Lipases have been widely applied in a variety of industrial fields,such as food,pharmaceuticals,biofuels,and biotechnology.Recent years have witnessed a great interest in modifying lipids for the production of triacyl...Lipases have been widely applied in a variety of industrial fields,such as food,pharmaceuticals,biofuels,and biotechnology.Recent years have witnessed a great interest in modifying lipids for the production of triacylglycerols enriched with n-3 polyunsaturated fatty acids(PUFAs).Here,a novel salt-tolerant,organic solvent-stable,and bile salt-activated lipase was purified from golden pompano(Trachinotus ovatus)viscera,which was named as golden pompano lipase(GPL).GPL had a specific activity of 57.2U mg^(-1)with an estimated molecular weight of 14 k Da,exhibited optimal activity at 40℃a nd pH 8.0,and showed K_(m)and V_(max)of 40.16μmol L^(-1)and 769.23μmol L^(-1)min^(-1),respectively.GPL activity was enhanced by Mn^(2+)and sodium deoxycholate.It was active in organic solvents,including methanol,ethanol,chloroform,and hexane.GPL also showed a good salinity tolerance of up to 1 mol L^(-1).n-3PUFA enrichment in the glyceride fraction of golden pompano oil was performed by GPL-catalyzed hydrolysis and yielded a total PUFA concentration of 56.99%.EPA,DHA,and DPA were enriched by 10.4-,3.2-,and 1.8-fold of their initial levels,respectively.This study recognized the industrial applicability of GPL to prepare enriched C_(20-22)n-3 PUFA.展开更多
为提高金鲳鱼加工副产物——鱼皮的利用率,本研究在前期单因素实验的基础上,以加酶量、液料比和提取时间三个单因素建立模型,采用响应面法Box-Behnken设计实验,优化了胃蛋白酶法提取金鲳鱼鱼皮酶溶性胶原蛋白(PSC)的提取工艺,并与酸溶...为提高金鲳鱼加工副产物——鱼皮的利用率,本研究在前期单因素实验的基础上,以加酶量、液料比和提取时间三个单因素建立模型,采用响应面法Box-Behnken设计实验,优化了胃蛋白酶法提取金鲳鱼鱼皮酶溶性胶原蛋白(PSC)的提取工艺,并与酸溶性胶原蛋白(ASC)进行聚丙烯酰胺凝胶电泳、傅里叶变换红外光谱和差示扫描量热仪等基本理化特性的对比研究。结果表明,金鲳鱼鱼皮胶原蛋白的最佳提取工艺为:加酶量1.5%、液料比23∶1 m L/g、酶解时间27.5 h。此条件下通过实验验证得出,金鲳鱼鱼皮胶原蛋白实际提取率为64.68%,与响应面模型理论预测值65.17%基本一致;此外,本实验提取的PSC有较高热变性温度(30.44℃),与金鲳鱼皮ASC有相似的电泳性质和红外峰型,初步鉴定为I型胶原蛋白。该实验为金鲳鱼鱼皮酶溶性胶原的提取提供了理论参考。展开更多
【目的】卵形鲳鲹(Trachinotus ovatus)是广西重要的海水经济鱼类,但近年来随着养殖规模扩大,各种病害侵染引起的疾病频繁暴发,给广西卵形鲳鲹养殖产业造成重大损失。建立卵形鲳鲹小脑来源的细胞系(Cell line from cerebellum of Trachi...【目的】卵形鲳鲹(Trachinotus ovatus)是广西重要的海水经济鱼类,但近年来随着养殖规模扩大,各种病害侵染引起的疾病频繁暴发,给广西卵形鲳鲹养殖产业造成重大损失。建立卵形鲳鲹小脑来源的细胞系(Cell line from cerebellum of Trachinotus ovatus,TOCC),将有利于广西卵形鲳鲹养殖中病毒性病害的分离、鉴定和病害的侵染致病分子机理的研究,以及环境污染和变化的动态监测。【方法】采用胰蛋白酶消化法分离得到卵形鲳鲹小脑组织的原代细胞,并进行原代培养。待原代细胞铺满单层后进行传代培养。对稳定传代的TOCC进行18SrDNA基因测序分析,然后进行生长速率分析和染色体分析,并开展病原菌(溶藻弧菌Vibrio alginolyticus)胞外产物对TOCC的毒性分析。【结果】稳定传代的TOCC为成纤维样细胞,18SrDNA基因分析结果显示其来源于卵形鲳鲹;染色体核型分析发现TOCC具有二倍体特征,特征染色体数目为62条;不同培养条件下TOCC细胞的生长速率不同,28℃生长速度最快;病原菌胞外产物对TOCC具有明显的毒性。【结论】本研究成功建立广西卵形鲳鲹小脑组织来源的细胞系(TOCC)。细菌胞外产物对TOCC细胞毒性结果表明,该细胞系在一定范围内能够替代活体检测,实现体外细胞水平对广西海水养殖中病害侵染和环境污染物的快速检测,为养殖病害的有效预防提供技术支持。展开更多
基金This work was supported by the National Key R&D Programs of China(No.2018YFD0901103)the Program of the Hainan Association for Science and Technology Plans to Youth R&D Innovation(No.QCXM202003)the Hainan Provincial Natural Science Foundation of China(No.2019RC093).
