CN117243982B - 一种利用酵母菌消除银杏外种皮臭味的方法 - Google Patents

一种利用酵母菌消除银杏外种皮臭味的方法 Download PDF

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CN117243982B
CN117243982B CN202311535342.9A CN202311535342A CN117243982B CN 117243982 B CN117243982 B CN 117243982B CN 202311535342 A CN202311535342 A CN 202311535342A CN 117243982 B CN117243982 B CN 117243982B
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张泓泰
慕钰文
杨新宇
杨建斌
王嫩寒
代小伟
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Abstract

本发明涉及一种利用酵母菌消除银杏外种皮臭味的方法,包括以下步骤:将成熟的银杏果实的外种皮和水打浆,制成浆液,将活化的酵母菌A接种于浆料,厌氧发酵;所述酵母菌A保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏编号为CGMCC NO.28245。本发明通过筛选,得到了一种酵母菌A,其接种于银杏外种皮浆料中,厌氧发酵后,可以明显减轻、消除银杏外种皮的臭味,且对银杏外种皮的其他有益活性成分基本没有影响和破坏,为银杏外种皮的加工,研究提供了工业上可行的基础。拓宽了银杏作物的利用价值。

Description

一种利用酵母菌消除银杏外种皮臭味的方法
技术领域
本发明属于酵母菌技术领域,特别涉及一种利用酵母菌消除银杏外种皮臭味的方法。
背景技术
银杏外种皮俗称白果“衣子”,肉质肥厚,约占种子总重的75%,尽管研究发现,银杏外种皮中含有氢化银杏酸、银杏酚、黄酮类、银杏甲素、银杏乙素和多糖类化合物等有效成分。银杏外皮的粗提物,具有抗结核分枝杆菌等抑菌和杀菌作用、抗炎、提高免疫力、清除自由基、镇咳祛痰、抗衰老和诱导癌细胞凋亡等多种药用价值。因此银杏外皮是一种重要的天然产物资源。但是银杏外皮有一种非常强烈并且令人不愉快的臭味,这主要是因为银杏外皮含有许多挥发性物质,比如丁酸、庚酸等低级脂肪酸。这些物质也存在于变质的奶酪、人体汗液中,挥发产生的味道令人不悦,甚至产生难以抑制的生理不适感,比如恶心,呕吐,眩晕等。因此成熟的外种皮气味臭味极强、有腐蚀性,银杏外种皮在在后续生产加工受到很大的限制,被当作废弃物丢掉,污染环境。
目前消除臭味的方法是剥皮后,用开水烫外皮,但是一方面扔具有大量的臭味产生,二是高温条件下对银杏外皮中需要的活性成分可能产生结构破坏,丧失或者改变了其生物活性。现有技术中还缺乏一种行之有效的,在不影响银杏外皮活性成分的情况下,减轻或者消除其臭味的方法。
发明内容
为了克服现有技术中还缺乏有效在避免对银杏外皮活性成分造成破坏的情况下消除其臭味,本发明提供了一种利用酵母菌消除银杏外种皮臭味的方法。具体而言,本发明筛选得到一种酵母发酵菌,通过基于发酵技术的加工方法后,通过嗅觉的气味评价和“液质”联用进行评估鉴定,确认基于使用该酵母菌基于发酵技术处理银杏外种皮,可以达到臭味消除,并且外种皮中的活性物质不影响的加工利用的目的。本发明通过以下技术手段达到上述目的:
一种利用酵母菌消除银杏外种皮臭味的方法,包括以下步骤:将成熟的银杏果实的外种皮和水打浆,制成浆液,将活化的酵母菌A接种于浆料,厌氧发酵;所述酵母菌A保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏编号为CGMCC NO. 28245。
进一步地,打浆时,银杏果实的外种皮和水的质量比为3-5:2。打浆的工艺参数没有特别的限定,采用打浆机将外种皮和水混合为均匀的浆料即可。比如在本发明一个具体实施方式中,打浆机的转速为400-600rpm,打浆时间为15-30min。
进一步地,酵母菌活化是将酵母菌A混入3-6wt%的白砂糖水中,酵母菌A浓度为1×103-5×103cfu/mL,然后在37-38℃的条件下培养,直到观察到液面有气泡生成,降至常温,此时的溶液即为经过活化的酵母菌A溶液。
进一步地,酵母菌接种浓度为50-200 cfu/mL,优选为100-150cfu/mL。
进一步地,加入水使浆料的可溶性固形物含量为10-15%,加入有机酸使浆料的pH为3-4,优选地,为3-3.3。所述有机酸选自柠檬酸、苹果酸、酒石酸、草酸中的至少一种。发明人发现,本发明筛选得到的酵母菌,对发酵体系的pH较为敏感,在pH为3-3.3时达到最佳的发酵效果。
进一步地,所述厌氧发酵是在20-30℃厌氧条件下发酵10-20天。优选地,在25±2℃发酵12-15天。
采用本发明酵母菌A发酵后的银杏外种皮浆料,已经基本消除了银杏外种皮的臭味,同时银杏外种皮中活性成分基本没有收到影响。本发明技术为银杏外种皮加工技术提供了便利,消除由于外种皮臭味带来的加工不便,为银杏外种皮工业化加工提供便利。
在本发明一个优选技术方案中,所述利用酵母菌A消除银杏外种皮臭味的方法,包括以下步骤:
(S1)收集无明显腐烂或损伤的成熟银杏果;
(S2)分拣出的银杏果进行清洗,脱除银杏果外种皮;
(S3)银杏果外种皮置于匀浆机中,按照银杏果:清水质量比3-5:2加入清水,将外种皮和水打浆得到均匀的浆状物;
(S4)加入清水调整匀浆的可溶性固形物含量,调节pH为3-4,将活化好的酵母菌A接种于匀浆中,然后20-30℃下厌氧静置发酵10-20天,发酵结束后,取出发酵后的银杏外种皮匀浆;所述酵母菌A保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏编号为CGMCC NO. 28245。
