CN114982893B - 魔芋菌解低聚糖速溶饮料的制备方法 - Google Patents
魔芋菌解低聚糖速溶饮料的制备方法 Download PDFInfo
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Abstract
本发明公开了一种魔芋菌解低聚糖速溶饮料的制备方法,利用微生物菌体嗜热脂肪芽孢杆菌及耐高温α‑淀粉酶的水解作用,将魔芋大分子葡甘聚糖水解成溶解性更好的较小分子的低聚糖,将魔芋中的淀粉等水不溶性物质水解为可溶性糖,再通过调配、干燥等加工工序,最终将不能直接食用、粘度大、口感差的魔芋粗粉加工成直接加水饮用、富含功能性魔芋低聚糖的固体速溶饮料。本发明制备的产品适口性好,饮用方便,对丰富魔芋加工品,扩大魔芋生产利用率有很好的作用。
Description
技术领域
本发明属于速溶饮料技术领域,具体涉及一种魔芋菌解低聚糖速溶饮料的制备方法。
背景技术
魔芋被认为是一种具有很好保健作用的食物资源。魔芋的功能性成分主要是葡甘聚糖,约占魔芋干物质总量的45%,在迄今已发现含有葡甘聚糖的植物材料中,魔芋的含量是最高的。葡甘聚糖不能被人体胃肠消化道中的酶水解,但作为一种低热量膳食纤维,其膨胀特性使其在胃肠道更易产生饱腹感觉,而且可以直接进入结肠,并被肠道微生物利用,在人体内发挥重要的保健作用。
富含葡甘聚糖的魔芋目前开发利用途径,主要是加工成魔芋豆腐、仿生凝胶食品,或提取作为食品增稠剂、凝固剂的葡甘聚糖等,其共同特征是利用葡甘聚糖的胶凝性制成凝胶固态、半固态食品。这类魔芋加工食品种类少,口味较单一,接受人群有限,不能很好发挥出魔芋及葡甘聚糖的功能作用和经济价值。将魔芋中的大分子葡甘聚糖通过降解变成小分子的低聚糖,使其水溶性和功能性增强,加工性能改善,利用率提高,这是魔芋行业发展的方向。现在诸多报道,魔芋葡甘聚糖的降解常使用的方法是化学酸水解和酶水解。酸水解效率高但存在食品安全性,酶水解专一性太强,且生产成本很高。
发明内容
本发明的目的是利用能降解魔芋葡甘聚糖成低分子的微生物,通过生物发酵,将不能直接食用、粘度大、口感差的魔芋粗粉加工成能直接加水饮用、富含功能性魔芋低聚糖的速溶饮料。
针对上述目的,本发明采用的技术方案由下述步骤组成:
1、制备嗜热脂肪芽孢杆菌菌液
将冻存的嗜热脂肪芽孢杆菌转接于试管斜面培养基上,在40~50℃培养36~48小时,进行活化;将活化后的嗜热脂肪芽孢杆菌转接于三角瓶液体培养基中,通过摇床振荡进行孢子培养,在50~55℃培养4~5天,摇床转速30~40转/分钟;待孢子培养好后,用无菌蒸馏水稀释液体培养基中的孢子数至105~106个/mL,得到嗜热脂肪芽孢杆菌菌液。
2、制备魔芋浆
将新鲜魔芋球茎去皮去芽后,迅速用打浆机破碎,用纯净水调节其可溶性固形物为10%~12%,再用胶体磨磨浆,使其为料水混合均匀、粘稠的魔芋浆;或者在干燥的魔芋粗粉中加入纯净水,充分搅拌均匀,获得魔芋粗粉浓度为90~120g/L的魔芋浆。
3、酸化脱腥和钝化酶
在魔芋浆中加入质量浓度为10%~12%的冰醋酸水溶液,冰醋酸的加入量为魔芋浆质量的1%~2%,用蒸汽夹层锅或加热罐迅速加热至75~85℃,在搅拌状态下保温15~20分钟,得到脱腥魔芋浆。
