CN105779130A - 一种发酵制备万寿菊挥发油的方法及其应用 - Google Patents
一种发酵制备万寿菊挥发油的方法及其应用 Download PDFInfo
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- CN105779130A CN105779130A CN201610278233.7A CN201610278233A CN105779130A CN 105779130 A CN105779130 A CN 105779130A CN 201610278233 A CN201610278233 A CN 201610278233A CN 105779130 A CN105779130 A CN 105779130A
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- marigold
- tagetis erectae
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Abstract
本发明提供一种β‑葡萄糖苷酶发酵制备万寿菊挥发油的方法，包括以下步骤：(1)将万寿菊花与水按质量比1:10～40混合，乙酸调pH至4～6，在45～55℃条件下平衡30‑60分钟，加入占万寿菊花质量比0.1～0.6％的β‑葡萄糖苷酶，混合均匀并不停搅拌、45～55℃温育12～48小时；(2)水蒸气蒸馏5～10小时提取挥发油，所得黄绿色油状物为酶发酵提取万寿菊挥发油。本发明经β‑葡萄糖苷酶发酵万寿菊花提取挥发油，其特征香气集中，带来刺激和杂气的挥发性物质消失或减少；在再造烟叶中应用改善了未经酶发酵的万寿菊挥发油在再造烟叶中与烟气不协调、粗糙、刺激性大、杂气重、掩盖烟香的缺陷。
Description
技术领域
本发明涉及烟用香料技术领域，具体是一种发酵制备万寿菊挥发油的方法及其在再造烟叶加香中的应用。
背景技术
再造烟叶，主要由烟末、碎片、烟梗或低次烟叶加入胶粘剂和其他添加剂等组成，存在杂气重、刺激性大和口感不适等吸味缺陷，影响了在卷烟产品中的使用效果和添加量。通常采用添加剂的办法来进行改善，目前应用较多的添加剂是烟草、天然植物类提取物或挥发油等，这些提取物一般通过水、醇提取、水蒸气蒸馏或分子蒸馏得到，添加后对卷烟质量的改善程度有限，仍有刺激性大、杂气多、香气不足等缺陷。
万寿菊(TagetserectaL)是菊科万寿菊属一年生草本植物，橙色品种万寿菊干花朵类胡萝卜素含量为0.6％～2.5％，其中叶黄素酯约占总类胡萝卜素88％～92％。万寿菊精油，用以调制拟香型优雅的香水、花露水和生产芳香医疗型化妆品。目前对于万寿菊的研究多集中于叶黄素方面，对于精油的化学成分也有一定的研究，万寿菊精油具有抗氧化、驱蚊、抑菌等生物活性。据报道万寿菊叶的挥发油的水蒸气蒸馏法的提取率有0.6-0.9％。
本发明人在研究中发现：天然植物性香料中部分发香物质的发香基团往往容易与植物细胞中的其他成分如低分子的糖类等以化学键合的形式相结合，从而抑制了香味的散发并难以被提取，在香料科学领域内将该类物质称为“香味前驱体”。这些物质本身并无香味且不挥发，但若选择合适的酶进行水解后，可释放出浓郁的具有天然特征香味的物质。β-葡萄糖苷酶能够水解结合于末端非还原性的β-D-葡萄糖苷键，同时释放出β-D-葡萄糖和相应的配基。此外，β-葡萄糖苷酶能将果、蔬、茶中的风味前体物质水解为具有浓郁天然风味的香气物质，还能降解或协同降解纤维素。
现有技术中，以万寿菊花为原料，β-葡萄糖苷酶发酵制备万寿菊挥发油，提高提取率和香气品质，并在再造烟叶中加香应用，改善烟气的香气质、口感，降低刺激减少杂气，尚未见报道。
