皱纹盘鲍HdhTPX2基因在毕赤酵母中的表达及抗氧化活性研究

为研究皱纹盘鲍Haliotis discus hannai Ino硫氧还蛋白过氧化物酶（TPX2）蛋白的抗氧化活性,将HdhTPX2基因克隆至pPIC9K载体,通过电击转化至毕赤酵母GS115菌株中,获得重组表达质粒pPIC9K-HdhTPX2,经甲醇诱导及亲和层析纯化得到重组HdhTPX2蛋白,并进行质谱鉴定及体外抗氧化功能检测。结果表明:本研究中成功构建了重组毕赤酵母菌株GS115/pPIC9K-HdhTPX2,经表达条件的优化,在pH为7的培养基中用0.5%甲醇诱导表达72 h,分泌表达上清液中得到相对分子质量约为25 000的稳定表达产物,纯化后的蛋白经质谱鉴定为目的蛋白;体外活性测定发现,该蛋白清除羟自由基（·OH）能力强于维生素C,并对H2O2引起的细胞损伤具有一定的保护作用。研究表明,皱纹盘鲍硫氧还蛋白过氧化物酶HdhTPX2在毕赤酵母表达系统中得到高效表达,重组表达产物具有抗氧化活性功能。福建省自然科学基金资助项目(2017J01041);;\n国家海洋公益性行业科研专项(201405016);;\n福建省海洋生物增养殖与高值化利用重点实验室开放课题(2015fjscq04

鱼油的绿色制备工艺、标准规范 及营养功效研究进展Progress on green preparation process, standard specifications and nutritional efficacy of fish oil

鱼油富含ω-3多不饱和脂肪酸、维生素A、维生素D和天然色素等营养物质，具有多种营养功效。为促进我国鱼油产业的绿色可持续发展，保障产品质量安全，结合国内外研究进展，分别从鱼油的绿色制备工艺、标准规范和营养功效等方面进行综述，梳理了应用于鱼油生产的水酶法、超临界流体萃取法、超声辅助提取法、微波辅助提取法等绿色制备工艺，基于国内外鱼油相关的标准规范，阐述了鱼油的基本成分和质量指标、污染物限量和添加剂限量等，探讨了鱼油在预防心血管疾病、抗炎和预防脑部疾病方面的营养功效，并针对鱼油应用中存在的问题进行了分析和展望。未来应深入研究鱼油制备新工艺，完善相关标准规范，并进一步探究其生物活性机制。 Fish oil is rich in nutrients such as omega-3 polyunsaturated fatty acids (ω-3 PUFA), vitamin A/D and natural pigments, and has a variety of nutritional efficacy. To promote the green and sustainable development of the fish oil industry of China and ensure the safety and quality of fish oil products, the green preparation process, standard specifications and nutritional efficacy of fish oil were reviewed based on recent progresses at home and abroad. Several green preparation processes, including aqueous enzymatic extraction, supercritical fluid extraction, ultrasound-assisted extraction, microwave-assisted extraction methods were analyzed. Based on the relevant standard specifications of fish oil at home and abroad, the basic components and quality indicators, pollutant limits, and additive limits of fish oil were elaborated. The nutritional efficacy of fish oil in preventing cardiovascular diseases, anti-inflammation and preventing brain diseases were investigated, and the issues present in the application of fish oil were analyzed and prospected. In the future, it is necessary to conduct extensive research on the development of new preparation process for fish oil, as well as to improve the related standard specifications. Furthermore, it is essential to explore the biological activity mechanisms of fish oil in greater depth

水酶法提取紫海胆性腺脂质的工艺优化 及提取方法对其品质的影响Process optimization of lipid extraction from the gonad of Heliocidaris crassispina via enzyme-assisted aqueous method and effect of extraction methods on its quality

以福建紫海胆（Heliocidaris crassispina）性腺为研究对象，采用单因素实验和正交实验优化水酶法提取紫海胆性腺脂质工艺，并基于气相色谱-质谱联用（GC-MS）、二醇基硅胶固相萃取（Generik Diol SPE）、高效薄层色谱（HPTLC）和差示扫描量热（DSC）等技术，以脂质得率、理化性质、脂肪酸组成、营养指标、脂质组成、熔融与结晶特性为指标，探讨了水酶法、Folch法和Matyash法对紫海胆性腺脂质品质的影响。结果表明：水酶法提取紫海胆性腺脂质最佳工艺条件为料液比1∶ 20、中性蛋白酶添加量7 500 U/g、酶解时间5 h、酶解温度50 ℃和pH 7，该条件下脂质得率为2322%，显著低于Folch法（27.27%）和Matyash法（27.14%）；与Folch法和Matyash法相比，水酶法提取的紫海胆性腺脂质过氧化值（0.006 g/100 g）较低，碘值（95.77 g/100 g）较高，饱和脂肪酸含量（51.48%）较低，多不饱和脂肪酸含量（34.53%）较高，EPA与DHA总含量（15.04%）较高，营养指标较好，磷脂含量（4.23%）较低，熔融与结晶温度较低。综上，虽然水酶法提取紫海胆性腺脂质工艺存在得率低、破乳工艺复杂、磷脂选择性差等技术瓶颈，但其反应条件温和、绿色安全，所提取的脂质品质佳，具有较好的营养价值、较低的熔融与结晶温度，适合高品质海洋膳食脂质的研制。 The lipids from the gonad of Heliocidaris crassispina (Fujian Province, China) was extracted using enzyme-assisted aqueous extraction method (EAE), and the extraction process was optimized by single factor experiment and orthogonal experiment. By applying gas chromatography-mass spectrometry (GC-MS), Generik Diol solid-phase extraction (SPE), high-performance thin layer chromatography (HPTLC) and differential scanning calorimetry (DSC) technologies, the quality of EAE lipid was comparatively characterized with that of Folch method and Matyash method with the lipid yields, physicochemical properties, fatty acid compositions, nutritional indexes, lipid compositions, melting and crystallization characteristics as indicators. The results showed that the optimal conditions of EAE process were determined as follows: solid-liquid ratio 1∶ 20, neutral protease dosage 7 500 U/g, enzymatic hydrolysis temperature 50 ℃，enzymatic hydrolysis time 5 h, and pH 7. The lipid yield was 23.22% under above conditions, which was significantly lower than that of Folch method (2727%) and Matyash method (27.14%). Compared with Folch method and Matyash method, the lipids from the gonad of Heliocidaris crassispina extracted by EAE had lower peroxide value (0.006 g/100 g), higher iodine value (95.77 gI/100 g), lower content of saturated fatty acids (51.48%), higher content of polyunsaturated fatty acids (34.53%), higher total content of EPA and DHA (15.04%), better nutritional indexes, lower phospholipids content (4.23%) and lower melting temperature and crystallization temperature. In conclusion, though bearing the shortages of low yield, complicated demulsification process and poor selectivity of phospholipids, EAE surpassed Folch method and Matyash method for milder conditions, eco-friendly initiatives, better quality, better nutritional value, lower melting and crystallization temperature of lipids, and suitable for the development of high quality marine dietary lipids