CN104643246A - 一株青春双歧杆菌在制备活性双歧杆菌发酵饮料中的应用 - Google Patents
一株青春双歧杆菌在制备活性双歧杆菌发酵饮料中的应用 Download PDFInfo
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- CN104643246A CN104643246A CN201510109572.8A CN201510109572A CN104643246A CN 104643246 A CN104643246 A CN 104643246A CN 201510109572 A CN201510109572 A CN 201510109572A CN 104643246 A CN104643246 A CN 104643246A
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/38—Other non-alcoholic beverages
- A23L2/382—Other non-alcoholic beverages fermented
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/51—Bifidobacterium
- A23V2400/513—Adolescentes
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- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
本发明公开了一株青春双歧杆菌在制备发酵沙棘饮料中的应用。所采用青春双歧杆菌(Bifidobacterium adolescentis)BL-8CGMCC No.7791,是一株具有广谱抑菌效果的产细菌素双歧杆菌,且具有很强的耐消化道逆环境、生物拮抗和抗氧化能力,可用于制备活菌发酵饮料。本发明制备的双歧杆菌发酵沙棘汁活菌饮料色泽温和(图1),口感好,风味佳,活菌数高,货架期较长,是具有良好保健效果的活菌饮品。
Description
技术领域
本发明涉及一株双歧杆菌在制备活性双歧杆菌发酵饮料中的应用,特别涉及一株青春双歧杆菌菌株发酵生产沙棘汁活菌饮料的工艺。
背景技术
乳酸菌饮料可促进人体肠道消化吸收,维护人体肠道微生态平衡,抑菌抗病,增强人体免疫系统等保健功效。随着人们对发酵乳营养价值认识及消费能力的提高,“健康”概念势必将引领乳酸菌饮料成为乳制品市场中另一个消费热点。
双歧杆菌(Bifidobacterium spp.)是乳酸菌的一个重要分支,因其调节肠道平衡,提高免疫力等众多生理功能受到大众的关注。目前,国内外已开发的乳酸菌饮料中,有少数也采用双歧杆菌,如动物双歧杆菌(Bifidobacterium animalis)、长双歧杆菌(Bifidobacterium longum)等,还未见任何青春双歧杆菌(Bifidobacterium adolescentis)包括产细菌素菌株发酵活性饮料的专利及文献报道。乳酸菌细菌素是乳酸菌在代谢过程中通过核糖体合成机制产生的具有抑菌活性的多肽或蛋白类物质,可抑制或杀死部分食品腐败菌和致病菌的生长,具有耐受一定热和酸碱以及在人体内可降解、无毒无残留的特点。且双歧杆菌在许多方面对人体健康起到了不可替代的作用,如保持和恢复人体正常肠道菌群的平衡,并刺激肠道蠕动,有免疫调节、延缓衰老、抗癌及抗肿瘤等生理作用。其能经受住胃酸和胆汁酸的环境存活下来并在人肠道内定植,发挥对人体有益的及重要生理作用。以产细菌素的双歧杆菌发酵生产活菌饮料,不仅会极大增强其保健及生理调节功能,提高其附加值;而且可以明显延长产品的货架期,保证产品安全性。
