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Bee pollen peptides as potent tyrosinase inhibitors with anti-melanogenesis effects in murine b16f10 melanoma cells and zebrafish embryos
(2024-12-01)
Sangtanoo, Papassara
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Srimongkol, Piroonporn
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Saisavoey, Tanatorn
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Puthong, Songchan
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Buakeaw, Anumart
One important functional food ingredient today, valued for its health properties and ability to prevent disease, is bee pollen, which comprises a combination of nectar, pollen from plants, and the secretions of bees. In this research, the tyrosinase (TYR) inhibiting abilities of the peptides derived from bee pollen protein hydrolysates are investigated. Various proteases were utilized to generate these peptides, followed by testing at different concentrations. Tyrosinase inhibition activity was detected in all cases, while the hydrolysate drawn from 5.0% w/v neutrase exhibited the best IC<inf>50</inf> value and was thus investigated further via ultrafiltration to separate the active fractions. The highest potential for tyrosinase inhibition was recorded for the fractions below 0.65 kDa. Subsequent purification steps via SEC and RP-HPLC led to the identification of the VDGYPAAGY (named VY-9) peptide via LC-Q-TOF-MS/MS in fraction F<inf>1–2</inf>, known for its non-toxic and hydrophobic characteristics albeit poor water solubility. The synthesized VY-9 peptide demonstrated competitive inhibition, with IC<inf>50</inf> values of 0.55 ± 0.03 µM for mono-phenolase and 2.54 ± 0.06 µM for di-phenolase activities, as confirmed by molecular docking analysis revealing dominant hydrogen bond interactions with TYR. Effective concentrations of 0.2–1.6 µM of VY-9 showed negligible cytotoxicity in B16F10 cells. Melanin synthesis suppression was examined via qRT-PCR, and western blot in MITF, TYR, TRP-1, and TRP-2. Cell death in zebrafish embryos was evaluated in vivo using a toxicity assay which revealed no significant influence from VY-9, while anti-melanogenic effects were observed when the concentration was 4 µM, suggesting bee pollen-derived peptides’ potential in cosmetic and pharmaceutical depigmentation applications.
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Optimization, isolation, identification and molecular mechanisms in B16F10 melanoma cells of a novel tyrosinase inhibitory peptide derived from split gill mushrooms
(2024-10-01)
Arnamwong, Sarinya
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Kuptawach, Kittisak
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Sangtanoo, Papassara
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Srimongkol, Piroonporn
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Saisavoey, Tanatorn
Hyperpigmentation often arises from an imbalance in melanogenesis, primarily due to the overexpression of tyrosinase (TYR). While the inhibition of TYR presents a common approach to skin whitening, it can lead to undesirable side effects. Thus, there is growing interest in safe and natural alternatives for TYR inhibition. Bioactive compounds and peptides sourced from split gill mushrooms hold promise in this regard. This study aims to optimize the conditions for papain-mediated hydrolysis of split gill mushroom protein to inhibit TYR activity, utilizing response surface methodology (RSM) and central composite design (CCD). Optimal conditions were determined at a temperature of 46.70 °C, a hydrolysis time of 217.09 min, and an enzyme-to-substrate ratio (E/S) of 1.1%. Under these conditions, the resulting hydrolysates exhibited significant TYR inhibition, with an IC<inf>50</inf> value of 117.86 μg/mL and a degree of hydrolysis (DH) of 87.97%. Further purification via ultrafiltration and RP-HPLC yielded a peptide, Tyr-Ala-Ser-Ile-Leu-Leu (YASILL or YL-6), identified through LC-Q-TOF-MS/MS, which competitively inhibited TYR. YL-6 demonstrated an IC<inf>50</inf> value of 3.97 mM for mono-phenolase activity and 6.75 mM for di-phenolase activity. Molecular docking analysis revealed hydrogen bonds and hydrophobic interactions between TYR and YL-6. Treatment of B16F10 cells with YL-6 across concentrations ranging from 10-3000 μM showed no cytotoxic effects.The inhibition of melanin synthesis was investigated via qRT-PCR along with Western blot in MITF, TYR, TRP-1, and TRP-2. The results obtained in this research may prove significant in guiding the development of commercially viable cosmetic products to whiten the skin.
