Isolation and Identification of Starch Flocculants in Quinoa Acid Pulp and Its Application in Quinoa Protein Extraction
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摘要: 采用平板分离技术,以淀粉絮凝率和产乳酸量为指标,从自然发酵的藜麦酸浆中分离筛选出四株产乳酸细菌。根据其形态学特征、生理生化实验以及16S rDNA基因序列分析,鉴定菌株QSL4为屎肠球菌(Enterococcus faecium)、QSL8为耐久肠球菌(Enterococcwus durans)、QSL9为蒙氏肠球菌(Enterococcus mundtii)、QSL12为乳肠球菌(Enterococcus lactis)。将分离出的菌株应用于藜麦蛋白的提取中,通过单因素实验,对其在藜麦蛋白中的提取特性进行分析,结果表明:当接种量为7%,温度为26 ℃时,菌株QSL12对藜麦蛋白的提取率最高,分别达到60.23%和62.39%,均高于碱提酸沉法52.13%。本研究结果为酸浆法提取藜麦蛋白的深入研究奠定基础。Abstract: In this study, four strains of lactic acid-producing bacteria were isolated from naturally fermented quinoa sour pulp by using plate separation technology with starch flocculation rate and lactic acid production as indicators. According to the morphological characteristics, physiological and biochemical experiments, and 16S rDNA sequence analysis, strain QSL4, QSL8, QSL9 and QSL12 were identified as Enterococcus faecium, Enterococcwus durans, Enterococcus mundtii and Enterococcus lactis, respectively. The isolated strains were applied to the extraction of quinoa protein, and its extraction characteristics in quinoa protein were analyzed by single factor experiments. The results showed that the extraction rate of quinoa protein by strain QSL12 was the highest, reaching 60.23% and 62.39% at the inoculum level of 7% and the fermentation temperature was 26 ℃, which were higher than the extraction rate results of 52.13% by alkali dissolution and acid precipitation method. This study would provide a foundation for quinoa protein extraction by acidic steeping liquor (ASL).
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Key words:
- quinoa sour pulp /
- separation and identification /
- starch /
- flocculation /
- quinoa protein
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表 1 菌株特征
Table 1. Strain characteristics
菌株 透明度 菌落形状 菌落边缘 菌落表面 颜色 隆起 直径(mm) 细胞
形态排列 革兰氏
染色QSL4 半透明 圆 整齐 光滑 乳白色 低 0.5 球形 单个
成堆G+ QSL8 不透明 圆 整齐 光滑 乳黄色 低 1.1 球形 单个
成堆G+ QSL9 不透明 圆 整齐 光滑 乳黄色 低 0.5 球形
椭圆成对
链状G+ QSL12 半透明 圆 整齐 光滑 乳白色 低 0.3 球形 成对
链状G+ 注:“G+”表示革兰氏阳性细菌。 表 2 四株菌株的生理生化实验结果
Table 2. Physiological and biochemical characterization of four strains
菌株 QSL4 QSL8 QSL9 QSL12 过氧化氢酶 ? ? ? ? 运动性实验 ? ? ? ? H2S生成实验 ? ? ? ? 硝酸盐还原实验 + + ? + 明胶液化 ? ? ? ? 葡萄糖产气 + ? + ? 果糖 + + + + 蔗糖 + + + + 乳糖 + + + + 半乳糖 + + + + 麦芽糖 + + + + 甘露醇 + + + + 山梨醇 ? + + + 甘露糖 ? + + + 纤维二糖 + + ? + L-阿拉伯糖 ? ? ? ? 葡萄糖酸盐 + + + + 吲哚实验 ? ? ? ? 精氨酸产氨 + + + + 注:“+”表示实验结果为阳性;“?”表示实验结果为阴性。 表 3 菌株的同源相似性分析
Table 3. Homology similarity analysis of strains
菌株 序列长度(bp) 同源菌株 登录号 同源性(%) QSL4 2689 Enterococcus faecium strain
GXNN20201226-1MZ452347.1 100 QSL8 2366 Enterococcus durans strain 4528 MT545044.1 98.41 QSL9 2444 Enterococcus mundtii strain
DE-5MT240916.1 98.01 QSL12 2547 Enterococcus sp. strain Y3-O-1 OK326673.1 100 -
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