中国林蛙皮中胶原蛋白的提取研究

采用酶解法从中国林蛙皮中提取胶原蛋白。考查了提取次数、液料比、酶解时间、酶解温度以及酶用量对胶原蛋白提取率的影响。单因素考察结合正交试验设计得到胶原蛋白的优化提取条件为:提取次数为2次,液料比为60,提取时间为72 h,提取温度为37℃,酶用量为1.5%。此条件下中国林蛙皮胶原蛋白的产率可达26.2%。     

Fatty acid composition, extraction, fractionation, and stabilization of bullfrog (Rana catesbeiana) oil

The oil extracted from the fat-storage organ (fat body) of the bullfrog (Rana catesbeiana) was characterized for its fatty acid composition. The main fatty acids were palmitic (18.1%), stearic (4.1%), myristic (2.7%), oleic (31.7%), and linoleic (12.9%) acids. Long-chain polyunsaturated fatty acids were also present in significant amounts, i.e., eicosapentaenoic (1.5%) and docosahexaenoic (4.7%), and were probably derived from the fish meal content of the diet. A partially fractionated oil was extracted from the homogenized and frozen fat body with an oleic acid content of 43.2%. The natural alkaloid boldine, added at 0.5 mg/g oil level, improved the oxidative stability by a factor ranging from 1.7 to 2.4, as assessed by the Oil Stability Index method between 90 and 110°C. The stabilization effect of boldine was higher than that of naringenin, morin, and quercitin and for the synthetic antioxidant butylated hydroxytoluene at the same concentration level.     

微波辅助提取林蛙卵油的工艺优化及脂肪酸组成分析

以林蛙卵为原料,以无水乙醇为提取剂,研究微波法对林蛙卵油提取率的影响。在单因素试验的基础上,进行响应面优化试验,研究料液比、浸泡时间、微波功率对林蛙卵油提取的影响。结果表明,提取最佳工艺条件为料液比1∶9.82(g/mL)、浸泡时间3.33 h、微波时间30 min、微波功率600 W;林蛙卵油的提取率可达到(20±0.67)%。与其他林蛙卵油提取方法比较,微波法辅助提取林蛙卵油具有得率高的优点。对得到的林蛙卵油采用气相色谱-质谱分析检测出27种脂肪酸,不饱和脂肪酸质量分数54.98%,其中以亚油酸、二十碳二烯酸、二十碳三烯酸、二十碳四烯酸、亚麻酸、棕榈反油酸、二十二碳五烯酸含量较高。     

林蛙脑蛋白酶解物抗氧化性研究

用木瓜蛋白酶对林蛙脑进行酶解,并以抗坏血酸为对照,测定林蛙脑蛋白酶解液清除DPPH.、羟自由基和过氧化氢的能力.结果表明:林蛙脑蛋白酶水解液对DPPH.、.OH和H2O2均具有清除作用,尤其对H2O2有较强的清除作用,且均随浓度增加而增强.林蛙脑蛋白酶解液在浓度达到25 mg/mL时,对DPPH.的清除率达67.8%;在浓度达到4 mg/mL时,对H2O2的清除率达91.62%;浓度在4 mg/mL时,对.OH的清除率为66.89%.这说明林蛙脑蛋白酶解液具有很好的抗氧化性.     

林蛙卵雌二醇的超临界CO_2萃取及酶联免疫分析

以长白山林蛙卵为原料,研究超临界CO2萃取林蛙卵中的雌二醇的工艺条件。探讨萃取压力、萃取温度、CO2流量和萃取时间对雌二醇萃取率的影响,并采用酶联免疫分析法检测林蛙卵雌二醇的含量。结果表明最佳萃取条件为萃取压力30 MPa,萃取温度47℃,CO2流量为12 kg/h,萃取时间为90 min。酶联免疫分析得到林蛙卵中雌二醇的含量为38.11 ng/L。     

