CN105802876A - 一种复合益生菌发酵苜蓿嫩芽粉制剂及其制备方法和应用 - Google Patents
一种复合益生菌发酵苜蓿嫩芽粉制剂及其制备方法和应用 Download PDFInfo
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Abstract
本发明提供一种复合益生菌发酵苜蓿嫩芽粉制剂及其制备方法和应用,以植物乳杆菌SCI‑01、植物乳杆菌P‑8、动物双歧杆菌乳亚种V9和干酪乳杆菌Zhang为发酵菌剂,其保藏编号分别为CGMCC No.11930、CGMCC No.6312、CGMCC No.5470和CGMCC No.5469;发酵培养基组成:以重量份数计,苜蓿嫩芽粉970‑1000,蔗糖8‑12,大豆蛋白粉8‑12,料水比1:1;初始pH 6.0,总接种量为1×107CFU/g,37℃发酵,pH 5.0为发酵终点;发酵产物经真空冷冻干燥获得苜蓿益生菌原粉制剂,益生菌活菌数达到1.0×1010CFU/g以上,直接或与木糖醇调配后可制成片剂、粉剂或胶囊剂。
Description
技术领域
本发明涉及一种复合益生菌发酵制剂,尤其涉及一种由植物乳杆菌SCI-01、干酪乳杆菌Zhang、乳双歧杆菌V9和植物乳杆菌P-8混和发酵的苜蓿嫩芽粉制剂及其制备方法和应用,属于益生菌发酵制剂技术领域。
背景技术
益生菌是一类通过改善肠道微生物从而对宿主产生有益影响的微生物。它可通过参与肠道微生物平衡来抑制病原菌生长,避免了长期使用抗生素产生的耐药性和二重感染,因此,益生菌产品近年来备受人们的关注。目前,虽然市场上的益生菌产品极其丰富,但是开发的产品以发酵乳制品为主,其中含有高的脂肪和胆固醇,还可能引发乳糖不耐症。所以,非乳益生菌产品成为发展的趋势。而豆科植物苜蓿,具有很高的营养和保健价值,作为一种天然碱性食物,含有大量的苜蓿叶蛋白,膳食纤维,活性多糖,皂甙,黄酮,维生素E,硒,叶黄素等天然活性成分,其营养保健功能及其所含的生理活性已成为营养保健及提取药品的宝贵资源。所以,以苜蓿为基质的发酵食品将越来越受到人们的关注。
苜蓿草嫩芽的叶蛋白含有18种氨基酸,包括8种人体必需氨基酸,且组成比例均衡,与***粮农组织推荐的成人氨基酸模式基本相符,特别是赖氨酸,叶黄素含量很高,并且富含大量的胡萝卜素、必需脂肪酸、多种维生素,以及硒,铁、镁、钙、锌、铜等矿物质元素,具有防病治病、防衰抗老、强身键体,显著提高人体内血红蛋白的含量等多种生理功能。
苜蓿草嫩芽的膳食纤维主要包括纤维素、半纤维素、果胶及果胶类物质和木质素四个部分。膳食纤维具有重要的生理功效,包括:膳食纤维能结合肠腔中的诱癌物质,减少粪便停留时间,进而减少大肠粘膜与诱癌物质的接触,使粪便***并增加粪便的排出量,可预防便秘、结肠癌、肠憩室、痔疮和下肢静脉曲张;降低血清胆固醇,预防由冠动脉硬化引起的心脏病;改善末梢神经组织对胰岛素的感受性,能调节糖尿病人的血糖水平;是一种无能量填充料,能防治肥胖症;膳食纤维的缺乏或不足还与阑尾炎、胆结石、膀胱结石、十二指肠溃疡、溃疡性结肠炎和乳腺癌等疾病的发病率与发病程度有很大的关系。
苜蓿草嫩芽的植物多糖尤其是经异构化或修饰后,具有很好的抗肿瘤作用,对T细胞、B细胞活化的免疫有增强作用,还可以降血脂、降血糖、解毒、抗菌、抗辐射。苜蓿多糖能够提高机体淋巴细胞产生抗体的能力。
