Abstract
Gadus morhua eggs contain several nutrients, including polyunsaturated fatty acids, lecithin and glycoproteins. A novel sialoglycopeptide from the eggs of G. morhua (Gm-SGPP) was extracted with 90% phenol and purified by Q Sepharose Fast Flow (QFF) ion exchange chromatography, followed by S-300 gel filtration chromatography. Gm-SGPP contained 63.7% carbohydrate, 16.2% protein and 18.6% N-acetylneuraminic acid. High-performance size exclusion chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) demonstrated that Gm-SGPP is a 7000-Da pure sialoglycopeptide. β-elimination reaction suggested that Gm-SGPP contained N-glycan units. Amino acid N-terminal sequence analysis indicated the presence of Ala-Ser-Asn-Gly-Thr-Gln-Ala-Pro amino acid sequence. Moreover, N-glycan was connected at the third Asn location of the peptide chain through GlcNAc. Gm-SGPP was composed of D-mannose, D-glucuronic acid and D-galactose. Fourier transform-infrared spectroscopy (FT-IR), 1H-nuclear magnetic resonance spectroscopy (1H-NMR) and methylation analysis were performed to reveal the structure profile of Gm-SGPP. In vitro results showed that the proliferation activity of MC3T3-E1 cells was significantly promoted by Gm-SGPP. In vivo data revealed that Gm-SGPP increased the calcium and phosphorus content of tibias and promoted longitudinal bone growth in adolescent rats.
Highlights
Gadidae Gadus morhua is one of the most commercially important fish species, with an annual marine catch of 8 million tons in China
The single peak observed in the High-performance liquid chromatography (HPLC) spectrum indicated that Gm-SGPP was a pure glycopeptide with an estimated molecular weight of 7000 Da (Figure 1C)
A much smaller glycoprotein compared to that isolated from rainbow trout, which has a molecular weight of 260 kDa
Summary
Gadidae Gadus morhua is one of the most commercially important fish species, with an annual marine catch of 8 million tons in China. A nine-carbon monosaccharide derivative, is often located at the terminal monosaccharide of glycoproteins existing in various biological tissues [13,14] and forms an important basis of the glycoprotein/glycopeptide functional diversity [15,16]. Fish eggs contain glycoproteins/glycopeptides, some of which are known to have a high sialic acid content. The structure of sialoglycoprotein in the various fish species is different. We elucidated the structure and biological activity of glycopeptides in. A novel glycopeptide from the eggs of G. morhua (Gm-SGPP) was isolated and purified and its structure profile was identified through a combination of spectroscopic techniques and amino acid sequencing methods. In vitro cell proliferation and in vivo bone growth promotion abilities of the glycopeptide were investigated
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