TY - JOUR
T1 - Progress toward sourcing plants for new bioconjugation tools
T2 - a screening evaluation of a model peptide ligase using a synthetic precursor
AU - Mahatmanto, Tunjung
AU - Azizah, Isyatul
AU - Buchberger, Alex
AU - Stephanopoulos, Nicholas
N1 - Publisher Copyright:
© 2019, King Abdulaziz City for Science and Technology.
PY - 2019/12/1
Y1 - 2019/12/1
N2 - In the present study, leaves from 39 phylogenetically distant plant species were sampled and screened for asparaginyl endopeptidase ligase activity using mass spectrometry to test the generality of peptide ligases in plants. A modified version of the sunflower trypsin inhibitor-1 precursor was used as the substrate for reactions with leaf crude extracts and protein fractions. Masses consistent with products of asparaginyl endopeptidase activities that cleave and ligate the substrate into cyclic peptide following the reactions were detected in 8 plants: Nerium oleander and Thevetia peruviana of the family Apocynaceae; Bauhinia variegata, Dermatophyllum secundiflorum, Pithecellobium flexicaule, and Prosopis chilensis of the family Fabaceae; Morus alba of the family Moraceae; and Citrus aurantium of the family Rutaceae. This screening result represents a 20% hit rate for finding asparaginyl endopeptidase ligase activity from the arbitrary plants sampled. Analysis following a 2-h reaction of the substrate with the crude extract of D. secundiflorum leaves showed that the yield of cyclic peptide remained stable around 0.5 ± 0.1% of the substrate over the course of the reaction.
AB - In the present study, leaves from 39 phylogenetically distant plant species were sampled and screened for asparaginyl endopeptidase ligase activity using mass spectrometry to test the generality of peptide ligases in plants. A modified version of the sunflower trypsin inhibitor-1 precursor was used as the substrate for reactions with leaf crude extracts and protein fractions. Masses consistent with products of asparaginyl endopeptidase activities that cleave and ligate the substrate into cyclic peptide following the reactions were detected in 8 plants: Nerium oleander and Thevetia peruviana of the family Apocynaceae; Bauhinia variegata, Dermatophyllum secundiflorum, Pithecellobium flexicaule, and Prosopis chilensis of the family Fabaceae; Morus alba of the family Moraceae; and Citrus aurantium of the family Rutaceae. This screening result represents a 20% hit rate for finding asparaginyl endopeptidase ligase activity from the arbitrary plants sampled. Analysis following a 2-h reaction of the substrate with the crude extract of D. secundiflorum leaves showed that the yield of cyclic peptide remained stable around 0.5 ± 0.1% of the substrate over the course of the reaction.
KW - Asparaginyl endopeptidase
KW - Bioconjugation tool
KW - Cyclic peptide
KW - Plant peptide ligase
KW - Protein modification
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U2 - 10.1007/s13205-019-1983-2
DO - 10.1007/s13205-019-1983-2
M3 - Article
AN - SCOPUS:85074691296
SN - 2190-572X
VL - 9
JO - 3 Biotech
JF - 3 Biotech
IS - 12
M1 - 442
ER -