发明内容
本发明针对现有技术的不足，提供一种β-葡萄糖苷酶发酵制备万寿菊挥发油，使万寿菊花的香味前体物经酶的作用产生降解、分解、转化生成丰富的、小分子的香气物质，而增加刺激感和杂气的物质含量减少甚至消失，并且提高万寿菊挥发油的提取率；以及该发酵万寿菊挥发油在在再造烟叶中应用，丰富香气、提调烟香、口感，降低刺激、减少杂气等。
本发明提供一种β-葡萄糖苷酶发酵制备万寿菊挥发油的方法，包括以下步骤：
(1)将万寿菊花与水按质量比1:10～40混合，乙酸调pH至4～6，在45～55℃条件下平衡30-60分钟，加入占万寿菊花质量比0.1～0.6％的β-葡萄糖苷酶，混合均匀并不停搅拌、45～55℃温育12～48小时；
(2)水蒸气蒸馏5～10小时提取挥发油，所得黄绿色油状物为酶发酵提取万寿菊挥发油。
本发明提供一种上述方法制备的万寿菊挥发油在再造烟叶中的应用，所述万寿菊挥发油占再造烟叶的质量比为0.01～0.03％。
本发明对β-葡萄糖苷酶发酵万寿菊制备的挥发油与未经酶发酵的万寿菊挥发油对照(以下简称对照)进行挥发性致香成分GC/MS对比分析。
(1)挥发油处理：将挥发油用二氯甲烷萃取、稀释100倍，取1～2mL用于进行GC/MS分析。
(2)GC/MS分析：HP-5MS(50m×0.25mm×0.25μm)毛细管柱；进样温度：250℃；分流比20:1；载气：He，1mL/min；升温程序：从50℃(1min)以5℃/min升到250℃(5min)；传输线温度：250℃；离子源：EI源；电子能量：70eV；扫描范围：50～550amu；使用WILEY和MINILAB谱库进行检索。
(3)挥发性成分对比分析
GC/MS分析结果(表1)表明，
①未经酶发酵处理的万寿菊挥发油中共分离鉴定出98种挥发性香气成分，主要包括烯类、萜烯类、环烃族化合物、萜类、醇类、酮类、醛类、烷烃类，还有呋喃、吡嗪、噻吩等杂环类化合物，占74.61％，其中柠檬烯(5.02％)、4—蒈烯(5.5％)、斯巴醇(8.12％)、石竹烯氧化物(4.13％)、肉豆蔻酸(6.78％)、棕榈酸(6.10％)等，占35.65％；
②经β-葡萄糖苷酶发酵制备的万寿菊挥发油中共分离鉴定出127种挥发性香气成分，相对百分含量占95.04％，其中柠檬烯(3.61％)、4-蒈烯(4.62％)、β-石竹烯(16.82％)、斯巴醇(11.03％)、石竹烯氧化物(2.25％)、肉豆蔻酸(5.90％)、棕榈酸(5.98％)等，占50.21％；经酶促发酵生香后，万寿菊挥发油中的香气成分更丰富，转化出了新的香气成分β-石竹烯，并且与其他6种香气成分占了总挥发性成分的50％以上，集中了优势香气，构成了酶促生香万寿菊挥发油的特征香气，提升了香气品质；
③经β-葡萄糖苷酶发酵制备的万寿菊挥发油中新出现了70种香气成分，主要有β-石竹烯(16.818％)、β-水芹烯、柠檬醛、4,7-二甲基苯并呋喃、优葛缕酮、E-香芹醇、香芹酮、百里香酚、(+)-α-长叶蒎烯、α-荜澄茄油烯、(+)-香橙烯、2,3-二氢-2-甲氧苯并呋喃、双环大根香叶烯、二氢-(-)新丁香三环烯-(I)、o-薄荷-8-烯、喇叭茶醇、桃醛、金合欢基丙酮、丁位壬内酯、亚麻酸甲酯、硬脂酸等。其中，β-石竹烯(16.818％)是一种双环倍半萜类化合物，具有辛香、木香、柑橘香、樟脑香，温和的丁香香气，有平喘、镇咳、祛痰作用；桉油烯醇(11.031％)在多种花香油中天然存在；β-水芹烯具柑橘、胡椒的清凉香气；柠檬醛有柠檬香气；香芹酮是一种重要的香料，它广泛应用于食品工业和化学工业中，特别是牙膏，硬糖，口香糖和各种饮料中，因此(-)-香芹酮的合成具有重要的经济价值；百里香酚是一个单萜酚，有有百里香油似的辛香和草香香气；(+)-α-长叶蒎烯、α-荜澄茄油烯、(+)-香橙烯、斯巴醇均为萜烯类化合物，具有香气；桃醛有强烈的桃子和杏仁样香气，是一种重要的内酯香料；金合欢基丙酮具有淡的青甜香气，增加青甜香和烤烟香；丁位壬内酯具有甜的、温暖的坚果、牛奶、奶油、椰子样气味；亚麻酸甲酯为不饱和脂肪酸，是构成卷烟香味的重要物质，柔和烟气，减轻刺激；硬脂酸赋予烟草脂肪和脂腊样香气；
