Isolation and Characterization of Collagen from the Body Wall of Sea Cucumber Stichopus monotuberculatus

Abstract

To exploit a new collagen resource from the body wall of tropical sea cucumber, pepsin-solubilized collagen of Stichopus monotuberculatus (PSC-Sm) was isolated and characterized with UV-vis spectra, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), amino acid composition, enzyme-digested peptide maps, Fourier transform infrared spectroscopy (FTIR), maximum transition temperature (Tm ), and solubilities. The maximum absorbance of PSC-Sm was exhibited at 218 nm in UV-vis spectra. The triple helical structure and activity of PSC-Sm could be indicated by FTIR. SDS-PAGE showed that the triple helix of PSC-Sm was formed as (α1 )3 by 3 α1 chain homologous with molecular weight of 137 kDa. The Tm of PSC-Sm and calf skin collagen (CSC) were 30.2 and 35.0 ºC, respectively, which consistent with the result of FTIR that CSC contained more stable triple-helix than PSC-Sm. Peptide maps were different between PSC-Sm and CSC, indicating the differences in their amino acid compositions and sequences. The maximum and minimum solubilities of PSC-Sm were observed at pH 2.0 and 4.0, respectively. A sharp decrease in solubility appeared when NaCl concentration was between 3% and 5%. These results showed that collagen from S. monotuberculatus had the type I collagen characteristics and good thermal stability, and therefore, it could be used as an alternative resource of collagen.

In vitro anti-tyrosinase and anti-elastase activity of collagen from sea cucumber (Holothuria leucospilota)

“…It is a group with specific function to form collagen. The functional groups in sea cucumber (H. leucospilota) are different from other collagens, such as those found in S. variegatus, Oreochromis niloticus, Thunnus albacares, and puffer fish, namely amide A, B, I, II and II (Sionkowska et al, 2015;Zhong et al, 2015).…”

“…The body wall of sea cucumber was soaked in destilled water for 1 h. The dissected body wall of sea cucumber was soaked in 0.1 M NaOH solution for 6 h, then soaked in 0.1 M pH 8.0 Tris-HCl for 24 h, washed using pH 7.0 double destilled water, and then stirred using 0.5 M acetic acid for 48 and 72 h. The filtrates were centrifuged under 8000 rpm speed for 20 min, salted out using 0.9 M NaCl for overnight, and then centrifuged under 8000 rpm speed for 20 min for obtaining deposits. The deposits were dialyzed by using a dialysis bag and 0.1 M acetic acid as medium under a temperature of 4°C for 12 h (Sadowska et al, 2003;Sionkowska et al, 2015;Zhong et al, 2015). Then, the medium was replaced with destilled water, and then dialyzed for a similar period.…”

Kinetic Analysis of the Digestion of Bovine Type I Collagen Telopeptides with Porcine Pepsin

“…ASC was digested with pepsin, as described in the Materials and Methods section. Although bovine type I collagen contains no disulfide bonds and exhibit the same patterns in SDS‐PAGE under reducing and nonreducing conditions (data not shown), we chose reducing conditions according to previous reports (Duan and others ; Zhang and others ; Zhong and others ). Figure A shows the results of a 5% SDS‐PAGE gel run.…”

“…Characterization of the digestion reactions of collagen with various enzymes has been reported (Sato and others ; Morimoto and others ; Kunii and others ). Recently, collagens species isolated from various organisms have been considered for industrial use (Kimura and others ; Nam and others ; Duan and others ; Zeng and others ; Zhang and others , Zhong and others ).…”

Komposisi Kandungan Asam Amino Pada Teripang Emas (Stichoupus horens) di Perairan Pulau Bintan, Kepulauan Riau

“…Kolagen diidentifikasi dan dikarakterisasi mengandung komponen asam amino dengan urutan Gly-X-Y secara berulang (dimana X umumnya proline and Y umumnya hydroxyproline). Kandungan asam amino dari pepsin-solubilized collagen of Stichopus monotuberculatus (PSC-Sm) adalah glisin (32.0%), asam glutamat (12.7%), alanin (9.2%), prolin (8.4%), and hydroxyproline (6.7%) (Zhong et al 2015). Teripang emas memiliki potensi sebagai bahan baku pembuat kolagen karena memiliki kandungan glisin (7,18%), asam glutamat (6,60%), prolin (3,42%), dan alanin (2,53%).…”