进一步地,步骤(S4)中,调整匀浆的可溶性固形物含量为10-15%,调节pH至3-3.3。
本发明还提供了一种用于消除银杏外种皮臭味的酵母菌A,保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏编号为CGMCC NO. 28245。
本发明还提供了一种酵母菌A在消除植物臭味中的用途,所述酵母菌,保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏编号为CGMCC NO. 28245。
进一步地,所述植物选自银杏。
本发明上述用于消除银杏外种皮臭味的酵母菌,通过包括以下步骤的方法得到:
从黄杏子自然发酵液中分离和筛选得到具有异味消除能力的酵母菌A。
更为具体地,酵母菌,通过包括以下步骤的方法得到:取杏果自然发酵液,用无菌水稀释0.5×106-1×106倍,得到稀释液,将稀释液均匀涂布在YPD培养基上,28℃恒温箱中培养48小时,选取具有典型酵母特征的单菌落(典型酵母特征是圆形、光滑、有光泽,大小中等,颜色为白色,质地湿润且膏状,且在显微镜下细胞呈椭圆形或圆形),接种到液体培养基中培养,分离出不同的酵母菌株,通过在培养基中添加银杏外种皮选择培养和筛选后,筛选并发现酵母菌株A在接种于银杏外种皮匀浆后,发酵液中银杏外种皮臭味消除明显,将该酵母菌命名为酵母菌A,进行菌种保藏。而其他菌株对异味消除不明显。
本发明还提供了一种以银杏外种皮为原料生产乙醇的方法,包括以下步骤:将成熟的银杏果实的外种皮和水打浆,制成浆液,将活化的酵母菌A接种于浆料,厌氧发酵,蒸馏提取乙醇;所述酵母菌A保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏编号为CGMCC NO. 28245。
通过蒸馏的方法将发酵后的混合物加热,使得其中的乙醇蒸发并通过冷凝管收集,得到纯度较高的乙醇。
发明人还发现,在利用本发明酵母菌A对银杏外种皮厌氧发酵过程中,不仅在不影响银杏外种皮活性成分含量下消除了臭味,而且发酵结束后,体系内产生了5-7wt%的酒精,可以分离回收利用。
本发明通过筛选,意外得到了一种酵母菌A,其接种于银杏外种皮浆料中,厌氧发酵后,可以明显减轻,消除银杏外种皮的臭味,且对银杏外种皮的其他有益活性成分基本没有影响和破坏。为银杏外种皮的加工,研究提供了工业上可行的基础。拓宽了银杏作物的利用价值。
保藏信息
本发明提供的酵母菌A保藏于中国微生物菌种保藏管理委员会普通微生物中心(CGMCC),分类命名为酿酒酵母(Saccharomyces cerevisiae),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,邮政编码100101,该菌株保藏编号为CGMCCNo.28245,保藏日期2023年8月24日。
附图说明
图1发酵处理前的银杏外种皮;
图2发酵处理后的银杏外种皮浆料干燥后样品。
具体实施方式
为使本发明的目的、技术方案和优点更加清楚,下面将对本发明的技术方案进行详细的描述。以下的实施例便于更好地理解本发明,但并不限定本发明。下述实施例中的实验方法,如无特殊说明,均为常规方法。
实施例1
酵母菌A的筛选:从杏果自然发酵液中分离和筛选得到具有异味消除能力的酵母菌A。具体操作是:取北京市黄杏子的自然发酵液10ml,用无菌水稀释,得到稀释液,将稀释液均匀涂布在YPD培养基上,在28℃恒温箱中培养48小时,选取具有典型酵母特征的单菌落,接种到液体培养基中培养,分离出不同的酵母菌株,通过在培养基中添加银杏外种皮选择培养和筛选后,发现其中一株酵母菌株A在接种于银杏外种皮匀浆后,发酵液中银杏外种皮臭味消除明显,而其他菌株对异味消除不明显,将该酵母菌命名为酵母菌 A。酵母菌 A保藏在中国微生物菌种保藏管理委员会普通微生物中心,菌种保藏编号CGMCC NO. 28245。
实施例2
1. 收集银杏果:银杏果实成熟期,收集100kg无明显腐烂或损伤的银杏果,确保获得品质合格原料。
2. 清洗:分拣出的银杏果外种皮放入干净容器中,加入适量清水,充分冲洗果实表面,去除附着在果实上的尘土、泥土和其他污物。
3. 外种皮去除:清洗后的银杏果放入干净的容器中,脱皮机脱除银杏果外种皮。
4. 匀浆处理:约50kg银杏果外种皮置于匀浆机中,加入20L的清水。利用匀浆机将外种皮和水搅拌至均匀的浆状物。持续搅拌15分钟,确保外种皮充分均匀,为后续酵母菌接种提供均质基础。
5. 发酵:调整匀浆的可溶性固形物达15%,pH为3.0-3.3。将活化好的酵母菌A接种于匀浆中,接种量为6%,接种浓度为106cfu/mL,然后25℃下厌氧静置发酵12天。
6. 结果:12天的发酵结束后,取出发酵后的银杏外种皮匀浆,异味消除的效果显著,其中含有5.36wt%乙醇,经过换算,实施例1中。1升水加入2.5kg银杏外种皮,通过上述发酵过程,最终发酵液中含5%的乙醇浓度。
采用高效液相色谱-质谱联用(HPLC-MS)技术,对发酵前后银杏外种皮浆料成分进行分析,银杏外种皮中的银杏酚酸性化合物具有杀菌抑菌、防治病虫害的药理作用,尤其是在农业害虫防治方面具有广阔应用前景。结果如下表1所示:
表1 发酵前后成分对比
请50名志愿受试者,对发酵前后银杏外种皮浆料的气味进行感官评价,结果如下表2所示:
表2 酵母菌A发酵前后感官评价
可见,在受试的50人中,全部能都分辨发酵前后银杏浆料的气味变化;并且50人全部都认为发酵前的银杏浆料具有浓烈的臭味,而发酵后,都认为臭味有不同程度的下降,而且大部分受试者(33/50)认为臭味已经微弱。
图1发酵处理前的银杏外种皮。图2发酵处理后的银杏外种皮浆料干燥后样品。可以看出,发酵后银杏外种皮颜色由棕色变成橙黄色。
发明人还曾经尝试以其他酵母菌,比如毕赤酵母菌和有孢汉逊酵母菌,按照同样的条件对银杏外种皮浆料发酵前后进行50名受试者气味感官评价对比。
结果如下表3和表4。
表3 毕赤酵母菌发酵前后感官评价
表4 孢汉逊酵母菌发酵前后感官评价
可见,其他酵母菌并不能如同本发明筛选得到的酵母菌A那样,经过发酵后消除银杏外种皮的臭味。

