Effect of fruit maturation on N-glycosylation of plant-derived native and recombinant miraculin
Issued Date
2022-05-01
Resource Type
ISSN
09819428
Scopus ID
2-s2.0-85126775358
Pubmed ID
35276597
Journal Title
Plant Physiology and Biochemistry
Volume
178
Start Page
70
End Page
79
Rights Holder(s)
SCOPUS
Bibliographic Citation
Plant Physiology and Biochemistry Vol.178 (2022) , 70-79
Suggested Citation
Kajiura H., Hiwasa-Tanase K., Ezura H., Fujiyama K. Effect of fruit maturation on N-glycosylation of plant-derived native and recombinant miraculin. Plant Physiology and Biochemistry Vol.178 (2022) , 70-79. 79. doi:10.1016/j.plaphy.2022.02.026 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/83254
Title
Effect of fruit maturation on N-glycosylation of plant-derived native and recombinant miraculin
Author(s)
Author's Affiliation
Other Contributor(s)
Abstract
Miracle fruit, Synsepalum dulcificum, produces a unique taste-modifying protein, miraculin (MIR), which has an attractive potential for commercial application as a novel low-calorie sweetener. To establish a stable supply system for MIR, a previous study established a platform for recombinant MIR (rMIR) production in tomato plants and demonstrated that native miraculin from miracle fruit (nMIR) and rMIR were almost identical in their protein modifications with N-glycan. However, neither N-glycosylation nor the influence of fruit maturation on the structural changes of N-glycan have been fully characterized in detail. Here, with a focus on N-glycosylation and the contribution of fruit maturation to N-glycan, we reanalyzed the N-glycosylation of the natural maturation stages of nMIR and rMIR, and then compared the N-glycan structures on MIRs prepared from the fruit at two different maturation stages. The detailed peptide mapping and N-glycosylation analysis of MIRs provided evidence that MIRs have variants, which were derived mainly from the differences in the N-glycan structure in nMIR and the N-glycosylation in rMIR and not from the cleavage of the peptide backbone. N-Glycan analysis of MIRs from the maturation stage of fruits demonstrated that N-glycan structures were similar among nMIRs and rMIRs at every maturation stage. These results indicated that the heterogeneously expressed rMIRs had the same characteristics in post-translational modifications, especially N-glycosylation and N-glycan structures, throughout the maturation stages. This study demonstrated the potential of recombinant protein expressed in tomato plants and paves the way for the commercial use of rMIR.