4、魔芋浆的菌解
按质量百分比组成为:魔芋浆83%~85%、大豆蛋白粉4%~6%、磷酸氢二钾0.25%~0.35%、余量为蒸馏水,在脱腥魔芋浆中加入大豆蛋白粉、磷酸氢二钾和水,混合均匀后用高压蒸汽杀菌锅在121℃灭菌15~22分钟,得到发酵液,然后迅速将发酵液冷却至50~60℃,转移到100L无菌发酵罐中,要求发酵罐带有内置搅拌,发酵罐搅拌器转速50~60转/分钟,发酵罐周围环境洁净度1万级;再向发酵罐中加入嗜热脂肪芽孢杆菌菌液和耐高温α-淀粉酶,用搅拌器充分搅拌分散均匀后停止搅拌,在50~55℃下静止发酵36~48小时,发酵结束后再次搅拌均匀发酵液。
5、调酸杀菌
向发酵液中加入冰醋酸,调节发酵液pH为3~3.5,在发酵罐中直接将发酵液加热至95~100℃,保温15~20分钟,将嗜热脂肪芽孢杆菌杀灭。
6、调配和干燥
向调酸杀菌后的发酵液中加入其质量0.018%~0.022%的甘草甜素和0.45%~0.55%的柠檬酸,用真空冷冻干燥机真空冷冻干燥或用喷雾干燥法干燥,得到速溶饮料。
上述步骤1中,优选所述斜面培养基的质量百分比组成为:牛肉浸出粉0.13%~0.15%、蛋白胨0.30%~0.40%、葡萄糖0.08%~0.12%、磷酸氢二钾0.15%~0.22%、氯化锰0.16%~0.24%、魔芋葡甘聚糖1.0%~1.5%、琼脂0.8%~1.0%、余量为蒸馏水,并用质量浓度为0.5%的氢氧化钠水溶液调节pH为7.2±0.2,121℃灭菌15~22分钟。
上述步骤1中,优选所述液体培养基的质量百分比组成为:牛肉浸出粉0.13%~0.15%、蛋白胨0.30%~0.40%、葡萄糖0.08%~0.12%、磷酸氢二钾0.15%~0.22%、氯化锰0.16%~0.24%、魔芋粗粉1.0%~1.4%、余量为蒸馏水,121℃灭菌15~22分钟。
上述步骤4中,优选所述嗜热脂肪芽孢杆菌菌液的接种量为发酵液质量的1.5%~2.5%,耐高温α-淀粉酶的加入量为发酵液质量的0.2%~0.25%,耐高温α-淀粉酶的酶活力为13万~16万U/mL。
上述步骤6中,所述真空冷冻干燥是先在-40~-50℃预冷冻,然后在-18℃真空冷冻干燥。所述喷雾干燥法干燥是用离心式喷雾干燥机干燥,进风温度160~170℃,出风温度为80~90℃,且干燥前先用蒸馏水调整发酵液中可溶性固形物在15%~20%。
本发明的有益效果如下:
本发明利用微生物菌体及酶的水解作用,将魔芋大分子葡甘聚糖水解成溶解性更好的较小分子的低聚糖,将魔芋中的淀粉等水不溶性物质水解为可溶性糖,再通过调配、干燥等加工工序,最终将魔芋全粉加工成功能性更强、水溶性很好的固体速溶饮料。本发明制备的产品适口性好,饮用方便,对丰富魔芋加工品,扩大魔芋生产利用率有很好的作用。
附图说明
图1是嗜热脂肪芽孢杆菌(Bacillus stearothermophilus)菌株形态,其中a是菌株在光学显微镜下的形态,b是菌株在基础培养基上的形态。
图2是菌解产物的薄层色谱分析。
图3是降解产物的MALDI-TOF-MS分析。
图4是不同浓度的魔芋低聚糖对DPPH·的清除率。
具体实施方式
下面结合附图和实施例对本发明进一步详细说明,但本发明的保护范围不仅限于这些实施例。
实施例1
