CN109662259B - 一种富益生菌冻干毛豆粒产品及其制备方法 - Google Patents
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Abstract
本发明公开了一种富益生菌冻干毛豆粒产品及其制备方法，属农产品加工技术领域。本发明以毛豆为原料，采用生成生物膜的方法将益生菌富集到毛豆表面，然后采用真空冷冻干燥的方法，制备冻干毛豆粒产品。通过此专利，可以将毛豆上富集的益生菌数从原有真空浸渍技术的106CFU/g干样提升到1010CFU/g干样，解决了毛豆上真空浸渍富集益生菌含菌量低的问题。包括以下步骤：(1)制备浸渍液：将益生菌用MRS培养到菌液含菌量在107‑9CFU/mL时，离心，取菌体，用市售纯果汁稀释菌体到含菌量为107‑9CFU/mL，将毛豆粒经清洗烫漂后；浸泡在上述浸渍液中(2)37℃静置浸泡培养4‑5天。(3)真空冷冻干燥后充氮包装，其中益生菌活菌数可在25℃下保持107CFU/g 90天以上。
Description
一、技术领域
本发明涉及一种富益生菌冻干毛豆粒产品及其制备方法，属于农产品加工技术领域。
二、背景技术
益生菌具有调节肠道菌群、降脂保肝、抗肿瘤、抗糖尿病等多种益生功能，已实现工业化应用，但其利用产品多为发酵乳制品，存在高胆固醇风险和乳糖不耐症人群无法消费两大缺陷，亟待推出新产品，惠及更多消费者。
毛豆具有降血糖、降血脂等多种保健功能，如果能将益生菌富集到毛豆中，形成富益生菌毛豆产品，同时其中的益生菌活菌数能够保持106CFU/g干样以上，此产品将兼具毛豆和益生菌的优良品质，集营养保健于一体，具有广阔市场前景。
本专利前期研究表明，采用真空浸渍制备的富益生菌毛豆其中菌数只能达到107CFU/g鲜样，经真空冷冻干燥后制成富益生菌冻干毛豆粒产品后，其中的活菌数≤106CFU/g干样。亟待找到一种新的富集方法，将毛豆中的菌数富集达到106CFU/g干样以上。
在自然状态下，微生物为抵御严苛的生存环境，具备多种自我保护机制，其中之一就是群体聚集形成生物膜。相对于浮游状态，形成生物膜的细菌抗逆性更强而且生物膜中的菌密度高，菌数多。目前的研究主要集中在致病菌的生物膜防治方面，而有益菌生物膜应用研究较少。本专利应用上述思路，提供在毛豆粒表面形成生物膜的方法来富集益生菌到毛豆上，采用此方法富集益生菌后经冷冻干燥制备的富益生菌冻干毛豆粒产品表面益生菌活菌数从原有真空浸渍技术的106CFU/g干样提升到1010CFU/g干样，解决了毛豆上真空浸渍富集益生菌含菌量低的问题。
三、发明内容
技术问题
本发明的目的是提供一种富益生菌冻干毛豆粒产品及其制备方法。
技术方案
一种富益生菌冻干毛豆粒产品及其制备方法，其制作步骤为：
(1)原料清洗烫漂：选用毛豆为原料，经清洗后，热烫30-200s后立即放入无菌冷水中，待冷却后取出沥干；
(2)益生菌浸渍液制备：取一环冻干或斜面保存的益生菌，主要为植物乳杆菌、鼠李糖乳杆菌等，接到对应的液体培养基中，在37℃培养箱中活化24h，移出1mL于新的5mL液体培养基中，同样条件活化12h，再以10％接到液体培养基中培养12h，离心后，去除上清液，用同体积市售果汁稀释，稀释菌体到含菌量为10的7-8次方，得浸渍液。
(3)形成生物膜：将烫漂后的毛豆，浸泡在上述浸渍液中，37℃静置浸泡培养4-5天。
(4)真空冷冻干燥：将已形成生物膜的毛豆经真空冷冻干燥至水分含量在5％以下。
(5)贮藏：无菌条件下，充氮铝箔袋包装，可在25℃下保持益生菌活菌数107CFU/g90天以上。
有益效果