苜蓿草嫩芽的黄酮类化合物是中草药的主要有效成分之一,(1)儿茶素能促进动物的生长,增加细胞中酶的功能,(2)抗氧化和清除自由基的作用,(3)对重金属有螯合作用,因此在重金属污染环境中生活工作的人服用黄酮类化合物可防止重金属中毒;(4)生物黄酮有对抗肿瘤细胞疯长的作用,而且对致癌因子和促癌因子有生化抑制作用。能防止肿瘤的发生和抑制肿瘤生长;(5)苜蓿中的黄酮是使维生素C稳定和不容易被破坏的最重要的物质。黄酮还能加强维生素C的抗氧化和清除自由基的作用。(6)生物黄酮是微生物抑制剂,对葡萄球菌、大肠杆菌、痢疾杆菌和伤寒杆菌有抑制作用,对病毒(如流感病毒、肠炎病毒)有很强的抗感染和抗炎作用。
苜蓿草嫩芽的维生素E在医药、化妆品上都有广泛的应用。维生素E是一种抗氧化剂,可以对抗氧自由基的破坏作用,抑制有毒脂质过氧化物的形成,保护细胞膜不破坏,使不饱和脂肪酸稳定,维持组织正常的新陈代谢,因此临床上有着广泛的应用。如增加机体免疫功能、延缓人体衰老、预防和治疗心血管疾病、治疗动脉硬化症和防止阿霉素引起的脱发、预防脑溢血和糖尿病等。维生素E可以进入细胞,并且具有抗自由基链式反应的作用,所以在皮肤保养上的应用上也大有前途。
苜蓿草嫩芽的皂甙类具有多种多样的药理活性和生物活性,如可抑制胆固醇在肠道的吸收,降低血浆胆固醇,防治心脑血管疾病、降压、抗菌、消炎、减热、镇静、强壮等方面的药理作用。苜蓿皂甙毒性低、药源多,开发成有用的降血脂及治疗冠心病的药物,是很有价值的。
本发明利用分别分离自泡菜、酸牛乳、健康儿童肠道和酸马奶样品中的L.plantarumSCI-01、L.plantarum P-8、B.lactis V9和L.casei Zhang益生菌株发酵苜蓿嫩芽粉。现有技术中紫花苜蓿多发酵应用于青贮饲料,尚未有此类复合益生菌发酵苜蓿嫩芽粉制剂。
专利申请CN102813004A公开了以干酪乳杆菌和双歧杆菌为发酵剂制备的豆乳饮料,其特征是配料组成为:大豆乳或大豆粉还原乳300kg、葡萄糖10~15kg、蔗糖100~120kg稳定剂2~4kg、酸液0.1~0.3kg、食用香精或香料0.1~0.3kg、干酪乳杆菌(Lactobacillus casei Zhang)发酵剂0.015~0.03kg、双歧杆菌(Bifidobacterium animalis subsp.lactis V9)发酵剂0.015~0.03kg,所述发酵剂菌株保藏于中国微生物菌种保藏管理委员会普通微生物菌种保藏中心,保藏号分别为CGMCCNo.5469和CGMCC No.5470。
专利申请CN102994422A公开了一种植物乳杆菌,其特征在于:所述植物乳杆菌(Lactobacillus plantarum P-8)菌株分离自酸牛奶中,具有优异的抗胃肠道消化液耐受能力。该菌株已保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏号:CGMCC No.6312。
发明内容
本发明目的是提供一种复合益生菌发酵苜蓿粉制剂,利用分别分离自泡菜、酸牛乳、健康儿童肠道和酸马奶样品中的L.plantarum SCI-01、L.plantarum P-8、B.lactis V9和L.caseiZhang益生菌株发酵苜蓿嫩芽粉,上述益生菌具有优异的抗胃肠道消化液耐受能力;改善人体肠道菌群结构的能力;调节血脂代谢,对高血脂症肝脏具有保护修复作用,增强机体抗氧化能力;对机体免疫具有调节作用;有效预防缓解腹泻、便秘、腹痛、腹胀等肠易激综合症;有效预防肿瘤生长和Ⅱ型糖尿病的发生等功效。利用上述复合益生菌发酵苜蓿嫩芽粉,不但能够改善和提高苜蓿的营养价值和口感,还能形成益生菌加益生元的保健功效。