沙棘学名(Hippophae rhamnoides Linn),又名醋柳、墨刺,是沙棘为药食同源植物,果实含有丰富的营养物质和生物活性物质。沙棘果实中维生素C极高,同时含有人体8种必需氨基酸及大量的非蛋白氮,可在一定程度促进双歧杆菌的生长。
以双歧杆菌发酵沙棘汁制成活菌饮料,可同时发挥沙棘汁的营养及生理功效和双歧杆菌的保健作用,必将成为高附加值的活菌饮品。
发明内容
本发明的目的是提供一种青春双歧杆菌发酵沙棘汁活性饮料的制备方法。
青春双歧杆菌(Bifidobacterium adolescentis)BL-8 CGMCC №.7791已于2013年06月21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,地址为: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),保藏登记号为:CGMCC №.7791。
本发明用于发酵的青春双歧杆菌BL-8 CGMCC №.7791是一株具有广谱抑菌效果的产细菌素双歧杆菌,其所产细菌素不仅对李斯特菌、葡萄球菌、芽孢杆菌等革兰氏阳性菌有明显抑制作用,对大肠杆菌等阴性菌也表现出非常强烈的抑菌活性。同时,该菌株具有耐受消化道逆环境、生物拮抗及抗氧化等保健功效,可用于制备双歧杆菌活菌饮料。
本发明提供一种青春双歧杆菌发酵沙棘汁活性饮料的制备方法,即将青春双歧杆菌(Bifidobacterium adolescentis)BL-8 CGMCC №.7791活化后接种在含有沙棘汁、乳粉和蔗糖复合溶液的培养基中,在一定温度下厌氧培养一定时间后,结束发酵,获得双歧杆菌发酵沙棘汁活菌饮料。
所述方法中,沙棘汁发酵培养基按下述方法制备:取新鲜沙棘和质量比为1∶1的水混合打浆,用四层纱布过滤,得到质量百分数为50%的沙棘汁,并用Na2CO3调节其pH至6.0左右;将蔗糖100g溶于100mL水中得到蔗糖溶液;在100mL水中加入100g脱脂乳粉,充分搅拌溶解,得脱脂复原乳。按照沙棘汁56%、蔗糖溶液16%、脱脂复原乳9%、水19%配制,并用均质机在8MPa条件下进行第一次均质,12MPa条件下进行第二次均质,得到复合果汁发酵培养基;最后,在复合果汁发酵培养基里充氮,使氧还电位降至-0.03eV以下,121℃、10min灭菌。
所述方法中,青春双歧杆菌(Bifidobacterium adolescentis)BL-8 CGMCC №.7791的培养时间为24-26h,接种量为1×104cfu/mL,培养温度为37℃。
本发明利用上述青春双歧杆菌,以极具营养保健价值的沙棘为发酵原料,同时添加蔗糖及脱脂乳,制备了一种青春双歧杆菌发酵沙棘汁活菌饮料。原料经该双歧杆菌菌株发酵后可产生乙酸、乳酸、B族维生素和氨基酸类等具有重要生理活性的代谢产物,同时具有良好的感官及营养品质。发酵成品在4℃和室温条件下分别可贮藏21天和7天。本发明制备的双歧杆菌发酵沙棘汁活菌饮料色泽温和(图1),酸甜适口且活菌数高、货架期长,是具有良好营养保健效果和感官品质的保健饮品。
附图说明
图1为本发明的沙棘汁双歧杆菌发酵饮料样品(包装形式之一)。
图2为沙棘汁双歧杆菌发酵饮料分别在4℃和室温下厌氧保藏时双歧杆菌活菌数随时间的变化。
具体实施方式
下述实施例中所述实验方法,如无特殊说明,均为常规方法。
实施例1、青春双歧杆菌(Bifidobacterium adolescentis)BL-8的功效评价
