• BMB Reports
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• v.44 no.12
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• pp.787-792
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• 2011

We investigated whether silk fibroin peptide derived from the silkworm, Bombyx mori, could inhibit inflammation and enhance the anti-inflammatory activity of Tat-superoxide dismutase (Tat-SOD), which was previously reported to effectively penetrate various cells and tissues and exert anti-oxidative activity in a mouse model of inflammation. Inflammation was induced by topical treatment of mouse ears with 12-O-tetradecanoylphorbol-13-acetate (TPA). Histological, Western blot, and reverse transcription-polymerase chain reaction data demonstrated that silk fibroin peptide or Tat-SOD alone could suppress elevated levels of cyclooxygenase-2, interleukin-6, interleukin-1beta, and tumor necrosis factor-alpha induced by TPA. Moreover, silk fibroin peptide significantly enhanced the anti-inflammatory activity of Tat-SOD, although it had no influence on in vitro and in vivo transduction of Tat-SOD. Silk fibroin peptide exhibited anti-inflammatory activity in a mice model of inflammation. Therefore, silk fibroin peptide alone or in combination with Tat-SOD might be used as a therapeutic agent for various inflammatory diseases.

• International Journal of Industrial Entomology and Biomaterials
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• v.20 no.2
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• pp.87-91
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• 2010

Silk fibroin model peptide, alanine pentamer was synthesized through solid-phase method and modified with poly(ethylene glycol). Nuclear magnetic resonance spectrometry and Fourier-transform infrared spectroscopy showed the conformation of alanine pentamer, $\beta$-sheet structure and random coil conformation were not changed with PEGylation. Differential scanning calorimetry showed that relatively strong exothermic peak around $180^{\circ}C$ by PEGylation. No cytotoxicity of PEGylated pentamer was observed by L929 cell proliferation test.

• International Journal of Industrial Entomology and Biomaterials
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• v.23 no.2
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• pp.201-206
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• 2011

Using proteases, we produced a peptide mixture from fibroin of the silkworm $Bombyx$ $mori$. Mice received D-galactose by subcutaneous injection for 8 weeks to accelerate aging, and then received the peptide mixture orally for 7 weeks. In the mice aged with D-galactose, the coefficient of friction of hair increased significantly up to 1.6-fold, but in the mice subsequently given the peptide mixture, it was normal. Scanning electron microscopy showed improved hair cuticles in the latter mice too. These results indicate that oral administration with peptides from $Bombyx$ fibroin counters the aging of hair cuticles.

• Journal of Microbiology and Biotechnology
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• v.23 no.12
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• pp.1779-1784
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• 2013

Although enzyme-hydrolyzed silk fibroin has been reported to enhance cognitive function before, it has been still unknown which peptides can improve memory. Here we report that amino acid sequences of three novel peptides were identified from fibroin hydrolysate. Fibroin hydrolysate was obtained by hydrolysis with protease after partial hydrolysis with 5M $CaCl_2$. Synthesized peptides derived from these sequences improved scopolamine-induced memory impairments in mice. We confirmed this hydrolysate had effects that improved learning and memory abilities by performing the Rey-Kim test. From this hydrolysate of silk fibroin, amino acid sequences of eight peptides were identified by LC-MS/MS. Three peptides (GAGAGTGSSGFGPY, GAGAGSGAGSGAGAGSGAGAGY, and SGAGSGAGAGSGAGAGSGA) were synthesized to investigate whether they could improve memory. Passive avoidance test and Morris water maze test were performed, and all peptides showed memory-enhancing abilities on scopolamine-induced memory impairments in mice. In this study, we identified three novel peptides that could improve memory, and that silk fibroin hydrolysate was a mixture of various active peptides that could enhance memory.

• KSBB Journal
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• v.25 no.4
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• pp.395-400
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• 2010

Papain hydrolysate of fibroin was found to be mainly composed of several even-numbered peptides that can be produced at a large scale and can be used as a precursor for biological fine-chemicals such as peptide detergents. Thus, the hydrolysate was further modified to synthesize a peptide mixture of glyceryl esters using the identical enzyme for the production of such chemicals. Formation of glyceryl ester of each peptide was confirmed by identifying peaks of the nominal mass shift of +74 Da in mass spectrometry. Analysis of the mass spectra indicated that glyceryl esters of di- and tetra-peptides were the major constituents of the mixture and that alanylglycine was most preferentially esterified. It also suggests that papain prefers dipeptide to tetrapeptide and alanine to serine or tyrosine at $P_2$ position as substrate for glyceryl esterification. The glyceryl esters were recovered using ion exchange resin and the yield of glyceryl esterification recorded was 17.8% by weight.

