中文摘要：

传统的海参体壁胶原纤维提取方法耗时长,采用简化的胶原纤维提取方法,从水洗次数、NaOH溶液洗涤的时间或次数等方面对海参体壁胶原纤维的提取条件进行优化。结果显示,在海参匀浆与洗涤液比例为1∶10（m∶V）及操作温度为4℃的条件下,去离子水洗涤搅拌3次,每次30min,0.1mol/L pH 8.0Tris—HCl缓冲液（含5mmol/L EDTA,0.5mol/L NaCl）洗涤8h,0.1mol/L NaOH溶液洗涤24h,采用红外分析证实该条件下获得的物质为海参胶原纤维,其提取时间由传统的7d缩短为2d。聚丙烯酰胺凝胶电泳（SDS—PAGE）检测得到清晰条带（116-200kDa）,利用液相色谱—质谱（LC—MS/MS）分析法进行测序得到胶原纤维部分序列（Gly-Leu-Pro-Gly-Ala-Arg-Gly-Ser-Asn-Gly-AsnAsp-Gly-Pro-Ala-Gly-Pro-Arg-Gly-Phe-Asp-Gly-Pro-GluGly-Pro-Arg）,与球海胆（Paracentrotus lividus）I型胶原蛋白前体吻合。采用高效液相色谱法对试验获得的胶原纤维进行氨基酸组成分析,其中甘氨酸含量为32.34%,约占总氨基酸的1/3,脯氨酸和羟脯氨酸含量分别为7.75%,5.10%。

英文摘要：

The traditional extraction method of collagen fibers from sea cucumber body wall takes long time. In this study, the simplified extraction method was adopted to extract collagen fibers from body wall of sea cucumber. The extracting condition was optimized by de- termination of rinsing time or times with deionized water and NaOH solution. The body wall homogenate was mixed with different rins- ing solutions at ratio of 1 ： 10 （m/V）, and the following steps were performed at 4 ℃. The results showed that the collagen fibers was obtained by rinsing with deionized water for 30 min and repeating for 3 times, and then rinsing with 0.1 mol/L Tris--HCl （pH 8.0, containing 0.5 mol/L NaCI and 5 mmol/L EDTA） for 8 h, finally trea- ting with 0.1 mol/L NaOH for 24 h. The collagen fibers of sea cu- cumber body wall was identified by fourier transform infrared spec- troscopy. The extracting time is shortened from 7 days to 2 days. A clear band with molecular mass of 116-200 kDa was detected by sodi- um dodecyl sulfate polyacrylamide gel electrophoresis （ SDS-- PAGE）. The partial sequence of collagen fibers （Gty-Leu-Pro-Gly- Ala -Arg-Gly-Ser-Asn-Gly-Asn-Asp-Gly-Pro-Ala-Gly-Pro-Arg-Gly- Phe-Asp-Gly-Pro-Glu-Gly-Pro-Arg） was detected by liquid chroma- tography-mass spectrometry （LC--MS/MS）, which matched partial alpha collagen type 1 precursor （Paracentrotus lividus）. High performance liquid chromatography （HPLC） analysis showed that the component of glycine in collagen fibers was 32.34%, which accounted for about 1/3 of total amino acid, and proline and hydroxyproline were 7.75% and 5.10%, respectively.