Partial Characterisation of a Cell Wall Associated Proteinase from Lactobacillus Bulgaricus

Ezzat, N.; Zevaco, C.; Soda, M. El; Gripon, J.C.

doi:10.1007/978-94-009-3733-8_162

Cited by 17 publications

(23 citation statements)

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“…The amount of enzyme recovered from cells decreased with the number of extractions. Such a cascade-like liberation of cellwall proteinases has also been observed in S. cremoris (EXTERKATEand DE VEER, 1985), S. lactis (MONNETet al, 1987) and L. bulgaricus (EZZAT et al, 1987). The amount of activity recovered in the culture medium (120.000 PU) was similar to that extracted from cells probably because of a marked liberation of the enzyme into the medium during culture.…”

Section: Discussionsupporting

confidence: 49%

“…The presence of cell-wall proteinases has been observed in L. helveticus, L. lactis and L. bulgaricus by ARGYLEet al (1976), EZZAT et al (1985EZZAT et al ( , 1987, VESCOVOand BOTTAZZI (1979), as weIl as in L. casei and L. plantarum by EL SODA et al (1986). Little is known about the properties of these proteinases.…”

Section: Introductionmentioning

confidence: 91%

“…Electrophoresis in sodium dodecyl sulfate (SDS)-acrylamide gel showed that the major breakdown product of {3-casein had a slightly lower molecular weight than intact casein, the molecular weight of the other products ranging between 2,500 and 19,000 according to EZZf.T et al (1985). The optimum pH for casein hydrolysis by the enzyme of L. bulgaricus is approximately 6.0 according to ARGYLE et al (1976) and EZZAT et al (1987). MORELLI et al (1986) have detected slow-coagulating variants in L. helveticus whose proteolytic activity is lower than that of fast-coagulating ones.…”

Section: Introductionmentioning

confidence: 99%

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Properties and specificity of a cell-wall proteinase from Lactobacillus helveticus

Zevaco

Gripon

1988

Lait

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SummaryA cell-wall proteinase activity was extracted from cells of Lactobacillus helvetieus grown on MRS medium. The enzyme was characterized after DEAE-Trisacryl chrornatography and gel filtration on Ultrogel AcA 34 column. Two proteolytic fractions of different molecular weights were isolated. Their enzyme properties and specificity were clearly similar indicating that these fractions represented the same enzyme. It is a serine proteinase displaying optimum activity at pH 7.5-8.0 and 42 oc. Peptide bonds cleaved in oes l and {3-casein had no corn mon feature and no clear specificity can be defined. The six main peptides obtained after {3-casein hydrolysis were identified (Arg.-Leug, HisI06-PheI19, LYS'~6-GlnlSZ' ArglsrPhel9o, LeuI9,-Valzo9' TyrI93-VaIZ09) as weil as the two peptides liberated from oes l-casein (Arg.-Ileç, Arg.-Glus). These peptides whose size varies between 6 and 19 residues have probably to be hydrolysed by membrane-bound peptidases prior to their transport through the cytoplasmic membrane.Sorne of those released from the Cterminal part of {3-casein are potentially bitter.

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Section: Discussionsupporting

confidence: 49%

Section: Introductionmentioning

confidence: 91%

Section: Introductionmentioning

confidence: 99%

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Properties and specificity of a cell-wall proteinase from Lactobacillus helveticus

Zevaco

Gripon

1988

Lait

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“…There is little, if any, knowledge about the genetics of the Lactobacillus proteinase system. Cell-wall-bound proteinases have not previously been purified to homogeneity, although partial purification has been reported (Argyle et al, 1976;Ezzat et al, 1987Ezzat et al, , 1988Zevaco & Gripon, 1988). In a previous paper (Nzs et al, 1991) we presented the physiochemical characteristics of the partly purified cell-wall-bound proteinase from Lactobacillus casei NCDO 151.…”

Section: Introductionmentioning

confidence: 99%

Purification and N-terminal amino acid sequence determination of the cell-wall-bound proteinase from Lactobacillus paracasei subsp. paracasei

Naes

Nissen‐Meyer²

1992

Journal of General Microbiology

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The cell-wall-bound proteinase from Lactobacillus paracmei subsp. paracasei NCDO 151 was purified to homogeneity by anion-exchange and hydrophobic-interaction chromatography, chromatofocusing and gelfiltration. The purification resulted in a 40&700-fold increase in specific activity of the proteinase and the final yield was approximately 20 %. Upon chromatofocusing, two proteolytically active components, termed pro135 and proll0, were detected. pro135 had an isoelectric point of 4.2. It had an M, of about 300000 as determined by gelfiltration and 135000 as judged by SDS-PAGE, indicating that it may exist as a dimer in its native state. pro110 had an isoelectric point of 4.4, and an M, of about 150000 as determined by gel-filtration and 110OOO as judged by SDS-PAGE. pro110 appears to be a degradation product of pro135 as they have the same N-terminal amino acid sequence. The first N-terminal amino acid was ambiguous for both components, whereas the sequence from the second to the ninth amino acid was Ala-Lys-Ala-Asn-Ser-Met-Ala-Asn. This is identical to the corresponding sequence of the lactococcal cell-wall-bound proteinases. Although the Lactobacillus proteinase was a little smaller than the lactococcal proteinase, their purification characteristics were very similar, suggesting that these proteinases are related.

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“…bulgaricus strain CNRZ 397 can be released from the cell wall by washing cells with buffer and has been resolved into three peaks by ion exchange chromatography (Ezzat et al 1985(Ezzat et al , 1987. The proteolytic activity of the most active peak was partially characterized.…”

Section: Introductionmentioning

confidence: 99%

Cell-wall-associated proteinase of Lactobacillus delbrueckii subsp. bulgaricus CNRZ 397: differential extraction, purification and properties of the enzyme

Laloi

Atlan

Blanc

et al. 1991

Appl Microbiol Biotechnol

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Whole cells of Lactobacillus delbrueckii subsp. bulgaricus CNRZ 397 were able to hydrolyse alpha- and beta-caseins. Irrespective of the growth medium used, milk or De Man-Rogosa-Sharpe (MRS) broth, identical patterns of alpha- and beta-casein hydrolytic products, respectively, were visualized by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. A soluble proteinase present in cell-wall extracts was active on caseins and displayed the same hydrolytic patterns as whole cells. It was purified from cell-wall extract to homogeneity by ultrafiltration and ion exchange chromatography. The enzyme is a monomer with a molecular mass of 170 kDa, an optimum temperature of 42 degrees C and an optimum pH of 5.5. It was strongly activated by dithiothreitol and partially inhibited by E-64. These properties indicate that cysteine residues play an important role in the enzyme mechanism. The purified proteinase was not able to hydrolyse di- or tripeptides.

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Partial Characterisation of a Cell Wall Associated Proteinase from Lactobacillus Bulgaricus

Cited by 17 publications

References 0 publications

Properties and specificity of a cell-wall proteinase from Lactobacillus helveticus

Properties and specificity of a cell-wall proteinase from Lactobacillus helveticus

Purification and N-terminal amino acid sequence determination of the cell-wall-bound proteinase from Lactobacillus paracasei subsp. paracasei

Cell-wall-associated proteinase of Lactobacillus delbrueckii subsp. bulgaricus CNRZ 397: differential extraction, purification and properties of the enzyme

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