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Studies on multi-functional cosmetics activities of peptide hydrolysates from cultured plant cells of Vigna radiata L.
- Alternative Title
- Studies on multi-functional cosmetics activities of peptide hydrolysates from cultured plant cells of Vigna radiata L.
- Author(s)
- PHAM VAN HIEN
- Alternative Author(s)
- PHAM VAN HIEN
- Advisor
- 변상요
- Department
- 일반대학원 응용생명공학과
- Publisher
- The Graduate School, Ajou University
- Publication Year
- 2015-08
- Language
- eng
- Keyword
- peptide hydrolysate; Mungbean; two dimensional electrophoresis; skin anti-aging; skin-whitening
- Alternative Abstract
- This study was subjected on Mungbean (Vigna radiata L.) peptide hydrolysates that derived from the suspension culture cell. Extraction of proteins from Mungbean powder had been evaluated and gotten the optimal condition consist of using Lysis buffer contained 62.5mM Tris, 10% glycerol and 0.5% SDS and adjust pH to 8. Peptide hydrolysates were prepared via though enzymatic method of combination trypsin and chymotrypsin at ratio of substrate : enzymes as 25:1:1 (w/w/w). Crude peptide hydrolysates suspension was isolated on 40% polyacrylamide gel electrophoresis into the fractions that owned peptides in difference from MW range. The bigger peptides focused on the top of gel and smaller peptides presented on the bottom of gels. Loading peptide standards together with Mungbean peptide hydrolysates and separated on electrophoretic gel, the results found that peptides having MW of around 1000, 500-600 and 200-300 Da focused on fraction 5, fraction 8 and fraction 10, respectively. These results suggested that fraction 5 to 10 focused peptide hydrolysates possessing MW around 1000 Da and less than. Skin anti-aging efficacy of Mungbean peptide hydrolysates was evaluated via though proteomics analysis. Proteins of Normal Human Dermal Fibroblasts treated and non-treated with peptide hydrolysates fractions were analyzed according to 5 mechanisms of skin anti-aging. Results found that Mungbean peptide hydrolysates are the good sources of skin anti-aging. The best efficacy belong on fraction 6 that up-regulated the expression of enzymes of collagen synthesis as P4H1 (111.4%), P4H2 (295.6%), P3H1 (111.7%), P3H2 (106.5%), LH3 (534.9%), PDI (107.5%) and PDIA2 (263.0%); up-regulated the expression of TIMP-1 (859.2%); up-regulated the expression of receptor in ECM-cell linkage as integrin β3 (228.8%); up-regulated the expression of endogenous antioxidant enzymes including GSTA1 (271.5%), GPX2 (178.8%), Catalase (109.1%), GSH (125.7%) and GSTM3 (265.1%). Moreover, Mungbean peptide hydrolysates of fraction 6 down-regulated the expression of MMPs including MMP-1 (33.3%), MMP-2 (61.9%), MMP-3 (49.1%), MMP-8 (32.8%) and MMP-13 (47.6%); down-regulated the expression of cytokines as IL-1β (90.6%), IL-6 (66.5%), NF-κβ (73.2%) and TNF-α (44.4%). Additional, proteomics analysis on proteins of Mouse melanoma B16F10 treated and non-treated with Mungbean peptide hydrolysates fractions observed that Mungbean peptide hydrolysates down-regulated the expression of signal, transfer and enzyme proteins in 4 mechanism of skin whitening. Therefore, these peptides are the good sources for skin whitening. The best efficacy was showed on peptides of fraction 7. Detail, these peptides down-regulated the expression of proteins in transcription of melanogenesis as ACTH-R (54.8%), CREB-1 (19.5%), MITF (35.9%), SCF (15.7%); down-regulated the expression of proteins in post-transcriptional modifications including PKC-β (51.2%), TYRP-2 (9.5%), TYR (90.6%); down-regulated the expression of proteins in sorting of melanosomes and in transfer of melanosomes to melanocyte dendrite tips and to keratinocytes as PMEL (25.6%) and (FOXN1 (43.9%), respectively. Otherwise, the inhibition of melanin production was demonstrated on B16F10 treated with peptide hydrolysates from Mungbean. All of Mungbean peptide fractions inhibited melanin production. The best efficacy was observed on cell treated with peptides of fraction 7 (81.4% of control). These results suggested that Mungbean peptide hydrolysates are the good components for skin whitening.
- Fulltext
- Appears in Collections:
- Graduate School of Ajou University > Department of Applied Biotechnology > 3. Theses(Master)
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