Liang Xin College, China Jiliang University, Hangzhou, Zhejiang 310018, People's Republic of China.
College of Life Sciences, China Jiliang University, Hangzhou, Zhejiang 310018, People's Republic of China.
Int J Med Mushrooms. 2021;23(5):41-53. doi: 10.1615/IntJMedMushrooms.2021038136.
Response surface methodology with a Box-Behnken design was used to investigate the effects of a neutral protease on Auricularia auricula-judae polysaccharide (AAP) extraction yield. Four ethanol concentrations (30%, 50%, 70%, and 90%) were used to grade AAPs. Optimal fractions with a high polysaccharide recovery rate and a strong free radical scavenging rate were obtained, and their antioxidant stress effect was studied using Caenorhabditis elegans as the model organism. The results showed that among the factors affecting AAP yield (temperature, liquid-to-A. auricula-judae powder, enzyme usage, time, pH, etc.), temperature, ratio of liquid-to-A. auricula-judae powder, and time had the most obvious effects. AAP yield reached 13.91% ± 0.95% at a temperature of 7°C, liquid-to-material ratio of 75 mL/g, and extraction time of 70 min. High AAP recovery rates and strong free radical scavenging rates of 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) diammonium salt and 1,1-dibenzoyl-2-trinitrophenylhydrazine were obtained by 30% ethanol (AAPs1). Results from the anti-hydrogen peroxide and methyl viologen stress experiment showed that AAPsl could significantly prolong the life of worms under oxidative stress compared to the negative control group (P < 0.05).
响应面法中的 Box-Behnken 设计被用于研究中性蛋白酶对黑木耳多糖（AAP）提取率的影响。采用 30%、50%、70%和 90%四种乙醇浓度对 AAP 进行分级。获得了高多糖回收率和强自由基清除率的最佳级分，并以秀丽隐杆线虫为模型生物研究了它们的抗氧化应激作用。结果表明，在影响 AAP 产量的因素（温度、液料比、酶用量、时间、pH 等）中，温度、液料比和时间的影响最为显著。在温度为 7°C、液料比为 75mL/g、提取时间为 70min 的条件下，AAP 的产率达到 13.91%±0.95%。通过 30%乙醇（AAPs1）可获得高 AAP 回收率和强 2,2'-偶氮双（3-乙基苯并噻唑啉-6-磺酸）二铵盐和 1,1-二苯甲酰基-2,4,6-三硝基苯肼自由基清除率。过氧化氢和甲基紫精应激实验结果表明，与阴性对照组相比，AAPsl 能显著延长氧化应激下蠕虫的寿命（P<0.05）。