1、与同样制备富益生菌果蔬干燥产品的专利相比，本专利的优越之处：《一种富益生菌苹果脆产品及其制备方法》(申请号：201610407046.4)该专利采用真空浸渍的方式将益生菌富集到苹果组织中，与本专利采用的富集方式完全不同。本专利曾经尝试采用真空浸渍的方式富集植物乳杆菌到毛豆中，再经真空冷冻干燥后制备富益生菌冻干毛豆粒产品，但其活菌数低于106CFU/g干样，此菌数无法达到含益生菌产品菌数要求。通过扫描电镜观察发现，毛豆子叶表面为非常光滑致密且封闭的结构，所以采用真空浸渍的方式无法将益生菌吸附到细胞间隙中。而苹果则不同，在真空浸渍时，苹果就是切成丁的，表面全部是开放的孔隙结构，非常适合益生菌富集，所以富集的菌数较高。经采用生物膜法富集益生菌后，扫描电镜显示，益生菌在毛豆的内表皮聚集形成生物膜，所以富益生菌冻干毛豆粒的含菌数提升到1010CFU/g干样，而且可在25℃下保持益生菌活菌数107CFU/g 90天以上。
2、与常规豆类脆粒产品相比较，本发明创造了一种新型的富含益生菌冻干毛豆粒产品，能够保证其中的益生菌活菌在25℃下保持107CFU/g 90天以上，本产品在保持毛豆冻干产品豆香浓郁、口感酥脆的同时，具有益生菌的调节肠道菌群、降脂保肝、抗肿瘤、抗糖尿病等多种保健功能，集保健休闲为一体，具有广阔前景。
3、与湿态的发酵豆类产品相比，本发明产品为干燥的固体形态，感官评价得分达到满意的标准，具有口感酥脆、体积小易携带、食用方便，益生菌活菌数高，可常温贮藏等优点，是一种新型的具有保健功能的休闲食品。
4、与抽真空包装和非充氮包装相比，本专利采用充氮包装可以更好保护富益生菌冻干毛豆粒中益生菌在贮藏过程中的活性，由于益生菌在贮藏中活性降低的主要主要原因为氧化损伤，所以采用充氮包装可以有效隔绝氧气，降低益生菌的氧化速度和氧化程度，从而发挥保护益生菌功能。同时充氮包装可以保护毛豆粒的外形，而真空包装容易发生碎裂的情况。
四、附图说明
1、采用真空浸渍的方式只有少量的菌富集到毛豆组织中：
下图1为真空浸渍后经真空冷冻干燥制备的毛豆粒产品的扫描电镜照片。如图可知，只能看到零星的益生菌富集到毛豆组织中，菌数测定结果表明，其中的菌数少于106CFU/g干样。
2、采用生物膜的方式可以看到大量菌体以生物膜的形态富集毛豆组织中：
下图2为形成生物膜后经真空冷冻干燥制备的冻干毛豆粒产品的扫描电镜照片。下图3为选取图2的中间部位放大的扫描电镜照片。从图可知，大量的益生菌菌体聚集成膜状，几乎填满了毛豆原有的孔隙结构，菌数测定结果表明其中的菌数达到1010CFU/g干样。
五、具体实施方式
实施例1
(1)原料清洗烫漂：选用毛豆为原料，经清洗后，热烫30-200s后立即放入无菌冷水中，待冷却后取出沥干；(2)益生菌浸渍液制备：取一环冻干或斜面保存的植物乳杆菌(Lactobacillus plantarum SICC 1.376)，接到对应的液体培养基中，在37℃培养箱中活化24h，移出1mL于新的5mL液体培养基中，同样条件活化12h，再以10％接到液体培养基中培养12h，离心后，去除上清液，用同体积市售苹果汁稀释，稀释菌体到含菌量为10的7-8次方，得浸渍液。(3)形成生物膜：将烫漂后的毛豆，浸泡在上述浸渍液中，37℃静置浸泡培养4-5天。(4)真空冷冻干燥：将已形成生物膜的毛豆经真空冷冻干燥至水分含量在5％以下，此时富益生菌冻干毛豆粒中植物乳杆菌活菌数为6.25×1010CFU/g干样。(5)贮藏：无菌条件下，充氮铝箔袋包装，可在25℃下保持益生菌活菌数107CFU/g 90天以上。
实施例2
