Details
| Original language | English |
|---|---|
| Pages (from-to) | 47-55 |
| Number of pages | 9 |
| Journal | Process biochemistry |
| Volume | 73 |
| Publication status | Published - 27 Jul 2018 |
Abstract
A prolyl endopeptidase, FvpP, was identified in the culture supernatant of the basidiomycete Flammulina velutipes and functionally expressed for gene verification in Aspergillus oryzae. The wild-type FvpP with a molecular mass of 50 kDa under denaturing conditions, exhibited optima at pH 5, and 45 °C and an isoelectric point of 3.8. Five mM Fe2+ and Fe3+ activated the prolyl endopeptidase, while it was not affected by 10 mM CaCl2, EDTA, and Pepstatin A. Five mM Cu2+, Mn2+, Zn2+, Ni2+, Co2+ and low Z-Pro-prolinal concentrations decreased the enzyme activity. Concluded from comparative RP-HPLC and high-resolution QTOF-MS/MS analyses of the hydrolytic cleavage products of gliadin, casein and gelatin, FvpP possessed a P1-P1′-substrate specificity similar to the prolyl endopeptidase from Aspergillus niger (An-Pep), although the sequence similarity was only 39%. FvpP and An-Pep (S28.004; MEROPS) are unique in the serine peptidase family S28, because they share more sequence similarity with Pro-X carboxypeptidase (S28.001; MEROPS) and dipeptidyl peptidase II (S28.002; MEROPS) than with prolyl oligopeptidases of the serine peptidase family S09. As a potential degrader of the antigenic epitopes of α-gliadin, FvpP might provide an efficient tool for the processing of wheat and other gluten containing cereals to better digestible foods for celiac patients.
Keywords
- Aspergillus oryzae, Celiac disease, Flammulina velutipes, Prolyl endopeptidase, S28 family
ASJC Scopus subject areas
- Chemical Engineering(all)
- Bioengineering
- Biochemistry, Genetics and Molecular Biology(all)
- Biochemistry
- Immunology and Microbiology(all)
- Applied Microbiology and Biotechnology
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In: Process biochemistry, Vol. 73, 27.07.2018, p. 47-55.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - A prolyl endopeptidase from Flammulina velutipes for the possible degradation of celiac disease provoking toxic peptides in cereal proteins
AU - Schulz, Kathrin
AU - Giesler, Lucienne
AU - Linke, Diana
AU - Berger, Ralf G.
N1 - Funding information: We thank Ulrich Krings for nLC-ESI-QTOF-MS/MS sequencing. Furthermore, the authors gratefully acknowledge Elizabeth Skellam for providing the pTAex3 vector and the Aspergillus oryzae strain.
PY - 2018/7/27
Y1 - 2018/7/27
N2 - A prolyl endopeptidase, FvpP, was identified in the culture supernatant of the basidiomycete Flammulina velutipes and functionally expressed for gene verification in Aspergillus oryzae. The wild-type FvpP with a molecular mass of 50 kDa under denaturing conditions, exhibited optima at pH 5, and 45 °C and an isoelectric point of 3.8. Five mM Fe2+ and Fe3+ activated the prolyl endopeptidase, while it was not affected by 10 mM CaCl2, EDTA, and Pepstatin A. Five mM Cu2+, Mn2+, Zn2+, Ni2+, Co2+ and low Z-Pro-prolinal concentrations decreased the enzyme activity. Concluded from comparative RP-HPLC and high-resolution QTOF-MS/MS analyses of the hydrolytic cleavage products of gliadin, casein and gelatin, FvpP possessed a P1-P1′-substrate specificity similar to the prolyl endopeptidase from Aspergillus niger (An-Pep), although the sequence similarity was only 39%. FvpP and An-Pep (S28.004; MEROPS) are unique in the serine peptidase family S28, because they share more sequence similarity with Pro-X carboxypeptidase (S28.001; MEROPS) and dipeptidyl peptidase II (S28.002; MEROPS) than with prolyl oligopeptidases of the serine peptidase family S09. As a potential degrader of the antigenic epitopes of α-gliadin, FvpP might provide an efficient tool for the processing of wheat and other gluten containing cereals to better digestible foods for celiac patients.
AB - A prolyl endopeptidase, FvpP, was identified in the culture supernatant of the basidiomycete Flammulina velutipes and functionally expressed for gene verification in Aspergillus oryzae. The wild-type FvpP with a molecular mass of 50 kDa under denaturing conditions, exhibited optima at pH 5, and 45 °C and an isoelectric point of 3.8. Five mM Fe2+ and Fe3+ activated the prolyl endopeptidase, while it was not affected by 10 mM CaCl2, EDTA, and Pepstatin A. Five mM Cu2+, Mn2+, Zn2+, Ni2+, Co2+ and low Z-Pro-prolinal concentrations decreased the enzyme activity. Concluded from comparative RP-HPLC and high-resolution QTOF-MS/MS analyses of the hydrolytic cleavage products of gliadin, casein and gelatin, FvpP possessed a P1-P1′-substrate specificity similar to the prolyl endopeptidase from Aspergillus niger (An-Pep), although the sequence similarity was only 39%. FvpP and An-Pep (S28.004; MEROPS) are unique in the serine peptidase family S28, because they share more sequence similarity with Pro-X carboxypeptidase (S28.001; MEROPS) and dipeptidyl peptidase II (S28.002; MEROPS) than with prolyl oligopeptidases of the serine peptidase family S09. As a potential degrader of the antigenic epitopes of α-gliadin, FvpP might provide an efficient tool for the processing of wheat and other gluten containing cereals to better digestible foods for celiac patients.
KW - Aspergillus oryzae
KW - Celiac disease
KW - Flammulina velutipes
KW - Prolyl endopeptidase
KW - S28 family
UR - http://www.scopus.com/inward/record.url?scp=85050914400&partnerID=8YFLogxK
U2 - 10.1016/j.procbio.2018.07.019
DO - 10.1016/j.procbio.2018.07.019
M3 - Article
AN - SCOPUS:85050914400
VL - 73
SP - 47
EP - 55
JO - Process biochemistry
JF - Process biochemistry
SN - 1359-5113
ER -