Claims (7)

1.一种利用酵母菌消除银杏外种皮臭味的方法,其特征在于,包括以下步骤:将成熟的银杏果实的外种皮和水打浆,制成浆料,将活化的酵母菌A接种于浆料,厌氧发酵;接种前,加入水使调整浆料的可溶性固形物含量为10-15%,加入有机酸使浆料的pH为3-4;所述有机酸选自柠檬酸、苹果酸、酒石酸、草酸中的至少一种;所述厌氧发酵是在20-30℃厌氧条件下发酵10-20天;所述酵母菌A保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏编号为CGMCC NO. 28245。
2.根据权利要求1所述的方法,其特征在于,打浆时,银杏果实的外种皮和水的质量比为3-5:2。
3.根据权利要求1所述的方法,其特征在于,酵母菌活化是将酵母菌A混入3-6wt%的白砂糖水中,酵母菌A浓度为1×103 -5×103 cfu/mL,然后在37-38℃的条件下培养,直到观察到液面有气泡生成,降至常温,得到经过活化的酵母菌A溶液。
4.根据权利要求1所述的方法,其特征在于,酵母菌接种浓度为50-200 cfu/mL。
5.根据权利要求1所述的方法,其特征在于,接种前,加入有机酸使浆料的pH为3-3.3。
6.根据权利要求1所述的方法,其特征在于,包括以下步骤:
(S1)收集无明显腐烂或损伤的成熟银杏果;
(S2)分拣出的银杏果进行清洗,脱除银杏果外种皮;
(S3)银杏果外种皮置于匀浆机中,按照银杏果:清水质量比3-5:2加入清水,将外种皮和水打浆得到均匀的浆状物;
(S4)加入清水调整匀浆的可溶性固形物含量,调节pH为3-4,将活化好的酵母菌A接种于匀浆中,然后20-30℃下厌氧静置发酵10-20天,发酵结束后,取出发酵后的银杏外种皮匀浆;所述酵母菌A保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏编号为CGMCC NO. 28245;
步骤(S4)中,调整匀浆的可溶性固形物含量为10-15%,调节pH至3-3.3。
7.一种用于消除银杏外种皮臭味的酵母菌A,其特征在于,保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏编号为CGMCC NO. 28245。
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