1、制备嗜热脂肪芽孢杆菌菌液
将冻存的嗜热脂肪芽孢杆菌转接于试管斜面培养基上,在45℃培养42小时,进行活化。所述斜面培养基的质量百分比组成为:牛肉浸出粉0.15%、蛋白胨0.35%、葡萄糖0.1%、磷酸氢二钾0.2%、氯化锰0.2%、魔芋葡甘聚糖1.0%、琼脂1.0%、余量为蒸馏水,并用质量浓度为0.5%的氢氧化钠水溶液调节pH为7.2±0.2,121℃灭菌20分钟。
将活化后的嗜热脂肪芽孢杆菌转接于三角瓶液体培养基中,通过摇床振荡进行孢子培养,在55℃培养4天,摇床转速30转/分钟。所述液体培养基的质量百分比组成为:牛肉浸出粉0.15%、蛋白胨0.35%、葡萄糖0.1%、磷酸氢二钾0.2%、氯化锰0.2%、魔芋粗粉1.0%、余量为蒸馏水,121℃灭菌20分钟。
待孢子培养好后,用无菌蒸馏水稀释液体培养基中的孢子数至105~106个/mL,得到嗜热脂肪芽孢杆菌菌液。
2、制备魔芋浆
在干燥的魔芋粗粉中加入纯净水,充分搅拌均匀,获得魔芋粗粉浓度为110g/L的魔芋浆。
3、酸化脱腥和钝化酶
在魔芋浆中加入质量浓度为10%的冰醋酸水溶液,冰醋酸的加入量为魔芋浆质量的1.5%,用蒸汽夹层锅或加热罐迅速加热至80℃,在搅拌状态下保温20分钟,得到脱腥魔芋浆。
4、魔芋浆的菌解
按质量百分比组成为:魔芋浆85%、大豆蛋白粉5%、磷酸氢二钾0.3%、余量为蒸馏水,在脱腥魔芋浆中加入大豆蛋白粉、磷酸氢二钾和水,混合均匀后用高压蒸汽杀菌锅在121℃灭菌20分钟,得到发酵液,然后迅速将发酵液冷却至60℃,转移到100L无菌发酵罐中,要求发酵罐带有内置搅拌,发酵罐搅拌器转速50转/分钟,发酵罐周围环境洁净度1万级;再向发酵罐中加入嗜热脂肪芽孢杆菌菌液和耐高温α-淀粉酶,所述嗜热脂肪芽孢杆菌菌液的接种量为发酵液质量的2%,耐高温α-淀粉酶的加入量为发酵液质量的0.2%,耐高温α-淀粉酶的酶活力为15万U/mL。用搅拌器充分搅拌分散均匀后停止搅拌,在55℃下静止发酵48小时,发酵结束后再次搅拌均匀发酵液。
5、调酸杀菌
向发酵液中加入冰醋酸,调节发酵液pH为3~3.5,在发酵罐中直接将发酵液加热至95℃,保温20分钟,将嗜热脂肪芽孢杆菌杀灭。
6、调配和干燥
向调酸杀菌后的发酵液中加入其质量0.02%的甘草甜素和0.5%的柠檬酸,用真空冷冻干燥机先在-40℃预冷冻,然后在-18℃真空冷冻干燥,得到速溶饮料。
实施例2
1、制备嗜热脂肪芽孢杆菌菌液
将冻存的嗜热脂肪芽孢杆菌转接于试管斜面培养基上,在40℃培养48小时,进行活化。所述斜面培养基的质量百分比组成为:牛肉浸出粉0.13%、蛋白胨0.4%、葡萄糖0.12%、磷酸氢二钾0.22%、氯化锰0.16%、魔芋葡甘聚糖1.2%、琼脂1.0%、余量为蒸馏水,并用质量浓度为0.5%的氢氧化钠水溶液调节pH为7.2±0.2,121℃灭菌20分钟。
将活化后的嗜热脂肪芽孢杆菌转接于三角瓶液体培养基中,通过摇床振荡进行孢子培养,在50℃培养5天,摇床转速30转/分钟。所述液体培养基的质量百分比组成为:牛肉浸出粉0.13%、蛋白胨0.4%、葡萄糖0.12%、磷酸氢二钾0.22%、氯化锰0.16%、魔芋粗粉1.2%、余量为蒸馏水,121℃灭菌15分钟。
