CN114874926A - 类胡萝卜素氧化菌株及其应用和用于烟草加香的浸膏 - Google Patents
类胡萝卜素氧化菌株及其应用和用于烟草加香的浸膏 Download PDFInfo
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- A—HUMAN NECESSITIES
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Abstract
本发明提供了一种类胡萝卜素氧化菌株及其应用和用于烟草加香的浸膏,该菌株于2019年5月27日保藏于中国典型培养物保藏中心,保藏编号为CCTCCNO:M 2019399,保藏名称为酿酒酵母Saccharomyces cerevisiae CFFSH010。其应用于提高烟草提取物熟甜香及提高茶叶体物花果香气。
Description
技术领域
本发明涉及生物转化生香技术领域,特别是类胡萝卜氧化生香领域,具体来说,涉及一种类胡萝卜素氧化菌株及其应用和用于烟草加香的浸膏。
背景技术
类胡萝卜素是异戊二烯类化合物,一般是由两分子的二十个碳原子组成的双香叶基双磷酸尾尾连接而成的,它们具有四十个碳原子的基本结构骨架,不同种类的类胡萝卜素都是在此基础上衍生而来的,但都含有很多的共扼双键。类胡萝卜素的种类很多,按分子结构中是否含有氧原子可分为两大类:不含氧类胡萝卜素(如八氢番茄红素、番茄红素、α-胡萝卜素、β-胡萝卜素等)和含氧的类胡萝卜素(如玉米黄质、叶黄素、花药黄质、虾青素等),含氧的类胡萝卜素又包括双环氧化物和单环氧化物。按碳骨架末端的衍生结构可分为开环式类胡萝卜素(如八氢番茄红素、番茄红素)和环式类胡萝卜素(如α-胡萝卜素、β-胡萝卜素等)。
大量研究表明(茶叶类胡萝卜素合成、代谢调控研究进展,DOI:10.16768/j.issn.1004-874X.2021.05.003.)类胡萝卜素是植物中的重要香气前体,其降解产物对于葡萄酒、茶叶、烟草的香气品质均具有重要的贡献。在大部分植物中,植物内源的类胡萝卜素氧化酶CCD1、CCD4和CCD7具有氧化裂解多种类胡萝卜素生成具有花果香的β-紫罗兰酮、α-紫罗兰酮、香叶基丙酮和二氢海葵内酯等香气物质,这是乌龙茶花果香味形成的重要来源。大马士酮是类胡萝卜素降解形成的另一重要香气物质,常见于加工食品和饮料中,为酒精饮料中的风味成分,常见于葡萄酒中。该香气物质在绿茶和红茶中主要贡献类似水果和蜂蜜的香气特征,浓度很低,但气味感知阈值极低(0.004μg/L),在一些成品茶和茶饮料的风味中发挥重要作用。
自然条件下,葡萄酒、茶叶、烟草都需要经过生物发酵过程来加速生物质香气前体向香气分子转化。除了植物细胞内自身所含有的氧化酶参与以外,微生物的氧化酶体系及自身的代谢也参与植物香原料的代谢过程中,继而形成特有的色、香、味品质。在这过程中,优秀的生香菌株能够明显的加速发酵生香的进程,同时能够提高发酵原料的品质。
发明内容
本发明所要解决的技术问题在于提供一种能够加速类胡萝卜素的氧化,加快香气物质的生成的类胡萝卜素氧化菌株。
其所要解决的技术问题可以通过以下技术方案来实施。
一种类胡萝卜素氧化菌株,其特点为,该菌株于2019年5月27日保藏于中国典型培养物保藏中心,保藏编号为CCTCC NO:M 2019399,保藏名称为酿酒酵母Saccharomycescerevisiae CFFSH010。
本发明所要解决的另一技术问题在于提供一种上述类胡萝卜素氧化菌株在提高烟草提取物熟甜香上的应用,该应用包括采用所述酿酒酵母CFFSH010对烟草提取物进行发酵的步骤。
本发明所要解决的又一技术问题在于提供一种上述类胡萝卜素氧化菌株在提高茶叶体物花果香气上的应用,该应用包括采用所述酿酒酵母CFFSH010对茶叶提取物进行发酵的步骤。
本发明还提供了一种用于烟草加香的浸膏,其利用所述的酿酒酵母CFFSH010对烟草提取物进行发酵获得。