本发明的目的是通过如下技术方案实现的:
一种复合益生菌发酵苜蓿嫩芽粉制剂,以植物乳杆菌SCI-01(L.plantarum SCI-01)、植物乳杆菌P-8(L.plantarum P-8)、动物双歧杆菌乳亚种V9(B.lactis V9)和干酪乳杆菌Zhang(L.casei Zhang)为发酵菌剂,其保藏编号分别为CGMCC No.11930、CGMCC No.6312、CGMCC No.5470和CGMCC No.5469;发酵培养基组成:以重量份数计,苜蓿嫩芽粉970-1000,蔗糖8-12,大豆蛋白粉8-12,料水比1:1;初始pH 6.0,接种上述四种菌剂,总接种量为1×107CFU/g,37℃发酵,pH 5.0为发酵终点。
优选的,所述发酵菌剂L.plantarum SCI-01、L.plantarum P-8、B.lactis V9和L.casei Zhang的接种量比例为1:1:1:2。
通过上述所得益生菌苜蓿发酵产物中的益生菌活菌数达到1.0×1010CFU/g以上。
所述益生菌苜蓿发酵产物与无菌处理后的保护剂按照1:2的比例混合均匀后经真空冷冻干燥制备苜蓿益生菌原粉制剂,该苜蓿益生菌原粉制剂中益生菌活菌数达1.0×1010CFU/g以上。
优选的,所述保护剂组成为:海藻糖4%,乳糖4%,大豆蛋白粉2%,蒸馏水90%。)
所述的苜蓿益生菌原粉制剂直接或与木糖醇等调配后,可以制备成苜蓿益生菌片剂、粉剂、胶囊剂等常见剂型的口服制剂。这些制剂具有易于携带和使用、益生菌含量高、易于消化吸收好、口感好的特点。经调配后的制剂中益生菌活菌数不低于2.0×109CFU/g。
本发明提供一种植物乳杆菌L.plantarum SCI-01,于2015年12月24日保藏于中国微生物菌种保藏管理委员会普通微生物中心(简称CGMCC地址:北京市朝阳区北辰西路1号院3号,中国科学院微生物研究所,邮编:100101),保藏编号:CGMCC No.11930,分类命名:植物乳杆菌Lactobacillus plantarum。
本发明提供的植物乳杆菌L.plantarum SCI-01是自300多株乳酸菌中筛选出可以抑制肠道致病细菌(大肠杆菌O157:H7、鼠伤寒沙门氏菌、弗氏志贺氏菌、金黄色葡萄球菌、单细胞李斯特氏菌)和腐败真菌(黄曲霉、寄生曲霉、娄地青霉)的优良乳酸菌株,该菌株可高产广谱抑菌物质—苯乳酸(70mg/L)。通过复配发酵发现其抑菌活性增强且对蛋白酶具有良好的稳定性以及菌株具有良好的耐受胃肠液及胆盐的特点。菌株L.plantarum SCI-01具有潜在益生特性,可作为酸奶中的生物保鲜菌开发应用。
有益效果:
四菌联用(L.casei Zhang、B.animalis V9、L.plantarum P-8、L.plantarum SCI-01),其中L.casei Zhang、B.animalis V9和L.plantarum P-8符合中国人自己的肠道特点;具有降血脂、预防腹泻、便秘;预防糖尿病;改善肠道微生态环境的益生特性。所制备的苜蓿益生菌制剂具有益生菌含量高(活性益生菌2000亿/100g),货架期内活菌活力高的特点;苜蓿嫩芽粉经4株益生菌发酵后,其有机酸、维生素、游离氨基酸、生物活性肽增多,活性异黄酮含量增加,易于人体消化吸收和健康功效的提升。