青春双歧杆菌(Bifidobacterium adolescentis)BL-8来源于新疆吐鲁番地区长寿老人粪便,并已于2013年06月21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,地址为: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),保藏登记号为:CGMCC №.7791。
1、耐消化道逆环境试验
模拟人体胃肠道环境,对BL-8耐消化道逆环境特性进行研究。
(1)酸耐受性试验:用37%的浓盐酸调节PBS(pH7.0)缓冲液pH至3.0。接入活化3代的双歧杆菌BL-8液体培养物,接种量5%,37℃厌氧培养2h。分别于0和2h取样,以亨盖特厌氧滚管法测定活菌数,计算存活率。
(2)胆汁酸盐耐受性试验:向改良MRS液体培养基中加入0.5%牛胆酸钠,121℃、20min灭菌,接入活化3代的双歧杆菌BL-8液体培养物,接种量2%,37℃厌氧培养12h。取样测定活菌数;同时以不含胆汁酸盐的改良MRS液体培养基为对照。计算存活率。所用改良MRS培养基的配制(以1000mL计):葡萄糖20g,牛肉浸粉10g,蛋白胨10g,酵母浸粉5g,玉米膏3g,无水乙酸钠5g,柠檬酸三胺2g,磷酸氢二钾2g,硫酸锰0.25g,硫酸镁0.58g,L-半胱氨酸盐酸0.4g,吐温-801mL。
(3)耐胃蛋白酶试验:向PBS(pH7.0)缓冲液中加入37%的浓盐酸,调节其pH至3.0,然后加入0.5%胃蛋白酶作为模拟胃液,接入菌种活化3代后的BL-8液体培养物,接种量5%,于0和2h取样测定活菌数,计算存活率。
(4)耐胰蛋白酶试验:向PBS(pH7.0)缓冲液中加入1%胰蛋白酶作为模拟肠液,按5%的接菌量无菌接种,37℃厌氧培养12h,分别于0h和12h取样测定活菌数,计算存活率。
表1 不同处理对菌株BL-8存活率的影响
从表1分析结果可以看出,菌株BL-8经酸处理后存活率为93.2%,胆汁酸盐及胃蛋白酶处理后存活率均在87%以上,胰蛋白酶处理12h后菌株有所增长,说明双歧杆菌BL-8对酸、胆汁酸盐、胃蛋白酶及胰蛋白酶均有很强的耐受力,确定菌株BL-8可用于开发微生态制剂及其它产品。
2、菌体细胞的生物拮抗作用分析
按1%接种量向改良MRS液体培养基中接种受试指示菌(见表2)和青春双歧杆菌BL-8液体培养物,混匀后37℃厌氧培养24h,取样计数。采用各指示菌相应选择性培养基,通过平板计数法考察各指示菌的存活情况。同时以受试指示菌在改良MRS培养基中的纯培养物为对照。其中,金黄色葡萄球菌(Staphylococcus aereu)所用选择性培养基为Baird-Parker培养基,李斯特菌(Listeria spp.)用李斯特氏菌显色培养基,大肠杆菌(Escherichia coli)用伊红美蓝培养基,芽孢杆菌(Bacillus spp.)用甘露醇卵黄多粘菌素琼脂基础培养。结果表明,各指示菌对照组在MRS液体培养基中生长态势良好,培养24h后活菌数对数值均在8以上。与青春双歧杆菌BL-8混合培养后的结果显示各指示菌的生长均受到抑制。其中,大肠杆菌1.90、金黄色葡萄球菌1.128、金黄色葡萄球菌1.169以及凝结芽孢杆菌的活菌数下降明显(1.59-2.07log),生长受到明显抑制。以上结果说明,双歧杆菌BL-8对8株受试菌均表现不同程度的拮抗作用,这可能与受试菌和双歧杆菌BL-8之间的营养竞争或空间竞争有关,以及BL-8在生长过程产生有机酸和细菌素有关。
表2 菌株BL-8菌体细胞的生物拮抗作用
注:NICPBP为卫生部药品生物制品检定,ATCC为美国标准菌种保藏所,CGMCC为普通微生物菌种保藏管理中心。