④经β-葡萄糖苷酶发酵制备的万寿菊挥发油中龙脑、薄荷醇、苯乙酸甲酯、三甲基苯甲醇、臧红花醛、薄荷酮环氧化物、乙酸香叶酯、月桂酸甲酯、月桂酸、肉豆蔻酸甲酯、十五碳酸甲酯、植酮、棕榈酸甲酯、苯并(a)吩嗪、亚油酸甲酯、硬脂酸甲酯等成分的含量有不同程度地提高。其中龙脑香气清凉尖刺，微带药香、胡椒香，香气飘；薄荷醇具有薄荷香气并有清凉的作用；苯乙酸甲酯天然存在于可可、咖啡及草莓中，有弱的蜂蜜及麝香样香气，略有甜味；臧红花醛对多种癌细胞均有明显抑制作用，具有木香、辛香、药香、粉香；乙酸香叶酯有玫瑰和熏衣草香气；月桂酸甲酯具有类似酒香及花香；月桂酸微有月桂油香味；肉豆蔻酸甲酯具似蜂蜜和鸢尾样的香气；亚油酸甲酯为不饱和脂肪酸，是构成卷烟香味的重要物质，柔和烟气，减轻刺激；
⑤酶发酵后，万寿菊挥发油中的一些成分消失或含量降低：柠檬烯、2,3-二氢-2-甲基苯并呋喃、右旋樟脑、(-)-4-萜品醇、胡薄荷酮、2-乙酰基-3,5-二甲基吡嗪、枯茗醛、右旋香芹酮、溴乙酸苯乙酯、(1S)-(+)-3-蒈烯、香芹酚、1,2-二氢-1,1,6-三甲基萘、(-)-α-古芸烯、α-紫罗酮、4-(2,6,6-三甲基-1,3-环己二烯)-3-丁烯-2-酮、巴伦西亚橘烯、β-瑟林烯、白菖烯环氧化物、石竹烯乙醇、石竹烯氧化物、3,7,7-三甲基-双环[4.1.0]庚-2-烯等杂环烯烃和杂环化合物，可能是在β-葡萄糖苷酶的作用下，有些成分发生了结构变化形成了同分异构体，而杂环烯烃、杂环化合物等大分子物质断裂分解、降解，产生更多的小分子香气物质或者转化成别的挥发性香气成分甚至消失，可能有利于卷烟品质的提高。
总之，从挥发、半挥发性成分的变化看，经β-葡萄糖苷酶发酵的万寿菊挥发油在再造烟叶中的加香效果可能比对照好。
表1β-葡萄糖苷酶发酵提取万寿菊挥发油与其对照的挥发性成分GC/MS对比
表1中符号“/”表示未检出；内标为丙酸苯乙酯。
(4)在再造烟叶中加香的感官评价分析
经过再造烟叶加香评吸试验(表2)表明，经β-葡萄糖苷酶发酵制备的万寿菊挥发油在再造烟叶中的加香效果明显好于未酶处理的万寿菊挥发油(简称对照)，香气特征表现得更为饱满、突出，香气丰富、香气质提高，烟气更加柔和细腻，刺激性、杂气明显降低，带有清凉回甜感。适宜加香量为0.01％。
表2酶发酵万寿菊挥发油及其对照样品在再造烟叶中的加香评吸结果
本发明的优点是：经β-葡萄糖苷酶发酵万寿菊花提取挥发油，其挥发油的香气成分变化大，香气更丰富、香气质提高，特征香气集中，带来刺激和杂气的挥发性物质消失或减少，大分子物质经酶促生香降解、分解、转化成小分子香气物质甚至消失，有的转化为同分异构体；在再造烟叶中应用，具有协调烟气、增加清甜清凉感、降低刺激、减少杂气、改善吸味、增强喉部舒适性的优点，能与其它烟用香精香料混合调配使用，改善了未经酶解的万寿菊挥发油在再造烟叶中与烟气不协调、粗糙、刺激性大、杂气重、掩盖烟香的缺陷。该方法所用原料来源广泛，仪器简单，易于操作和实现产业化，成本低，绿色环保，旨在为酶促生香开发天然烟用香料的应用提供参考。
具体实施方式