复合益生菌发酵苜蓿嫩芽粉中维生素B1和B6含量都有不同程度的提高。发酵后VB1含量为5.69ug/g,较发酵前提高65.4%;VB6含量增加显著,发酵后含量为24.59ug/g。乳酸菌发酵苜蓿嫩芽粉有利于维生素B6和B1的积累。复合益生菌发酵苜蓿嫩芽粉,能够影响苜蓿中原有的异黄酮含量。发酵后活性较弱的葡萄糖苷型异黄酮(染料木苷)含量降低30.7%,为3.88ug/g,而黄豆苷元的含量在发酵后增加,为0.44ug/g,含量较发酵前提高了76.0%。复合益生菌发酵苜蓿有利于活性异黄酮含量(黄豆苷元)的增加,更利于人体对异黄酮的吸收。
附图说明
图1:不同灭菌方式对苜蓿嫩芽粉杂菌的影响;
图2:大豆蛋白能够促进复合益生菌对苜蓿嫩芽粉的发酵;
图3:添加蔗糖能够显著提发窘苜蓿嫩芽粉益生菌含量;
图4:不同接种比例对发酵苜蓿粉益生菌含量的影响;
图5:复合益生菌发酵能够显著增加苜蓿B族维生素的含量;
图6:复合益生菌发酵能够显著增加苜蓿中活性异黄酮的含量;其中,6(a)为染料木苷含量变化,6(b)为黄豆苷元含量变化;
图7:保护剂能够显著提高发酵苜蓿粉冻干过程中的益生菌存活率。
具体实施方式
以下结合具体实施例进一步描述本发明,本发明的优点和特点将会随着描述而更为清楚。但这些实例仅是范例性的,并不对本发明的范围构成任何限制。本领域技术人员应该理解的是,在不偏离本发明的精神和范围下可以对本发明技术方案的细节和形式进行修改或替换,但这些修改和替换均落入本发明的保护范围内。下述实施例中涉及的方法、成分及用量,如无特殊说明,均为本领域技术人员所知常规方法。
下述实施例中使用的益生菌发酵菌剂均为活性冻干菌粉。
复合益生菌(L.plantarum SCI-01、L.plantarum P-8、B.lactis V9和L.casei Zhang)发酵苜蓿嫩芽粉制剂的制备方法。
将苜蓿嫩芽粉与大豆蛋白粉、蔗糖粉和蒸馏水按比例混合均匀后灭菌处理。L.plantarumSCI-01、L.plantarum P-8、B.lactis V9和L.casei Zhang发酵菌剂接种于灭菌完毕的发酵培养基中,初始pH 6.0,37℃恒温培养,pH5.0为发酵终点。
活菌计数:
一般采用倾注平板计数法,操作步骤为:依据国标方法,摇匀发酵液,吸取1mL发酵液置于9mL的无菌生理盐水中,进行10倍稀释,随后依次稀释至所需梯度。将最后稀释梯度试管稀释振匀后,吸取1mL稀释液倾注至平板中,到入培养基,摇匀,置凝固,于37℃培养箱培养48h。平板分别做两个梯度,每个梯度两个平行。
测定乳酸菌活菌数培养基:MRS固体培养基,厌氧培养。
pH测定:使用精确pH计直接测定发酵液pH值。
实施例1:
研究不同灭菌方式对苜蓿发酵培养基的影响。
固态发酵不同于液态,存在传热速度慢、传热不均匀,灭菌不彻底的难点。研究不同灭菌方式对苜蓿嫩芽粉中杂菌的杀灭效果。具体结果见图1。温度的提高,苜蓿粉中原有的杂菌逐渐降低,当灭菌条件为121℃15min时,灭菌效果最好;此外相比于细菌(主要为芽孢杆菌),真菌耐热性较低,95℃30min即可达到灭菌效果。具体见图1所示。
实施例2:
研究大豆蛋白对复合益生菌发酵苜蓿嫩芽粉的影响。
接种量和各菌剂比例相同。
a.发酵培养基:苜蓿嫩芽粉1000g,蔗糖0g,大豆蛋白粉0g,料水比1:1。发酵苜蓿粉中乳酸菌活菌数为4.93×109CFU/g。
b.发酵培养基:苜蓿嫩芽粉1000g,蔗糖0g,大豆蛋白粉10g,料水比1:1。发酵苜蓿粉中乳酸菌活菌数为7.09×109CFU/g。
试验结果表明:经过蛋白处理的苜蓿嫩芽粉,有利于乳酸菌的生长。蛋白处理后,发酵苜蓿的乳酸菌活菌数为70.9亿CFU/g,较未处理活菌数提高43.9%。具体见图2所示。
实施例3:
研究蔗糖对益生菌发酵苜蓿嫩芽粉的影响。