综上所述,菌株BL-8的菌体细胞对常见致病菌表现出很强的生物拮抗作用,将其用于制备活菌发酵饮料,对饮料安全性会有很大程度提高。
3、BL-8的抗氧化功效评价
通过超氧阴离子自由基清除实验,DPPH自由基清除实验,羟自由基清除实验以及还原能力测定实验,对菌株BL-8的菌体细胞、菌体无细胞提取物以及无细胞发酵上清液的抗氧化能力进行研究。
(1)样品的制备:样品为菌体细胞、菌体无细胞提取物以及无细胞发酵上清液。其制备方法入下:将青春双歧杆菌BL-8于37℃厌氧培养24h。将发酵液以5000rpm/min离心15min,收集上清液,得到无细胞发酵上清液样品;同时收集菌体细胞,并用双蒸水洗涤3次,最后用双蒸水将其菌数调整到109cells/mL,所得菌悬液分为两组,其中一组作为菌体细胞样品;将菌悬液的另外一组放于冰浴中超声破碎,并将破碎液于6000rpm/min离心10min,收集上清液,即为菌体无细胞提取物样品。
(2)清除超氧阴离子自由基试验:采用邻苯三酚自氧化法测定其抗氧化能力。向Tris-HCl(4.5mL、0.1mol/L)缓冲液(pH8.2)中依次加入乙二胺四乙酸(热水溶解,1mL、1.0mmol/L)、样品(1.0mL)、蒸馏水(2.4mL),25℃反应10min,然后加入邻苯三酚2mL,25℃反应60min,接着加入HCl(100μL、12mol/L)终止反应。在波长325nm处测定吸光度(AS)。空白管以1.0mL蒸馏水代替样品,操作方法同样品管,测得吸光度(AC)。平行测定3次取平均值。其中,0.1mol/L、pH 8.2的Tris-HCl的制备如下:Tris碱0.346g,1000ml容量瓶定容,用浓盐酸调其pH至8.2。清除率通过以下公式计算:清除率/%=[(Ac-As)/Ac]×100
(3)清除DPPH自由基试验:向4.0mL、0.1mmol/L DPPH溶液中加入样品2.0mL和50%乙醇1.0mL,在暗处反应60min。反应结束后将反应液以12000rpm/min离心20min,取其上清液,波长517nm处测吸光度(Aj)。空白管以2.0mL蒸馏水代替样品,操作方法同样品管,测得吸光度(Ai)。平行测定3次取平均值。清除率通过以下公式计算:清除率/%=[1-Ai/A]×100
(4)清除羟自由基试验:向lmL pH为7.4,浓度为0.05mol/L的PBS缓冲液中加入6mmol/L的邻菲罗林0.5mL,混匀,快速加入0.5mL的FeSO4(6mmol/L)溶液并混匀。加入0.5mL样品溶液并混匀,再加入0.5mL的H2O2(0.1%),最后用蒸馏水将体积补充到4mL。同时做空白对照实验。另再做损伤管和未损伤管,其中,损伤管中加入0.5mL、0.1%的H2O2,不加样品。未损伤管不加H2O2,也不加样品。最后均将体积补充到4mL。37℃保温1h后测反应液在536nm波长处的吸光度。清除率通过以下公式计算:清除率/%=[(Ai-A)/(A0-A)]×100,A为不加样品时溶液的吸光度;A0为不加样和H2O2时溶液的吸光度;Ai为加样品时溶液的吸光度。
(5)抗脂质过氧化率的检测:向0.5mL PBS溶液(0.02mol/L,pH7.4)中加入1mL亚油酸的乳化液、1%FeSO4 1mL后加入0.5mL样品,37℃水浴反应1.5h,向混合液中加入0.2mL4%TCA(三氯乙酸)、2mL 0.8%TBA(硫代巴比妥酸),然后将反应液于100℃水浴反应30min,迅速冷却后离心、纱布过滤,收集上清液,在波长532nm处测吸光值。空白对照组以0.5mL无菌生理盐水代替样品液,以PBS液加等体积的样品液离心、过滤后调零。硫代巴比妥酸溶解:(加约20ml 1mol/L的NaOH溶液,在60℃超声溶解4h左右)