抗氧化活性林蛙胎卵多肽酶解酶的筛选

以东北林蛙胎卵为原料,以酶解液对DPPH自由基和羟自由基(.OH)清除率为指标,分别研究木瓜蛋白酶、碱性蛋白酶、中性蛋白酶、复合蛋白酶和胰蛋白酶在最佳酶作用条件下酶解时间对酶解液清除DPPH自由基、.OH的影响,筛选出适合生产抗氧化活性林蛙胎卵蛋白肽的碱性蛋白酶、木瓜蛋白酶、中性蛋白酶,再经过复合双酶酶解林蛙胎卵实验,结果表明,以中性蛋白酶与碱性蛋白酶复合酶解林蛙胎卵3h得到肽液的.OH清除率与DPPH自由基清除率最高,达到80.91%和70.45%。     

林蛙卵油的提取、成分分析及抗氧化活性研究

以林蛙卵为原料,采用超临界CO₂提取法提取林蛙卵中的林蛙卵油,研究其成分含量和抗氧化活性。采用气相色谱-质谱法(GC-MS)联用对林蛙卵油的成分进行检测分析,确定成分及含量,并进行DPPH自由基清除能力试验、羟基自由基清除试验和超氧阴离子自由基清除试验确定林蛙卵油的抗氧化能力。通过单因素实验和正交试验确定最佳提取工艺为提取压力35 MPa,提取温度60 ℃,CO₂流量10 L/h,提取时间150 min,此时林蛙卵油的提取率为34.26%。GC-MS结果显示林蛙卵油中含有34种脂肪酸,其中饱和脂肪酸和不饱和脂肪酸相对含量分别为31.95%和68.05%。体外抗氧化试验结果表明林蛙卵油对DPPH自由基的IC₅₀值为0.897 mg/mL,对羟基自由基的IC₅₀值为1.392 mg/mL,对超氧阴离子的IC₅₀值为1.687 mg/mL。林蛙卵油具有较强的体外抗氧化活性,在天然抗氧化剂和保健食品中有一定的开发利用价值。     

牛蛙皮肤抗菌肽的分离纯化与活性测定

目的分离纯化牛蛙皮肤抗菌肽,并检测其活性。方法通过刺激与浸提的方法,得到牛蛙皮肤抗菌肽粗提物,测定其抑菌活性及最低抑菌浓度。利用SephadexG-50凝胶过滤和反相高效液相色谱(RP-HPLC)分离纯化粗提物,用质谱仪测定多肽的相对分子质量及氨基酸序列,并进行活性检测。结果两种方法提取的牛蛙皮肤抗菌肽粗提物均具有较强的抑菌活性,纯化后的多肽相对分子质量约为4370,N-端含22个氨基酸,与其他两栖类来源的抗菌肽同源性较低。活性试验表明均无胰蛋白酶水解活性,存在一定的胰蛋白酶抑制剂活性,有轻微的溶血活性,不具有局部出血活性,具有抗凝和溶栓作用。结论已成功分离并纯化了1种新的牛蛙皮肤抗菌肽。     

用激光微束摘除泽蛙卵的雌核染色体产生雄核单倍体

实验表明:用红宝石激光微束摘除泽蛙卵的雌核染色体产生雄核单倍体具有准确性高、对卵子损伤小、获得单倍体胚胎多等优点。     

Effect of predator diet on life history shifts of red-legged frogs,Rana aurora

Larval red-legged frogs (Rana aurora) are known to exhibit antipredator behavior in response to both chemical alarm cues released from injured conspecifics and chemical cues of predators. In some cases, the response to predators is dependent on the predator's diet. In this experiment, we tested whether long-term exposure to predator chemical cues and alarm cues resulted in alteration of life history characteristics of red-legged frogs. We raised groups of tadpoles in the presence of chemical cues of predators that were either fed conspecifics or heterospecific caddis-fly larvae, chemical cues of injured conspecifics, or a no-cue control. Tadpoles raised in the presence of either a predator fed conspecifics or cues of injured conspecifics metamorphosed earlier and at a smaller size than those exposed to predators fed heterospecifics or the no-cue control. The result suggests that red-legged frogs exhibit a life history shift in response to predatory cues and that this response is dependent on the diet of predators.