接种量和各菌剂比例相同。
a.苜蓿嫩芽粉1000g,蔗糖0g,大豆蛋白粉10g,料水比1:1。发酵苜蓿粉中乳酸菌活菌数为6.75×109CFU/g。
b.苜蓿嫩芽粉1000g,蔗糖10g,大豆蛋白粉10g,料水比1:1。发酵苜蓿粉中乳酸菌活菌数为1.015×1010CFU/g。
c.苜蓿嫩芽粉1000g,蔗糖20g,大豆蛋白粉10g,料水比1:1。发酵苜蓿粉中乳酸菌活菌数为1.102×1010CFU/g。
试验结果表明:添加蔗糖的苜蓿嫩芽粉,有利于乳酸菌的生长。添加后,发酵苜蓿的乳酸菌活菌数为101.5~110.2亿CFU/g,较未添加蔗糖组活菌数提高50.37~63.26%。具体见图3所示。
实施例4:
研究不同菌种接种比例对苜蓿发酵培养基的影响。
发酵培养及组成:以重量份数计,苜蓿嫩芽粉970-1000,蔗糖8-12,大豆蛋白粉8-12,料水比1:1。
a.当L.plantarum SCI-01、L.plantarum P-8、B.lactis V9和L.casei Zhang的总接种量为1×107CFU/g(接种比例1:1:1:1),37℃发酵,测定乳酸菌活菌数为0.72×1010CFU/g。
b.当L.plantarum SCI-01、L.plantarum P-8、B.lactis V9和L.casei Zhang的总接种量为1×107CFU/g(接种比例1:2:1:1),37℃发酵,测定乳酸菌活菌数为0.84×1010CFU/g。
c.L.plantarum SCI-01、L.plantarum P-8、B.lactis V9和L.casei Zhang的总接种量为1×107CFU/g(接种比例1:1:2:1),37℃发酵,测定乳酸菌活菌数为0.93×1010CFU/g。
d.L.plantarum SCI-01、L.plantarum P-8、B.lactis V9和L.casei Zhang的总接种量为1×107CFU/g(接种比例1:1:1:2),37℃发酵,测定乳酸菌活菌数为1.03×1010CFU/g。
实验结果:依据实验结果,当L.plantarum SCI-01、L.plantarum P-8、B.lactis V9和L.caseiZhang的总接种量为1×107CFU/g(接种比例1:1:1:2),更有利于苜蓿粉的发酵。具体见图4所示。
综合上述研究,复合益生菌发酵苜蓿粉的最佳发酵培养基组成为:以重量份数计,苜蓿嫩芽粉970-1000,蔗糖8-12,大豆蛋白粉8-12,料水比1:1。初始pH6.0,接种L.plantarumSCI-01、L.plantarum P-8、B.lactis V9和L.casei Zhang发酵菌剂,接种比例为1:1:1:2,总接种量为1×107CFU/g,37℃发酵,pH 5.0为发酵终点。发酵产物中益生菌活菌数达到1.0×1010CFU/g以上。此时,复合益生菌发酵苜蓿嫩芽粉中维生素B1和B6含量都有不同程度的提高。发酵后VB1含量为5.69ug/g,较发酵前提高65.4%;VB6含量增加显著,发酵后含量为24.59ug/g。乳酸菌发酵苜蓿嫩芽粉有利于维生素B6和B1的积累。复合益生菌发酵苜蓿嫩芽粉,能够影响苜蓿中原有的异黄酮含量。发酵后活性较弱的葡萄糖苷型异黄酮(染料木苷)含量降低30.7%,为3.88ug/g,而黄豆苷元的含量在发酵后增加,为0.44ug/g,含量较发酵前提高了76.0%。复合益生菌发酵苜蓿有利于活性异黄酮含量(黄豆苷元)的增加,更利于人体对异黄酮的吸收。具体结果见图5-6.