清除率/%=[1-Ai/A]×100
(6)还原能力测定:向浓度0.2mol/L、pH6.6的磷酸缓冲溶液0.5mL及1g/100mL铁氰化钾0.5mL中加入0.5mL样品,于50℃水浴20min后迅速冷却。再加入10g/100mL三氯乙酸0.5mL,3000rprn/min离心5min,取上清液1mL,加入1mL蒸馏水及1mL 0.1g/100mL三氯化铁,混合均匀。10min后,于700nm波长测定其吸光度。吸光度越大表示待测样品的还原能力越强。
表3 菌株BF-10的抗氧化活性分析
结果表明:对超氧阴离子清除率最好的是菌体无细胞提取物菌体细胞,清除率为63.48%,说明清除超氧阴离子的活性物质主要是青春菌体细胞的胞内物质;菌体细胞、无细胞发酵上清液和菌体无细胞提取物对DPPH的清除率相当,分别为71.38%、71.51%和76.38%,说明不论菌体细胞、无细胞发酵上清液还是菌体无细胞提取物均含有较丰富的清除DPPH自由基的活性物质;菌体细胞对羟自由基清除率达到71.08%,且明显高于菌体无细胞提取物和无细胞发酵上清液对其清除率,说明清除羟自由基的活性物质主要是说明主要存在于青春双歧杆菌BL-8的菌体细胞表面;同样可得抗脂质过氧化的活性物质主要是婴儿菌体细胞的胞内物质,还原能力的活性物质主要存在于菌体细胞胞外。本研究结果显示,菌株BL-8的菌体细胞及胞外分泌物均具有较强的抗氧化能力。
综上所述,青春双歧杆菌BL-8对消化道逆环境的耐受力很强,且对多种食源性致病菌及腐败菌有抑制作用,同时其菌体细胞和胞外分泌物对自由基具有很好的清除作用、抗脂质过氧化及还原能力高,有作为益生菌菌株应用以及开发系列产品的潜力。
实施例2、青春双歧杆菌(Bifidobacterium adolescentis)BL-8制备沙棘汁发酵饮料
本发明利用青春双歧杆菌(Bifidobacterium adolescentis)BL-8制备沙棘汁双歧杆菌发酵饮料,制备方法如下所述:
1、制备双歧杆菌种子
(1)一代种子的制备:向改良MRS液体培养基中接种青春双歧杆菌(Bifidobacteriumadolescentis)BL-8,37℃厌氧培养24-48h,得到一代种子。
(2)二代种子的制备:将一代种子接种在改良MRS液体培养基中37℃厌氧培养14-16h,得到第二代种子。
(3)三代种子的制备:将二代种子接种在改良MRS培养基中厌氧培养14h,得到第三代种子。将第三代种子作为培养种子使用。
2、沙棘汁发酵培养基的制备
(1)沙棘汁的获得:取新鲜沙棘和质量比为1∶1的水混合打浆,用四层纱布过滤,得到质量百分数为50%的沙棘汁,并用碱面(Na2CO3)调节其pH至6.0左右;
(2)脱脂复原乳的制备:在100mL水中加入100g脱脂乳粉,充分搅拌溶解,得脱脂复原乳;
(3)蔗糖溶液的制备:将蔗糖100g溶于100mL水中得到蔗糖溶液;
(4)按照沙棘汁56%,蔗糖溶液16%,脱脂复原乳9%,水19%配制,并用均质机在8MPa条件下进行第一次均质,12MPa条件下进行第二次均质,得到复合果汁发酵培养基;
(5)在复合果汁发酵培养基里充氮,使氧还电位降至-0.03eV以下,121℃、10min灭菌。
3、制备双歧杆菌发酵沙棘汁活菌饮料
将步骤1活化三代得到的种子接种至含沙棘汁培养基中,接种量1%(1×104cfu/mL),37℃厌氧培养约24-26h,结束发酵,得到沙棘汁双歧杆菌发酵饮料。亨盖特厌氧滚管法计数活菌数达9.4×109cfu/mL。发酵饮料如图1所示,于4℃储存。
实施例3、青春双歧杆菌(Bifidobacterium adolescentis)BL-8制备沙棘汁发酵饮料的品质及货架期评价
1、品质评价