待孢子培养好后,用无菌蒸馏水稀释液体培养基中的孢子数至105~106个/mL,得到嗜热脂肪芽孢杆菌菌液。
2、制备魔芋浆
将新鲜魔芋球茎去皮去芽后,迅速用打浆机破碎,用纯净水调节其可溶性固形物为10%~12%,再用胶体磨磨浆,使其为料水混合均匀、粘稠的魔芋浆。
3、酸化脱腥和钝化酶
在魔芋浆中加入质量浓度为12%的冰醋酸水溶液,冰醋酸的加入量为魔芋浆质量的1%,用蒸汽夹层锅或加热罐迅速加热至85℃,在搅拌状态下保温15分钟,得到脱腥魔芋浆。
4、魔芋浆的菌解
按质量百分比组成为:魔芋浆83%、大豆蛋白粉4%、磷酸氢二钾0.25%、余量为蒸馏水,在脱腥魔芋浆中加入大豆蛋白粉、磷酸氢二钾和水,混合均匀后用高压蒸汽杀菌锅在121℃灭菌20分钟,得到发酵液,然后迅速将发酵液冷却至55℃,转移到100L无菌发酵罐中,要求发酵罐带有内置搅拌,发酵罐搅拌器转速50转/分钟,发酵罐周围环境洁净度1万级;再向发酵罐中加入嗜热脂肪芽孢杆菌菌液和耐高温α-淀粉酶,所述嗜热脂肪芽孢杆菌菌液的接种量为发酵液质量的2.5%,耐高温α-淀粉酶的加入量为发酵液质量的0.25%,耐高温α-淀粉酶的酶活力为13万U/mL。用搅拌器充分搅拌分散均匀后停止搅拌,在50℃下静止发酵48小时,发酵结束后再次搅拌均匀发酵液。
5、调酸杀菌
向发酵液中加入冰醋酸,调节发酵液pH为3~3.5,在发酵罐中直接将发酵液加热至95℃,保温20分钟,将嗜热脂肪芽孢杆菌杀灭。
6、调配和干燥
向调酸杀菌后的发酵液中加入其质量0.018%的甘草甜素和0.45%的柠檬酸,用真空冷冻干燥机先在-50℃预冷冻,然后在-18℃真空冷冻干燥,得到速溶饮料。
实施例3
1、制备嗜热脂肪芽孢杆菌菌液
将冻存的嗜热脂肪芽孢杆菌转接于试管斜面培养基上,在50℃培养36小时,进行活化。所述斜面培养基的质量百分比组成为:牛肉浸出粉0.15%、蛋白胨0.3%、葡萄糖0.08%、磷酸氢二钾0.15%、氯化锰0.24%、魔芋葡甘聚糖1.5%、琼脂0.8%、余量为蒸馏水,并用质量浓度为0.5%的氢氧化钠水溶液调节pH为7.2±0.2,121℃灭菌15分钟。
将活化后的嗜热脂肪芽孢杆菌转接于三角瓶液体培养基中,通过摇床振荡进行孢子培养,在55℃培养4天,摇床转速40转/分钟。所述液体培养基的质量百分比组成为:牛肉浸出粉0.15%、蛋白胨0.3%、葡萄糖0.08%、磷酸氢二钾0.15%、氯化锰0.24%、魔芋粗粉1.4%、余量为蒸馏水,121℃灭菌20分钟。
待孢子培养好后,用无菌蒸馏水稀释液体培养基中的孢子数至105~106个/mL,得到嗜热脂肪芽孢杆菌菌液。
2、制备魔芋浆
在干燥的魔芋粗粉中加入纯净水,充分搅拌均匀,获得魔芋粗粉浓度为90g/L的魔芋浆。
3、酸化脱腥和钝化酶
在魔芋浆中加入质量浓度为10%的冰醋酸水溶液,冰醋酸的加入量为魔芋浆质量的2%,用蒸汽夹层锅或加热罐迅速加热至75℃,在搅拌状态下保温20分钟,得到脱腥魔芋浆。
4、魔芋浆的菌解