实施例5:
研究不同保护物质对冷冻干燥苜蓿益生菌制剂的影响。
a.所述的复合益生菌苜蓿发酵产物与无菌处理后的保护剂(蒸馏水)按照1:2的比例混合均匀后经真空冷冻干燥制备苜蓿益生菌原粉制剂,该制剂中益生菌活菌数达5.9×109CFU/g。
b.所述的复合益生菌苜蓿发酵产物与无菌处理后的保护剂(海藻糖4%,乳糖4%,大豆蛋白粉2%,蒸馏水90%)按照1:2的比例混合均匀后经真空冷冻干燥制备苜蓿益生菌原粉制剂,该制剂中益生菌活菌数达1.0×1010CFU/g。
实验结果:依据实验结果,当复合益生菌苜蓿发酵产物与无菌处理后的保护剂(海藻糖4%,乳糖4%,大豆蛋白粉2%)按照1:2的比例混合均匀后经真空冷冻干燥,利于苜蓿制剂中益生菌活性的保存和提高。具体见图7所示。
实施例6:
复合益生菌发酵苜蓿粉制剂的应用。
将真空冷冻干燥后的益生菌发酵苜蓿干粉经低温粉碎过100目筛后制备获得苜蓿益生菌原粉。苜蓿益生菌原粉直接或与木糖醇等调配后,可以制备成苜蓿益生菌片剂、粉剂、胶囊等常见剂型的口服制剂。
苜蓿益生菌原粉70-90份,木糖醇10-30份经压片机挤压成型为0.5-0.8g片剂。片剂中益生菌含量为不低于20亿CFU/g。
苜蓿益生菌原粉70-90份,木糖醇10-30份经粉剂包装机包装为为2.0-2.5g的粉剂。粉剂中益生菌含量为不低于70亿CFU/g。
苜蓿益生菌原粉70-90份,木糖醇10-30份经胶囊填充剂填充为0.2-0.3g的胶囊。胶囊剂中益生菌含量为不低于70亿CFU/g。
经贮藏试验发现苜蓿益生菌片剂于4℃和25℃贮藏1个月后,其益生菌存活率分别为95%和60%,具体贮藏数据见表1。苜蓿益生菌粉剂或胶囊4℃和25℃贮藏1个月后,其益生菌存活率分别为97%和75%,具体贮藏数据见表2。
苜蓿益生菌片剂、粉剂和胶囊富含蛋白质、膳食纤维,此外还含有对人体有益的黄酮类物质(木犀草素)、异黄酮物质(黄豆苷元、染料木苷),具体见表3。
表1 苜蓿益生菌片剂贮藏稳定性结果
表2 苜蓿益生菌粉剂、胶囊制剂贮藏稳定性结果
表3 苜蓿益生菌片剂营养成分
项目 | 每100克(g) |
蛋白质 | 18.43g |
脂肪 | 6.59g |
碳水化合物 | 34.42g |
钠 | 239mg |
钾 | 3.69g |
膳食纤维 | 28.70g |
维生素B6 | 0.3mg |
黄酮(木犀草素) | 0.61mg |
异黄酮(黄豆苷元) | 0.05mg |
异黄酮(染料木苷) | 0.12mg |
复合益生菌苜蓿冻干原粉直接或与木糖醇等调配后,成型为片剂、粉剂和胶囊制剂中益生菌活菌数不低于2.0×109CFU/g。苜蓿益生菌片剂、粉剂和胶囊易于携带和使用、益生菌含量高、口感明显改善且贮藏稳定性好。富含蛋白质、膳食纤维等营养物质,此外还含有对人体有益的黄酮类、异黄酮类物质。L.plantarum SCI-01、L.plantarum P-8、B.lactis V9和L.caseiZhang发酵苜蓿嫩芽粉有利于维生素B6和B1的积累和活性异黄酮含量的提高。发酵后VB1含量较发酵前提高65.4%;发酵后活性较弱的葡萄糖苷型异黄酮(染料木苷)含量降低30.7%,而黄豆苷元的含量较发酵前提高了76.0%。