对发酵饮料进行感官评价和理化指标测定,分析活性双歧杆菌发酵饮料的品质。
(1)感官评价:对实施例2制备的双歧杆菌发酵沙棘汁活性饮料采用模糊数学七度标度法进行感官分析,分析方法如下:由经过培训的10人组成评分小组,评语论域为V=(1,2,3,4,5,6,7),1:最难接受;2:较难接受;3:稍难接受;4:勉强接受;5:较易接受;6:容易接受;7:最易接受。发酵果汁的评定领域U=(色泽,香气,酸度,甜度,苦涩味,组织状态),其对应的权重值为X=(0.15,0.20,0.20,0.15,0.20,0.10)。结果见表4,可以看出该饮料有沙棘果香及双歧杆菌发酵后特有的的芳香,其色泽、组织状态、甜度得分均在6.0以上。其综合感官得分为6.12,是一种容易被大众接受的饮品。
表4 发酵饮料感官评价得分表
(2)理化指标测定:蛋白质含量:参照GB 50095-2010《食品中蛋白质的测定》;可溶性固形物含量:参照GB 12143.1-88《软饮料中可溶性固形物的测定方法》;还原糖含量:参照GB/T 5009.7-2003《食品中还原糖的测定》;总糖含量:高效液相色谱法;脂肪含量:参照GB/T 5009.6-2003《食品中脂肪的测定方法》;总酸含量:参照GB/T 12456-2008《食品中总酸的测定方法》(以乳酸计)。结果见表5,可以看出,实施例2所制备活菌饮料的理化指标均符合国家标准GB 16321-2003《乳酸菌饮料卫生标准》的要求。
表5 发酵饮料的理化指标
2、货架期评价
分别于4℃和室温(22±1℃)保藏活菌饮料,每隔24h取样,亨盖特厌氧滚管法计数。结果如附图2可以看出,4℃贮藏过程中活菌饮料中双歧杆菌活菌数量下降缓慢,在21天内双歧杆菌活菌数仍保持在1.0×106cfu/mL(GB 16321-2003《乳酸菌饮料卫生标准》对乳酸菌饮料中乳酸菌活菌数的最低限量)以上;在室温贮藏过程中,饮料活菌数迅速下降,第8天活菌数已低于国标最低限量。以上表明,贮藏温度对该活菌饮品的货架期有显著影响,作为一种不经过二次杀菌的货架期较短的活菌饮品,该发酵饮料更适宜采取冷链销售模式。
Claims (6)
1.一种生产双歧杆菌发酵沙棘汁活菌饮料的方法,是用产细菌素青春双歧杆菌(Bifidobacterium adolescentis)BL-8 CGMCC7791发酵沙棘汁得到的活菌饮料。
2.根据权利要求1所述的方法,其特征在于:所述发酵沙棘汁培养基按下述方法制备:取新鲜沙棘和质量比为1∶1的水混合打浆,用四层纱布过滤,得到质量百分数为50%的沙棘汁,并用碱面(Na2CO3)调节其pH之6.0左右;将蔗糖100g溶于100mL水中得到蔗糖溶液;在100mL水中加入100g脱脂乳粉,充分搅拌溶解,得脱脂复原乳。按照沙棘汁56%,蔗糖溶液16%,低脂复原乳9%,水19%配制,并用均质机在8MPa条件下进行第一次均质,12MPa条件下进行第二次均质,得到复合果汁发酵培养基;最后,在复合果汁发酵培养基里充氮,使氧还电位降至-0.03eV以下,121℃、10min灭菌。
3.根据权利要求2所述方法,其特征在于:所述沙棘汁为质量百分数为50%的沙棘汁。
4.根据权利要求3所述方法,其特征在于:培养基组成为沙棘汁56%,蔗糖溶液16%,低脂复原乳9%,水19%;所述百分含量均为体积百分数。
5.根据权利要求1或2或3或4所述方法,其特征在于:所述培养时间为24-26h,接种量为1×104cfu/mL,培养温度为37℃。
6.权利要求1-5中任意一项所述的方法制备的双歧杆菌发酵沙棘汁活菌饮料。
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