按质量百分比组成为:魔芋浆85%、大豆蛋白粉6%、磷酸氢二钾0.35%、余量为蒸馏水,在脱腥魔芋浆中加入大豆蛋白粉、磷酸氢二钾和水,混合均匀后用高压蒸汽杀菌锅在121℃灭菌15分钟,得到发酵液,然后迅速将发酵液冷却至50℃,转移到100L无菌发酵罐中,要求发酵罐带有内置搅拌,发酵罐搅拌器转速60转/分钟,发酵罐周围环境洁净度1万级;再向发酵罐中加入嗜热脂肪芽孢杆菌菌液和耐高温α-淀粉酶,所述嗜热脂肪芽孢杆菌菌液的接种量为发酵液质量的1.5%,耐高温α-淀粉酶的加入量为发酵液质量的0.2%,耐高温α-淀粉酶的酶活力为16万U/mL。用搅拌器充分搅拌分散均匀后停止搅拌,在55℃下静止发酵36小时,发酵结束后再次搅拌均匀发酵液。
5、调酸杀菌
向发酵液中加入冰醋酸,调节发酵液pH为3~3.5,在发酵罐中直接将发酵液加热至100℃,保温15分钟,将嗜热脂肪芽孢杆菌杀灭。
6、调配和干燥
向调酸杀菌后的发酵液中加入其质量0.022%的甘草甜素和0.55%的柠檬酸,并用蒸馏水调整发酵液中可溶性固形物在15%~20%之间,然后用离心式喷雾干燥机干燥,进风温度160~170℃,出风温度为80~90℃。
为了确定本发明的制备方法,发明人进行了大量的实验室研究实验,具体实验情况如下:
1、魔芋葡甘聚糖降解菌株的分离和鉴定
(1)菌的分离和显微观察
随机选取罹患软腐病的魔芋球茎5个,每个魔芋分5点在球茎腐败部位取样,用75%(v/v)酒精对取样点表面消毒后,用接种环挑取腐败成分于含4%葡甘聚糖的基础培养基上,发现有些菌落对培养基有水解作用,出现半透明水浸斑。再将其菌转接到用魔芋粗粉制成的平板培养基上,37℃培养24~36 h,发现该菌株在魔芋粗粉制成的平板培养基上生长很好,将固态粘稠的培养基变成液态状。对该菌株进行革兰氏染色,在光学显微镜下观察结果,并对该菌株的部分生理生化性质进行检测,结果见图1,其生化特征见表1。从检测结果初步判断该菌株为芽孢细菌。
表1 分离菌株生理生化性质
注:“+”表示阳性反应;“-”表示阴性反应。
(2)菌株分子生物学鉴定
提取该菌株的基因组DNA,通过PCR进行16S rDNA的基因克隆。用正向引物5′-AGAGTTGATCMGGCTCAG-3′和反向引物5′-TACGGYTACCTTGTTACGATT-3′对菌株进行PCR扩增。将PCR扩增产物回收后进行测序。将测序结果提交至美国国家生物技术信息中心的GenBank核酸序列数据库中,通过基本局部比对搜索工具(BLAST)和CLUSTALX1.8软件菌株16SrDNA基因序列进行同源性比对,使用MEGA7.0版进行序列比对和比较,构建系统发育树。分析显示分离菌株为嗜热脂肪芽孢杆菌(Bacillus stearothermophilus)。
2、总氮对嗜热脂肪芽孢杆菌对水解魔芋粗粉的影响
无论用新鲜魔芋制成的魔芋浆还是用魔芋干粉制成的魔芋浆,蛋白质含量均很低,魔芋干粉中粗蛋白只有7g/100g左右,按照芽孢菌生长营养条件,魔芋浆是富碳少氮的培养基,因此应在发酵液中补加氮源,促进该芽孢菌的生长繁殖。
3、嗜热脂肪芽孢杆菌降解产物的分析