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Formulation Development of Serum Beads: Investigating the Influence of Sodium Alginate/Iota Carrageenan Ratio on Bead Characteristics
(2024-05-01)
Woraphokanunt, Yada
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Teeranachaideekul, Veerawat
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Rittiboon, Aree
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Jatupornpipat, Marisa
Background and Objectives: To develop cosmetic products by using Ionic gelation techniques as the main technique in the production of serum beads. The purpose of this study is to develop the serum beads recipe by studying the ratio of various types of gelling agents for complete serum beads products. Methodology: Test the ratio of gelling agents in serums that create beads with ionic gelation techniques by Sodium Alginate and Iota-carrageenan which affect the appearance and beads texture after forming. Then compare the concentration of the gelling agent used in the suspension of serum beads that affects the ability to suspend serum beads. In this study, Carbopol® ultrez 20 was selected as the type of gelling agent. Main Results: The most suitable ratio for forming serum beads between Sodium Alginate and Iota-carrageenan is 0.20:0.30. Because the appearance of serum beads is spherical which can be pressed, cracked and spread on the surface easily-moderately, which is the most suitable characteristic for serum beads compared with the other ratios. When the serum beads were suspended inside the Carbopol® ultrez 20 gel at different concentrations and sensory tests, it was found that Carbopol® ultrez 20 at a concentration of 0.70 percent was the most favored. After storing in accelerated conditions to test the stability of the product, it was found that all concentrations of the gelling agent Carbopol® ultrez 20 could make the serum beads suspended and disperse inside the gel, but the viscosity was significantly reduced in each formulation. Conclusions: Conclusion: The ratio of Sodium Alginate and Iota-carrageenan affects the formation and hardness of beads. The most suitable ratio in this study is 0.20:0.30 and the gelling agent Carbopol® ultrez 20 at a concentration of 0. 70 percent is the most suitable choice for using as a suspension gel in the serum bead cosmetic formula. Because it is the most stable and has the least change when stored in accelerated condition for 8 weeks.
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Optimization of the phosphate-solubilizing fungus, Aspergillus japonicus SA22P3406, in solid-state cultivation by response surface methodology
(2009-12-01)
Nopparat, Chalit
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Jatupornpipat, Marisa
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Rittiboon, Aree
Phosphorus (P) is one of the essential macronutrients for plant growth and reproduction. Plants acquire P from the soil solution as phosphate anions. However, phosphate anions are extremely reactive and may be immobilized through precipitation with cations such as Ca2+, Mg2+, Fe 3+ and Al3+, depending on the particular properties of the soil and as a result, the phosphate is highly insoluble and unavailable to plants. Application of phosphate-solubilizing fungi (PSF) has been added as fertilizer to increase P uptake and plant growth. The aim of this study was to use response surface methodology to determine the optimal factors of the phosphate-solubilizing fungus Aspergillus japonicus SA22P3406 for use in solid state cultivation, specifically the pH, temperature and ratio of water to dry rice bran (RWB). A. japonicus SA22P3406 was screened from 24 soil samples. Primary screening was carried out by a 10-fold serial dilution of the soil samples at 10-4 to 10-6 and then spread on Pikovskaya agar supplemented with 1.0% w/v of tricalcium phosphate at 37°C for 3-5 d. One hundred and fifty five fungal isolates produced high ratios of clear zone to colony on Pikovskaya agar supplemented with 0.003% w/v of rose bengal at 37°C for 7 d. The ratios were calculated by dividing the area of the clear zone by the area of the colony. Secondary screening was performed by measuring the available phosphate in liquid medium from the 62 fungal strains by CRD and DMRT at 5% probability. Two fungal strains, A. niger SA07P3332 and A. japonicus SA22P3406, were selected as most suitable for solubilizing tricalcium phosphate and producing the highest available phosphate levels of 3.01 and 2.99 mg P <inf>2</inf>O<inf>5</inf> ml-1, respectively. A. japonicus SA22P3406 was cultivated in 5.0 g rice bran to study the optimal conditions using response surface methodology with a central composite design. The optimum conditions of pH, temperature and RWB were 7.37, 36.0°C and 82.10% v/w of dry rice bran, respectively. These conditions were used in the solid-state cultivation and a sporulation curve was determined. The fungal sporulation had a lower than average decadic logarithm of practical quantitative sporulation of 8.46 compared with the estimated quantitative sporulation of 8.53 at the 5th day of incubation.
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The effect of kefir starter on Thai fermented sausage product
(2007-01-01)
Jatupornpipat, Marisa
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Keatikumjorn, Payom
The effect of kefir starter from Wilderness Family Naturals Company on the initial formulation of Thai fermented sausage were evaluated. The differences found among batches in the main microbial populations and pH were not significant. Only, the total acid of batch D (added the kefir starter 15 ml) was significantly higher (P<0.05) than those of batches C, A and B. (control: not added the kefir starter, added the kefir starter 1 and 7 ml. respectively). The odour, flavor and overall acceptability of the B batch was different from those of the other three (P<0.05), C, A and D, which were very similar to each other (P>0.05). It is concluded that the addition of kefir starter (7 ml) could be useful to improve the final quality of Thai fermented sausages. The addition of kefir starter that initiates rapid acidification of the raw meat and that leads to a desirable sensory quality of the end-product are used for the production of fermented sausages, and represents a way of improving and optimizing the sausage fermentation process and achieving tastier, safer, and healthier products.

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