(1)原料清洗烫漂：选用毛豆为原料，经清洗后，热烫30-200s后立即放入无菌冷水中，待冷却后取出沥干；(2)益生菌浸渍液制备：取一环冻干或斜面保存的植物乳杆菌(Lactobacillus plantarum SICC 1.376)，接到对应的液体培养基中，在37℃培养箱中活化24h，移出1mL于新的5mL液体培养基中，同样条件活化12h，再以10％接到液体培养基中培养12h，离心后，去除上清液，用同体积市售桃汁稀释，稀释菌体到含菌量为10的7-8次方，得浸渍液。(3)形成生物膜：将烫漂后的毛豆，浸泡在上述浸渍液中，37℃静置浸泡培养4-5天。(4)真空冷冻干燥：将已形成生物膜的毛豆经真空冷冻干燥至水分含量在5％以下，此时富益生菌冻干毛豆粒中植物乳杆菌活菌数为1.3×1010CFU/g干样。(5)贮藏：无菌条件下，充氮铝箔袋包装，可在25℃下保持益生菌活菌数107CFU/g 90天以上。
实施例3
(1)原料清洗烫漂：选用毛豆为原料，经清洗后，热烫30-200s后立即放入无菌冷水中，待冷却后取出沥干；(2)益生菌浸渍液制备：取一环冻干或斜面保存的鼠李糖乳杆菌(L.rhamnosus GG)，接到对应的液体培养基中，在37℃培养箱中活化24h，移出1mL于新的5mL液体培养基中，同样条件活化12h，再以10％接到液体培养基中培养12h，离心后，去除上清液，用同体积市售苹果汁稀释，稀释菌体到含菌量为10的7-8次方，得浸渍液。(3)形成生物膜：将烫漂后的毛豆，浸泡在上述浸渍液中，37℃静置浸泡培养4-5天。(4)真空冷冻干燥：将已形成生物膜的毛豆经真空冷冻干燥至水分含量在5％以下，此时富益生菌冻干毛豆粒中植物乳杆菌活菌数为4.65×1010CFU/g干样。(5)贮藏：无菌条件下，充氮铝箔袋包装，可在25℃下保持保持益生菌活菌数107CFU/g 90天以上。
实施例4
(1)原料清洗烫漂：选用毛豆为原料，经清洗后，热烫30-200s后立即放入无菌冷水中，待冷却后取出沥干；(2)益生菌浸渍液制备：取一环冻干或斜面保存的鼠李糖乳杆菌(L.rhamnosus GG)，接到对应的液体培养基中，在37℃培养箱中活化24h，移出1mL于新的5mL液体培养基中，同样条件活化12h，再以10％接到液体培养基中培养12h，离心后，去除上清液，用同体积市售桃汁稀释，稀释菌体到含菌量为10的7-8次方，得浸渍液。(3)形成生物膜：将烫漂后的毛豆，浸泡在上述浸渍液中，37℃静置浸泡培养4-5天。(4)真空冷冻干燥：将已形成生物膜的毛豆经真空冷冻干燥至水分含量在5％以下，此时富益生菌冻干毛豆粒中植物乳杆菌活菌数为5.84×1010CFU/g干样。(5)贮藏：无菌条件下，充氮铝箔袋包装，可在25℃下保持保持益生菌活菌数107CFU/g 90天以上。
以上详细说明了本发明的实施方式，但这只是为了便于理解而举的实例，不应被视为是对本发明范围的限制。同样，任何所属技术领域的技术人员均可根据本发明的技术方案及其较佳实施例的描述，做出各种可能的等同改变或替换，但所有这些改变或替换都应属于本发明的权利要求的保护范围。
Claims (2)
1.一种富益生菌冻干毛豆粒制备方法，其特征在于，是由以下方法制备而成的：
以毛豆为原料，经清洗烫漂后，以形成生物膜的方式将益生菌富集到毛豆组织中，采用真空冻干干燥方法将富益生菌毛豆粒干燥为富益生菌冻干毛豆粒，无菌条件下，充氮铝箔袋包装为产品；所述形成生物膜的方式为：(1)制备浸渍液，取一环冻干或斜面保存的益生菌，所采用的益生菌为植物乳杆菌、鼠李糖乳杆菌，接到MRS液体培养基中，在37℃培养箱中活化24h，移出1mL于新的5mL液体培养基中，同样条件活化12h，再以10％接到液体培养基中培养12h，菌液含菌量在107-9CFU/mL，离心后，去除上清液，用市售果汁稀释菌体到含菌量为107-9CFU/mL，将毛豆粒经清洗烫漂后，浸泡在上述浸渍液中；(2)37℃静置浸泡培养4-5天；所述干燥及储藏方式为真空冷冻干燥后充氮包装，其中益生菌活菌数可在25℃下保持107CFU/g 90天以上。
2.通过权利要求1所述方法制备的富益生菌冻干毛豆粒。
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