对菌解时间分别为0h、2h、4h、6h、8h、10h的魔芋粗粉降解产物进行TLC分析,观察随着菌解时间的延长,魔芋粗粉转化为低聚糖的变化情况,如图2所示。从图2显色的情况来看,在菌解时间为0~2h时,魔芋粗粉中含有少量在单糖与二糖之间的糖;菌解时间从2h延长至4h时,单糖与二糖之间的糖被嗜热脂肪芽孢杆菌逐渐利用,魔芋粗粉开始被降解为小分子的糖,当菌解时间继续延长至10h时,随着时间的延长,低聚糖的含量逐渐增加,表明高聚合度的魔芋葡甘聚糖含量正在降低。将处理48h所得发酵液在3000r/min离心10min,取上清液作为样品,随后使用MALDI-TOF MS进行分析。MALDI-TOF-MS用于确定经嗜热脂肪芽孢杆菌降解得到魔芋低聚糖的组成。图3的(A)和(B)显示了基于MALDI-TOF-MS信号高度的低聚糖组成,图3(C)为空白对照组。根据离子质量-电荷比(质荷比,m/z)值,对反应混合物的MALDI-TOF MS分析证实了嗜热脂肪芽孢杆菌水解魔芋葡甘聚糖产生一系列典型的低聚糖和少量相应的氧化低聚糖。进一步分析魔芋葡甘聚糖可完全降解为还原端为葡萄糖或者甘露糖的低聚糖,生成的低聚糖聚合度在2~14,主要产物为二糖和三糖。
4、魔芋菌解低聚糖对乳酸菌功能性评价
乳酸菌属作为公认的有益菌群,可以通过减少致病菌和提高免疫力来改善肠道微环境。但是由于胃酸的影响,乳酸菌的作用效果并不好。低聚糖对肠道乳酸菌的生长有促进作用。通过试验研究魔芋粗粉降解产物对于不同乳酸菌的增殖作用,同时比较魔芋低聚糖与魔芋粗粉和葡萄糖对乳酸菌的增殖效果,评价魔芋低聚糖的功能作用,结果如表2所示。
表2 魔芋水解低聚糖对乳酸菌的增殖作用
由表2可知,在乳酸菌培养时,魔芋低聚糖、葡萄糖、魔芋粗粉对植物乳杆菌体外增殖作用的光密度(OD)值在24h时分别为0.236、1.371和0.775,魔芋低聚糖、葡萄糖和魔芋粗粉对短乳杆菌体外增殖作用的OD值在24h时分别为0.224、1.339和0.791,魔芋低聚糖、葡萄糖和魔芋粗粉对干酪乳杆菌的体外增殖作用的OD值在24h时分别为0.243、1.336和0.916;可以看出,魔芋低聚糖对于三种乳酸菌的增殖效果介于魔芋粗粉和葡萄糖之间。通过pH值的结果可以看出,随着乳酸菌培养时间的延长,样品的pH值呈下降趋势。这些结果表明,由嗜热脂肪芽孢杆菌降解魔芋葡甘聚糖产生的魔芋低聚糖可能对调节肠道菌群平衡具有潜在作用。
5、魔芋菌解低聚糖抗氧化作用
不同浓度的魔芋粗粉菌解低聚糖对DPPH·的清除率如图4所示。如图4所示,在所选浓度范围内,随着魔芋粗粉和魔芋粗粉菌解低聚糖浓度的增加,对DPPH·的清除率均呈现逐渐加强的趋势,并且菌解后对DPPH·的清除率明显优于同浓度的魔芋粗粉溶液。当菌解后的魔芋粗粉低聚糖浓度为10 mg/mL时,对DPPH·的清除率达到最大,为81.43%。证明魔芋低聚糖对于DPPH·具有一定的清除作用,且随着分子量的降低,多糖及其衍生物的抗氧化活性将增强。
Claims (6)
1.一种魔芋菌解低聚糖速溶饮料的制备方法,其特征在于:
(1)制备嗜热脂肪芽孢杆菌菌液
将冻存的嗜热脂肪芽孢杆菌转接于试管斜面培养基上,在40~50℃培养36~48小时,进行活化;将活化后的嗜热脂肪芽孢杆菌转接于三角瓶液体培养基中,通过摇床振荡进行孢子培养,在50~55℃培养4~5天,摇床转速30~40转/分钟;待孢子培养好后,用无菌蒸馏水稀释液体培养基中的孢子数至105~106个/mL,得到嗜热脂肪芽孢杆菌菌液;
(2)制备魔芋浆
将新鲜魔芋球茎去皮去芽后,迅速用打浆机破碎,用纯净水调节其可溶性固形物为10%~12%,再用胶体磨磨浆,使其为料水混合均匀、粘稠的魔芋浆;或者在干燥的魔芋粗粉中加入纯净水,充分搅拌均匀,获得魔芋粗粉浓度为90~120g/L的魔芋浆;