• Journal of Microbiology and Biotechnology
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• v.22 no.4
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• pp.494-500
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• 2012

Silk fibroin (SF) peptide has been traditionally used as a treatment for flatulence, spasms, and phlegm. In this study, we examined whether SF peptide enhanced the anti-inflammatory effect of PEP-1-FK506 binding protein (PEP-1-FK506BP) through comparing the anti-inflammatory activities of SF peptide and/or PEP-1-FK506BP. In the presence or absence of SF peptide, transduction levels of PEP-1-FK506BP into HaCaT cells and mice skin and anti-inflammatory activities of PEP-1-FK506BP were identified by Western blot and histological analyses. SF peptide alone effectively reduced both mice ear edema and the elevated levels of cyclooxygenase-2, interleukin-6 and $-1{\beta}$, and tumor necrosis factor-${\alpha}$, showing similar anti-inflammatory effect to that of PEP-1-FK506BP. Furthermore, co-treatment with SF peptide and PEP-1-FK506BP exhibited more enhanced anti-inflammatory effects than the samples treated with SF peptides or PEP-1-FK506BP alone, suggesting the possibility that SF peptide and PEP-1-FK506BP might interact with each other. Moreover, the transduction data demonstrated that SF peptide did not affect the transduction of PEP-1-FK506BP into HaCaT cells and mice skin, indicating that the improvement of anti-inflammatory effect of PEP-1-FK506BP was not caused by enhanced transduction of PEP-1-FK506BP. Thus, these results suggest the possibility that co-treatment with SF peptide and PEP-1-FK506BP may be exploited as a useful therapy for various inflammation-related diseases.

• 한국생물공학회:학술대회논문집
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• 2005.04a
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• pp.440-440
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• 2005

• Journal of Sericultural and Entomological Science
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• v.39 no.2
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• pp.197-202
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• 1997

To hydrolyze silk fibroin was treated with NaOH solution on various concentrations and times. And it was examined that the addition effects of NaHSO3 solution on the solubility and colouring of silk fibroin. As obtained results are as follows; The more increasement of concentration and time of NaOH treatment, the more increasement of solubility but solubility was slight above 3% concentration of NaOH. Fibroin yield was decreased above 3% concentration. This was due to formation of peptide or amino acids below molecular weight 3,000. Most of molecular weight distribution came out to be under 3,000 in 2% concentration and 4hrs of NaOH treatment. The more increasement of adding concentration and 4hrs of NaOH treatment. The more increasement of adding concentration of NaHSO3, the more reduction of solubility but white index of powder increased. In the results of FT-IR spectras of silk fibroin powder obtained by various concentrations of NaHSO3 treatment, the absorbent peak at 3,400 cm-1 which was considered as -CH=N- (azomethine) was disappeared by the more addition of NaHSO3. It showed that absorbent peak of $\beta$-fibroin moved into low temperature region and transferred to $\alpha$- and random coil structure through the DSC experiment. In the results of amino acid analysis, alkali hydrolysis reduced the oxy amino contents acid like serine and tyrosine, but increased the glycine content by the more addition of NaHSO3.

• Korean Journal of Pharmacognosy
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• v.33 no.4 s.131
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• pp.312-318
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• 2002

Effect of the acid hydrolysate of silk fibroin on obesity was investigated in obese(057BL/KsJ-db/db) mice. After 8 weeks feeding of 1%(w/w) or 3%(w/w) fibroin hydrolysate, the extents of reduction in body weight were significantly higher than that of obese control. The weight reduction in female mice was higher than that in male mice. Plasma leptin in male mice increased up to 1.8-fold higher level than obese control by feeding hydrolysate. In case of female mice, however, it rather decreased with increased feeding concentration of hydrolysate. From the results of high glycine and serine contents of peptide fractions contained in fibroin hydrolysate, it was inferred that fibroin peptides might affect xylosyltransferase(XT) activity on chondroitin sulfate synthesis causing to change susceptibility of adipocytes to hormones such as insulin followed by the reduced leptin synthesis in female mice. The result of the higher lipolysis in hydrolysate-fed group than obese control indicated that the reduction in body weight was due to the increased lipolytic activities in male and female mice in common.

• Applied Chemistry for Engineering
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• v.31 no.1
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• pp.67-72
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• 2020

Biomaterials are frequently functionalized with Arg-Gly-Asp (RGD) peptides to provide cell adhesion sites. In this study, RGD peptides were enzymatically coupled on to the surface of fibroin microspheres. Papain exhibited a strong preference for dansyl phenylalanine for the peptide formation with fibroin microspheres. Thus, RGD1 peptide was designed to carry cysteine to both sides of the sequence, glycine as a spacer and two residues of phenylalanine at the C-terminal (CRGDCGFF). The enzymatic modification facilitated by an increasing amount of substrate and by the presence of organic solvent, dimethylsulfoxide at 25% (v/v). Microspheres coupled with RGD1, showed a significantly different precipitation property and an increased apparent volume, possibly due to the steric hindrance of RGD peptides on the surface. Transmission electron microscopy also confirmed the presence of cysteine residues in RGD1 coupled on the surface of microspheres stained with gold nanoparticles. RGD1-microspheres significantly facilitated the growth of murine fibroblast 3T3 cells even under non-adhesion culture conditions.