摘要
本研究中黄伞液体发酵终点为144h。菌丝体鲜重达40.1g/100mL,发酵液还原糖含量5.0mg/mL,发酵液的终点pH为5.4。
运用不同物理方法对黄伞发酵醪液多糖进行提取,在所选择的四种方法：超声波辅助加压法、微波法、高压法、超声波法中,超声波辅助加压法所得提取液黄伞多糖含量显著高于其余三种；超声波辅助加压法的最佳参数为：超声波破壁时间35min、压力0.12MPa、提取时间为40min,在该条件下,提取液多糖浓度为9.5mg/mL。将黄伞发酵醪液提取后,以蔗糖将提取液含糖量调整到12.5%,经发酵法得到黄伞菌丝体醋。在酒精发酵阶段,发酵适宜的条件为：酵母接种量10%,适宜发酵温度30℃,发酵时间80h,发酵液的酒精含量可达7.0%。醋酸发酵阶段,发酵适宜条件为：起始pH值5.5,接种量11%,温度33℃,摇瓶装量200mL(500mL摇瓶),摇床转速160r/min,发酵时间80h；发酵液的酸度(以醋酸计)为3.52g/100mL,同时酒精含量降至0.9%。
在澄清实验中,采用明胶、皂土、壳聚糖、皂土+壳聚糖、明胶+壳聚糖对黄伞菌丝体醋进行澄清研究。结果表明：皂土+壳聚糖为最佳澄清剂,经皂土+壳聚糖处理的黄伞菌丝体醋透光率达到96%,还原糖保存率为96.5%,总酸保存率95.8%。黄伞菌丝体醋的用量为12%,白砂糖的最佳添加量为8%,蜂蜜的最佳添加量为5%,可获得酸甜爽口、风味独特的黄伞菌丝体醋饮料。黄伞菌丝体醋饮料的灭菌条件选择巴氏消毒灭菌,高温老化实验后,各项指标稳定,稳定性较高。黄伞菌丝体醋饮料中总酸(以醋酸计)含量为≥0.42g/100mL,可溶性固形物为≥10%,多糖含量为≥1.44 mg/mL,总糖浓度≥11.2%。黄伞菌丝体醋饮料色泽浅黄色,卫生指标符合国家标准。
对黄伞菌丝体醋饮料进行动物功能学检测表明：通过小鼠腹腔巨噬细胞吞噬鸡红细胞实验,表明黄伞菌丝体醋饮料具有一定的增强免疫力功效,增强免疫力效果随着饮料剂量的增加而升高,表现出一定的量效关系。
Studies on the shaking flask fermentation of Pholiota adiposa showed that the biomass amount reached over 40.1g/100mL (fresh weight) after having been fermented for 144h; the residual sugar content of the fermentation liquid was about 5.0mg/mL, and the PH of the end point was 5.4.
Four physical methods were studied to detect the optimum extraction conditions of polysaccharides from Pholiota adiposa mycelia and fermentation liquid, including the combination of ultrasonic extraction and high pressure, microwave method, high pressure method, and ultrasonic method.The results showed that the combination of ultrasonic extraction and high pressure could get more polysaccharides than the other methods. The optimum parameters of the combination of ultrasonic extraction and high pressure method was as follows:the time of ultrasonic treatment was 35min, the pressure was 0.12MPa, and extraction time was 40min. After the combination extracting, the amount of polysaccharides reached 9.5mg/mL. After the extraction, the sugar amount of the Pholiota adiposa fermented liquid was 12.5% by adding sugar. After that, they were fermented to vinegar beverage through Alcohol and Acetobacter fermentation phases. The optimum fermentation conditions were obtained when the inoculating amount of liquid seed was 10%, culture temperature of 30℃,80h respectively in Alcohol fermentation and the amount of alcohol could reach to 7.0%. The optimum fermentation conditions were obtained when the inoculating amount of liquid seed was 11%, culture temperature of 33℃, the original pH 5.5, the shaker rotating at 160r/min, the amount of culture medium 200mL(500mL triangle flask),80h respectively in Acetobacter fermentation. The amount of acetic acid could reach to 3.52g/100mL, and alcohol declines to 0.9% in the meantime.
In the clarifying experiments, clarifiers including gelatin, bentonite, chitosan, bentonite and chitosan, and gelatin and chitosan were used respectively to clarify Pholiota adiposa mycelia fermentation vinegar. The results indicated that bentonite and chitosan was the best clarifier. The relative parameters of the fermentation vinegar clarified by bentonite and chitosan were as follows:light transmittance reaches 96%, residual sugar preservation rates was 96.5%, and total acid preservation rate was 95.8%. A sour and sweet and tasty vinegar beverage was obtained by combining 8% white sugar,5% honey,and 12% Pholiota adiposa mycelia fermentation vinegar.
Pasteurization were chosen as the sterilization conditions, the result of the aging experiment showed that the index signs of the product kept changeless and its stability is higher. In the finished product of vinegar beverage from Pholiota adiposa, the amount of acid(with the acetic acid account) reached to≥0.42g/100 mL, the dissolubility solid was≥10% and polysaccharides concentration was≥1.44 mg/mL, total polysaccharide concentration was≥11.2%. The beverage, with a buff color, accords with the national hygienic standard.
The animal study showed that vinegar beverage from Pholiota adiposa was given orally to mice 30 days in order to measure the phagocytic index and phagocytic rate of the macrophage of mice. The experiment showed that the vinegar beverage could enhance the immunity. The enhancement of immunity is enhanced with the increment of the dosages.
引文
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