文献类型：期刊文献

中文题名：牡蛎酶解工艺优化及其酶解产物对小鼠睾酮分泌的影响

英文题名：Optimization of Enzymatic Hydrolysis from Crassostrea gigas and Effects of Its Enzymatic Hydrolysate on TM3 Leydig Cells

作者：张雪妍[1];秦小明[1,2,3,4];高加龙[1,2,3,4];林海生[1,2,3,4];章超桦[1,2,3,4];黄艳球[1]

机构：[1]广东海洋大学食品科技学院;[2]广东省水产品加工与安全重点实验室;[3]广东普通高等学校水产品深加工重点实验室;[4]国家贝类加工技术研发分中心(湛江),广东湛江524088

年份：2019

卷号：39

期号：3

起止页码：96

中文期刊名：广东海洋大学学报

外文期刊名：Journal of Guangdong Ocean University

收录：CSTPCD

基金：贝类产业技术体系(CARS-49);广东省应用型科技研发专项资金项目(2016B020235002)

语种：中文

中文关键词：牡蛎;酶解产物;小鼠睾丸间质细胞;增殖活性;睾酮

外文关键词：Crassostrea gigas;enzymatic hydrolysate;leydig cells;proliferative activity;testosterone

中文摘要：【目的】优化太平洋牡蛎(Crassostreagigas)酶解工艺条件,并研究酶解产物及其超滤组分对体外培养的睾丸间质细胞增殖及睾酮分泌的影响。【方法】以多肽得率为参考标准,对木瓜蛋白酶酶解牡蛎的加酶量、初始pH、酶解时间、酶解温度进行单因素及正交实验优化;通过MTT法及酶联免疫吸附试验(ELISA)检测不同浓度牡蛎酶解产物及其超滤组分对TM3小鼠睾丸间质细胞增殖及睾酮分泌的影响。【结果】木瓜蛋白酶最佳酶解条件为加酶量3 000 U/g,初始pH 6. 5,酶解温度62.5℃,酶解时间4.5 h,在此条件下其多肽质量浓度为14.95 mg/mL;牡蛎酶解液及其超滤组分在0.25～1.00 mg/mL质量浓度范围内,与空白对照组相比,能够显著增加TM3细胞的增殖活性(P<0.05),且具有浓度依赖性和时间依赖性。其中,<5 ku和5～10 ku超滤组分细胞增殖活性及促进睾酮分泌活性均最强。【结论】牡蛎酶解产物及其超滤组分可有效增强小鼠睾丸间质细胞增殖活性并促进其分泌睾酮。

外文摘要：【Objective】To optimize the enzymolysis process conditions of Crassostrea gigas and explore the effects of enzymatic hydrolysate and its ultrafiltration fractions on cell proliferation and testosterone secretion in the TM3 leydig cells.【Methods】Based on the peptide yield, the single factor optimization and orthogonal experiments were carried out to get the best amount of enzyme, initial pH, temperature and time of enzymatic hydrolysis for papain to hydrolyze oysters. The MTT assay and ELISA kit were used to detect the effects of the proliferation and secretion of testosterone in the TM3 cells, which were treated by different concentrations of oyster hydrolysate and its ultrafiltration fractions.【Results】The optimal enzymatic hydrolysis conditions for papain are as follows: enzyme addition 3 000 U/g, initial pH 6.5, temperature 62.5 ℃, and time 4.5 h. Its peptide concentration is 14.95 mg/mL. Oyster hydrolysate and its ultrafiltration fraction can significantly increase the proliferation activity of TM3 cells in a concentration range of 0.25-1.00 mg/mL when it is compared with the blank control group in a concentration-and time-dependent manner(P<0.05). The ultrafiltration components of <5 ku and5-10 ku significantly enhanced cell proliferation and promoted testosterone secretion.【Conclusion】The oyster hydrolysate and its ultrafiltration component can effectively enhance the proliferation of mouse Leydig cells and promote the secretion of testosterone.