采用上述技术方案的类胡萝卜素氧化菌株及其应用,通过CFFSH010菌株的发酵之后,烟草提取物的酮甜香、熟甜香被大幅的提升了,在极大程度上改善了烟草等品质等级。而CFFSH010菌株的发酵应用于茶叶提取物上,则极大程度的使得原有的茶香中的果甜香及花香被极大的增强,非常适合于红茶和普洱茶等发酵或半发酵型茶茶香产品的品质提升。其改善烟叶提取物和茶叶提取物的香气品质的原因,可能在与菌株CFFSH010菌株具有良好的类胡萝卜素降解代谢途径及相关酶,其能够极大的增加类胡萝卜素产物类香气成分含量及丰度。
附图说明
图1为以CFFSH010为发酵菌株发酵后的云烟浸膏的GC-MS谱图;
图2为未经发酵的云烟浸膏的GC-MS谱图
图3为不同发酵时间获得的云烟浸膏的相对大马酮含量的变化图;
图4为不同发酵温度获得的云烟浸膏的相对大马酮含量。
具体实施方式
本发明提供一种类胡萝卜素氧化菌株,其能够加速类胡萝卜素的氧化,加快香气物质的生成。本发明所提供的菌株为保藏编号为CCTCC NO:M 2019399的酿酒酵母CFFSH010。
本发明还提供一种提高烟草提取物熟甜香的方法,该方法使用酿酒酵母CFFSH010对于烟草提取物进行发酵。
本发明同时提供一种提高茶叶体物花果香气的方法,该方法使用酿酒酵母CFFSH010对于茶叶提取物进行发酵。
以下为本发明更具体的实施例。
实施例1:对烟草提取物的发酵
1)种子液的制备:
配置YPD液体培养基(1%Yeast Extract(酵母膏),2%Peptone(蛋白胨),2%Dextrose葡萄糖),115℃灭菌20min,培养基降至室温后,分别接入1%w/v)筛选的菌株CFFSH010及其他对照菌株(菌株清单见表1),30℃培养12h。
表1:接种菌株种类及公开来源
2)烟草浸膏发酵物的制备:
云烟浸膏(深圳波顿香料有限公司提供),110℃灭菌15min,降至室温,按1%(w/w)接种量进行接种,搅拌均匀后,于40℃发酵7天。
3)发酵后处理
采用GC/MS分析。发酵结束后,取烟草浸膏固相微萃取后,进样分析,以类胡萝卜素氧化降解产物大马酮为目标分析产物。以CFFSH010为发酵菌株发酵后的谱图如图1所示,其中β-大马酮的峰高和峰面积相比于发酵前的原始云烟浸膏(GC-MS图谱如图2)有大幅提升,其中保留时间(RT)57.7min的β-大马酮(Damascone)峰面积提高到了2143%,与此同时另一类胡萝卜的降解产物烟酮(RT 48.1min)其产物的相对峰面积也有较大提升,提升幅度达205%。
其他各对照菌株发酵的发酵结果如表2所示,大部分测试的其他酵母仅少量增加,其中最高的CICC 33328增加2倍多(增加至原含量的325%),新良高活性干酵母粉甚至在大马酮含量上有所减少。而自筛的特异性菌株使得β-大马酮含量大幅度提升了21.4倍。
表2:发酵之后β-大马酮的相对含量
4)感官评价:
发酵结束后,将烟草浸膏用饮用水稀释100倍后,以喷雾器均匀喷至空白烟丝上(每100g烟丝喷1g稀释液),之后将烟丝置于恒温恒湿箱中平衡烟丝水分至烟丝水含量12.5-13.5%w/w,之后将烟丝卷制成烟后,按照国标GB 5606.4感官技术要求的评价标准进行评吸,选10名资质评委,综合汇总结果见表3。
感官评价结果表明,以CFFSH010发酵后的云烟浸膏显著性的增加了云烟浸膏的品质,评吸评委给出的感官评定综合意见为:显著的增加了香气量,香气量充足,香气较透发,烟气较细腻、柔和,降低了刺激,刺激较小,余味较干净,回甜较好,同时显著增加了云烟的成熟感和清甜香特征,降低了青杂气息。
表3:发酵后的云烟浸膏的功能感官评价
实施例2:对红茶提取物的发酵
1)种子液的制备:
配置YPD液体培养基(1%Yeast Extract(酵母膏),2%Peptone(蛋白胨),2%Dextrose葡萄糖),115℃灭菌20min,培养基降至室温后,分别接入1%v/v)筛选的菌株CFFSH010及其他对照菌株(菌株清单见表1),30℃培养12h。
2)红茶比例提取物的制备:
红茶棕黄色精细粉末(深圳波顿香料有限公司提供),110℃灭菌15min,降至室温,按1%(w/w)接种量将种子液均匀喷撒在棕黄色精细粉末上,于恒温恒湿箱30℃,RH80%,发酵7天。发酵完成之后,以85℃水进行冲泡。茶水中茶粉质量含量为10%(即10g茶每100g茶水),冷却至室温之后进行香气评价与β-大马酮含量测定(GC-MS法)。
发酵后的β-大马酮的相对含量如表4所示,结果表明CFFSH010能够显著增加红茶提取物β-大马酮的含量。通过评香人员的香气评价表面,通过CFFSH010的发酵的红茶提取物其花香和果香的香气量和香气质都有了明显的提升,同时红茶提取物的成熟感及甜感、余味均有显著的提高。
表4:发酵之后β-大马酮的相对含量
实施例3:发酵条件优化
配置100mL YPD液体培养基(1%Yeast Extract(酵母膏),2%Peptone(蛋白胨),2%Dextrose葡萄糖),115℃灭菌20min,培养基降至室温后,以接种接入1环菌株CFFSH010的菌苔,于30℃培养,220rpm振荡培养18h,作为种子液。
之后,按照不同优化条件进行云烟浸膏的发酵条件优化。
(a)接种量的优化
云烟浸膏(深圳波顿香料有限公司提供),110℃灭菌15min,降温至室温,按照不同的接种量对云烟浸膏进行接种,之后将置于30℃下恒温发酵7天。按照实施例1所述方式对云烟浸膏中β-大马酮相对含量与感官评价。结果如表5所示。接种量在1%至10%范围内对发酵7天后β-大马酮的含量并无显著性影响,同时,所获的的发酵后浸膏其吃味比较接近,没有显著的风格差异,但是当菌株的接种量超过7%以后,会略微增加烟气中的杂气。
表5:不同接种量对CFFSH010发酵云烟浸膏的影响
(b)培养时间的优化
将云烟浸膏(深圳波顿香料有限公司提供),110℃灭菌15min,降温至室温,按照1%w/w接种量对云烟浸膏进行接种,之后将置于30℃下恒温发酵0至30天,定期取样并按照实施例1所述的方法检测发酵过程中β-大马酮的相对含量。按照实施例1所述方式对云烟浸膏中β-大马酮相对含量。以未接种任何菌株的云烟浸膏(深圳波顿香料有限公司提供),110℃灭菌15min,降温至室温,之后于30℃下恒温放置0-30天作为阴性对照。结果如图3所示。结果表明采用CFFSH010菌株发酵后,其在发酵第7天后,β-大马酮就可达到较高水平。
(c)培养温度的优化
将云烟浸膏(深圳波顿香料有限公司提供),110℃灭菌15min,降温至室温,按照1%w/w接种量对云烟浸膏进行接种,之后将置于不同的优化温度下下发酵7天,之后按照实施例1所述的方法,检测发酵结束后云烟浸膏中β-大马酮的相对含量,结果如图4所示。结果表明,菌株CFFSH010具有较为宽泛的温度适宜范围,在20℃-45℃范围或者自然室温(不控温)下内都能适宜生存,并且能够很好的发挥其增加β-大马酮含量的生物生香的生理学功能。但在50℃以上高温环境,其生理生化活性可能受到影响,生物生香功能明显降低。
Claims (6)
1.一种类胡萝卜素氧化菌株,其特征在于,该菌株于2019年5月27日保藏于中国典型培养物保藏中心,保藏编号为CCTCC NO:M 2019399,保藏名称为酿酒酵母Saccharomycescerevisiae CFFSH010。
2.权利要求1所述类胡萝卜素氧化菌株在提高烟草提取物熟甜香上的应用,其特征在于,包括采用所述酿酒酵母CFFSH010对烟草提取物进行发酵的步骤。
3.权利要求1所述类胡萝卜素氧化菌株在提高茶叶体物花果香气上的应用,其特征在于,包括采用所述酿酒酵母CFFSH010对茶叶提取物进行发酵的步骤。
4.根据权利要求1或2所述的应用,其特征在于所述的酿酒酵母CFFSH010的发酵温度为20至45℃。
5.根据权利要求1或2所述的应用,其特征在于所述的酿酒酵母CFFSH010的发酵时间为至少7天。
6.一种利用权利要求1所述类胡萝卜素氧化菌株制备的用于烟草加香的浸膏,其特征在于,利用所述的酿酒酵母CFFSH010对烟草提取物进行发酵获得。
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