文摘Lipases have been widely applied in a variety of industrial fields,such as food,pharmaceuticals,biofuels,and biotechnology.Recent years have witnessed a great interest in modifying lipids for the production of triacylglycerols enriched with n-3 polyunsaturated fatty acids(PUFAs).Here,a novel salt-tolerant,organic solvent-stable,and bile salt-activated lipase was purified from golden pompano(Trachinotus ovatus)viscera,which was named as golden pompano lipase(GPL).GPL had a specific activity of 57.2U mg^(-1)with an estimated molecular weight of 14 k Da,exhibited optimal activity at 40℃a nd pH 8.0,and showed K_(m)and V_(max)of 40.16μmol L^(-1)and 769.23μmol L^(-1)min^(-1),respectively.GPL activity was enhanced by Mn^(2+)and sodium deoxycholate.It was active in organic solvents,including methanol,ethanol,chloroform,and hexane.GPL also showed a good salinity tolerance of up to 1 mol L^(-1).n-3PUFA enrichment in the glyceride fraction of golden pompano oil was performed by GPL-catalyzed hydrolysis and yielded a total PUFA concentration of 56.99%.EPA,DHA,and DPA were enriched by 10.4-,3.2-,and 1.8-fold of their initial levels,respectively.This study recognized the industrial applicability of GPL to prepare enriched C_(20-22)n-3 PUFA.
文摘为提高金鲳鱼加工副产物——鱼皮的利用率,本研究在前期单因素实验的基础上,以加酶量、液料比和提取时间三个单因素建立模型,采用响应面法Box-Behnken设计实验,优化了胃蛋白酶法提取金鲳鱼鱼皮酶溶性胶原蛋白(PSC)的提取工艺,并与酸溶性胶原蛋白(ASC)进行聚丙烯酰胺凝胶电泳、傅里叶变换红外光谱和差示扫描量热仪等基本理化特性的对比研究。结果表明,金鲳鱼鱼皮胶原蛋白的最佳提取工艺为:加酶量1.5%、液料比23∶1 m L/g、酶解时间27.5 h。此条件下通过实验验证得出,金鲳鱼鱼皮胶原蛋白实际提取率为64.68%,与响应面模型理论预测值65.17%基本一致;此外,本实验提取的PSC有较高热变性温度(30.44℃),与金鲳鱼皮ASC有相似的电泳性质和红外峰型,初步鉴定为I型胶原蛋白。该实验为金鲳鱼鱼皮酶溶性胶原的提取提供了理论参考。
文摘【目的】卵形鲳鲹(Trachinotus ovatus)是广西重要的海水经济鱼类,但近年来随着养殖规模扩大,各种病害侵染引起的疾病频繁暴发,给广西卵形鲳鲹养殖产业造成重大损失。建立卵形鲳鲹小脑来源的细胞系(Cell line from cerebellum of Trachinotus ovatus,TOCC),将有利于广西卵形鲳鲹养殖中病毒性病害的分离、鉴定和病害的侵染致病分子机理的研究,以及环境污染和变化的动态监测。【方法】采用胰蛋白酶消化法分离得到卵形鲳鲹小脑组织的原代细胞,并进行原代培养。待原代细胞铺满单层后进行传代培养。对稳定传代的TOCC进行18SrDNA基因测序分析,然后进行生长速率分析和染色体分析,并开展病原菌(溶藻弧菌Vibrio alginolyticus)胞外产物对TOCC的毒性分析。【结果】稳定传代的TOCC为成纤维样细胞,18SrDNA基因分析结果显示其来源于卵形鲳鲹;染色体核型分析发现TOCC具有二倍体特征,特征染色体数目为62条;不同培养条件下TOCC细胞的生长速率不同,28℃生长速度最快;病原菌胞外产物对TOCC具有明显的毒性。【结论】本研究成功建立广西卵形鲳鲹小脑组织来源的细胞系(TOCC)。细菌胞外产物对TOCC细胞毒性结果表明,该细胞系在一定范围内能够替代活体检测,实现体外细胞水平对广西海水养殖中病害侵染和环境污染物的快速检测,为养殖病害的有效预防提供技术支持。
