrado soil

et al. 2005

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Streptomyces malaysiensis AMT-3, isolated from a Brazilian cerrado soil, showed proteolytic activities detected by gelatin-sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The optimum proteinase production was obtained when using 2.5% wheat bran and 0.1% yeast extract in the culture medium, after 5 days incubation at 30 degrees C. The enzymatic complex degraded gelatin optimally at pH 7.0, and under these conditions eight proteolytic bands (four serine-proteinases and four metaloproteinases), ranging from 20 to 212 kDa, were detected on the culture supernatant filtrates. In addition, a 35-kDa proteinase was thermostable at 60 degrees C for 120 min. These results point out to the applicability of gelatin zymograms in the characterization of crude enzymatic complexes. According to our results, this enzymatic complex could be used for biotechnological applications.

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Extracellular serine-proteinases isolated from Streptomyces alboniger: Partial characterization and effect of aprotinin on cellular structure

Cited by 6 publications

References 29 publications

Effect of protease inhibitors on the quantitative and qualitative assessment of oral microorganisms

Effect of protease inhibitors on the quantitative and qualitative assessment of oral microorganisms

Characterization of alkaline protease produced byStreptomyces griseorubensE44G and its possibility for controllingRhizoctoniaroot rot disease of corn

Production and partial characterization of extracellular proteinases from Streptomyces malaysiensis, isolated from a Brazilian cerrado soil

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