(3)酸化脱腥和钝化酶
在魔芋浆中加入质量浓度为10%~12%的冰醋酸水溶液,冰醋酸的加入量为魔芋浆质量的1%~2%,用蒸汽夹层锅或加热罐迅速加热至75~85℃,在搅拌状态下保温15~20分钟,得到脱腥魔芋浆;
(4)魔芋浆的菌解
按质量百分比组成为:脱腥魔芋浆83%~85%、大豆蛋白粉4%~6%、磷酸氢二钾0.25%~0.35%、余量为蒸馏水,在脱腥魔芋浆中加入大豆蛋白粉、磷酸氢二钾和蒸馏水,混合均匀后用高压蒸汽杀菌锅在121℃灭菌15~22分钟,得到发酵液,然后迅速将发酵液冷却至50~60℃,转移到100L无菌发酵罐中,要求发酵罐带有内置搅拌器,发酵罐搅拌器转速50~60转/分钟,发酵罐周围环境洁净度1万级;再向发酵罐中加入嗜热脂肪芽孢杆菌菌液和耐高温α-淀粉酶,用搅拌器充分搅拌分散均匀后停止搅拌,在50~55℃下静止发酵36~48小时,发酵结束后再次搅拌均匀发酵液;
(5)调酸杀菌
向发酵液中加入冰醋酸,调节发酵液pH为3~3.5,在发酵罐中直接将发酵液加热至95~100℃,保温15~20分钟,将嗜热脂肪芽孢杆菌杀灭;
(6)调配和干燥
向调酸杀菌后的发酵液中加入其质量0.018%~0.022%的甘草甜素和0.45%~0.55%的柠檬酸,用真空冷冻干燥机真空冷冻干燥或用喷雾干燥法干燥,得到速溶饮料。
2.根据权利要求1所述的魔芋菌解低聚糖速溶饮料的制备方法,其特征在于:步骤(1)中,所述斜面培养基的质量百分比组成为:牛肉浸出粉0.13%~0.15%、蛋白胨0.30%~0.40%、葡萄糖0.08%~0.12%、磷酸氢二钾0.15%~0.22%、氯化锰0.16%~0.24%、魔芋葡甘聚糖1.0%~1.5%、琼脂0.8%~1.0%、余量为蒸馏水,并用质量浓度为0.5%的氢氧化钠水溶液调节pH为7.2±0.2,121℃灭菌15~22分钟。
3.根据权利要求1所述的魔芋菌解低聚糖速溶饮料的制备方法,其特征在于:步骤(1)中,所述液体培养基的质量百分比组成为:牛肉浸出粉0.13%~0.15%、蛋白胨0.30%~0.40%、葡萄糖0.08%~0.12%、磷酸氢二钾0.15%~0.22%、氯化锰0.16%~0.24%、魔芋粗粉1.0%~1.4%、余量为蒸馏水,121℃灭菌15~22分钟。
4.根据权利要求1所述的魔芋菌解低聚糖速溶饮料的制备方法,其特征在于:步骤(4)中,所述嗜热脂肪芽孢杆菌菌液的接种量为发酵液质量的1.5%~2.5%,耐高温α-淀粉酶的加入量为发酵液质量的0.2%~0.25%,耐高温α-淀粉酶的酶活力为13万~16万U/mL。
5.根据权利要求1所述的魔芋菌解低聚糖速溶饮料的制备方法,其特征在于:步骤(6)中,所述真空冷冻干燥是先在-40~-50℃预冷冻,然后在-18℃真空冷冻干燥。
6.根据权利要求1所述的魔芋菌解低聚糖速溶饮料的制备方法,其特征在于:步骤(6)中,所述喷雾干燥法干燥是用离心式喷雾干燥机干燥,进风温度160~170℃,出风温度为80~90℃,且干燥前先用蒸馏水调整发酵液中可溶性固形物在15%~20%。
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