Claims (8)
1.一种复合益生菌发酵苜蓿嫩芽粉制剂,其特征在于,以植物乳杆菌SCI-01、植物乳杆菌P-8、动物双歧杆菌乳亚种V9和干酪乳杆菌Zhang为发酵菌剂,其保藏编号分别为CGMCCNo.11930、CGMCC No.6312、CGMCC No.5470和CGMCC No.5469;发酵培养基组成:以重量份数计,苜蓿嫩芽粉970-1000,蔗糖8-12,大豆蛋白粉8-12,料水比1:1;初始pH 6.0,接种上述四种菌剂,总接种量为1×107CFU/g,37℃发酵,pH 5.0为发酵终点;发酵产物经真空冷冻干燥获得复合益生菌发酵苜蓿嫩芽粉制剂。
2.如权利要求1所述的复合益生菌发酵苜蓿嫩芽粉制剂,其特征在于,所述发酵菌剂植物乳杆菌SCI-01、植物乳杆菌P-8、动物双歧杆菌乳亚种V9和干酪乳杆菌Zhang的接种量比例为1:1:1:2。
3.如权利要求1所述的复合益生菌发酵苜蓿嫩芽粉制剂,其特征在于,所述发酵产物与保护剂按照1:2的比例混合均匀后经真空冷冻干燥获得复合益生菌发酵苜蓿嫩芽粉制剂。
4.如权利要求3所述的复合益生菌发酵苜蓿嫩芽粉制剂,其特征在于,所述保护剂的组成为:海藻糖4%,乳糖4%,大豆蛋白粉2%,蒸馏水90%。
5.如权利要求2或4所述的复合益生菌发酵苜蓿嫩芽粉制剂,其特征在于,所述发酵产物或复合益生菌发酵苜蓿嫩芽粉制剂中的益生菌活菌数达到1.0×1010CFU/g以上。
6.权利要求1-5任一一项所述复合益生菌发酵苜蓿嫩芽粉制剂的用途,其特征在于,所述的苜蓿益生菌原粉制剂直接或与木糖醇调配后,制备成苜蓿益生菌片剂、粉剂或胶囊剂。
7.权利要求6所述复合益生菌发酵苜蓿嫩芽粉制剂的用途,其特征在于,经调配后的苜蓿益生菌片剂、粉剂或胶囊剂中的益生菌活菌数不低于2.0×109CFU/g。
8.一种复合益生菌发酵苜蓿嫩芽粉制剂的制备方法,其特征在于,以植物乳杆菌SCI-01、植物乳杆菌P-8、动物双歧杆菌乳亚种V9和干酪乳杆菌Zhang为发酵菌剂,其保藏编号分别为CGMCC No.11930、CGMCC No.6312、CGMCC No.5470和CGMCC No.5469;发酵培养基组成:以重量份数计,苜蓿嫩芽粉970-1000,蔗糖8-12,大豆蛋白粉8-12,料水比1:1;初始pH 6.0,按照1:1:1:2的比例接种上述四种菌剂,总接种量为1×107CFU/g,37℃发酵,pH 5.0为发酵终点;发酵产物与保护剂按照1:2的比例混合均匀后经真空冷冻干燥获得苜蓿益生菌原粉制剂;所述保护剂为:海藻糖4%,乳糖4%,大豆蛋白粉2%,蒸馏水90%;所述复合益生菌发酵苜蓿嫩芽粉制剂中的益生菌活菌数达到1.0×1010CFU/g以上。
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