下面列举一部分实施例对本发明的有关技术问题作进一步详细的描述，有必要在此指出以下具体实施例只是对本发明的进一步说明，不代表对本发明保护范围的限制。其他人根据本发明做出的一些非本质的修改和调整，仍属于本发明的保护范围。
实施例1
(1)将万寿菊粗粉与水按质量比1:10混合，乙酸调pH至4.5，在45℃条件下平衡半小时，加入占万寿菊粗粉质量比0.1％的β-葡萄糖苷酶混合均匀，并不停搅拌、45℃温育12小时；
(2)水蒸气蒸馏5小时提取挥发油，所得黄绿色油状物为酶发酵万寿菊挥发油；
将上述万寿菊酶发酵制备挥发油在再造烟叶中加香，占再造烟叶的质量比为0.01％。
感官评吸效果：与烟气协调性较好，有清甜清凉感，烟气柔和、刺激降低、杂气减少、余味干净，香气质提高。
实施例2
(1)将万寿菊粗粉与水按质量比1:20混合，乙酸调pH至5.0，在50℃条件下平衡40分钟，加入占万寿菊粗粉质量比0.3％的β-葡萄糖苷酶混合均匀，并不停搅拌、50℃温育18小时；
(2)水蒸气蒸馏6小时提取挥发油，所得黄绿色油状物为酶促生香万寿菊挥发油；
将上述经β-葡萄糖苷酶发酵制备的万寿菊挥发油在再造烟叶中加香，占再造烟叶的质量比为0.01％。
感官评吸效果：与烟气协调性好，香气更丰富，香气质提高，喉部舒适，烟气柔和细腻有清凉回甜感，刺激明显降低，杂气减少，余味干净。
实施例3
(1)将万寿菊粗粉与水按质量比1:40混合，乙酸调pH至6，在55℃条件下平衡45分钟，加入占万寿菊粗粉质量比0.5％的β-葡萄糖苷酶混合均匀，并不停搅拌、55℃温育32小时；
(2)水蒸气蒸馏10小时提取挥发油，所得黄绿色油状物为酶促生香万寿菊挥发油；
将上述经β-葡萄糖苷酶发酵制备的万寿菊挥发油在再造烟叶中加香，占再造烟叶的质量比为0.01％。
感官评吸效果：与烟气协调性好，喉部有清凉甜感，舒适，烟气柔和、刺激降低、杂气减少、余味干净，香气质提高。
以上所述，仅为本发明的具体实施方式，但本发明的保护范围并不局限于此，任何属于本技术领域的技术人员在本发明揭露的技术范围内，可轻易想到的变化或替换，都应涵盖在本发明的保护范围之内。因此，本发明的保护范围应该以权利要求的保护范围为准。
Claims (2)
1.一种β-葡萄糖苷酶发酵制备万寿菊挥发油的方法，其特征在于包括以下步骤：
(1)将万寿菊花与水按质量比1:10～40混合，乙酸调pH至4～6，在45～55℃条件下平衡30-60分钟，加入占万寿菊花质量比0.1～0.6％的β-葡萄糖苷酶，混合均匀并不停搅拌、45～55℃温育12～48小时；
(2)水蒸气蒸馏5～10小时提取挥发油，所得黄绿色油状物为酶发酵提取万寿菊挥发油。
2.一种如权利要求1所述方法制备的万寿菊挥发油在再造烟叶中的应用，所述万寿菊挥发油占再造烟叶的质量比为0.01～0.03％。
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| CN106281699A (zh) * | 2016-08-16 | 2017-01-04 | 重庆雅香美源生态农业科技有限公司 | 一种万寿菊精油的提取及处理方法 |
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| CN106281699A (zh) * | 2016-08-16 | 2017-01-04 | 重庆雅香美源生态农业科技有限公司 | 一种万寿菊精油的提取及处理方法 |
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| CN106978258A (zh) * | 2017-05-24 | 2017-07-25 | 湖北中烟工业有限责任公司 | 酶促增香提高香气品质的杭白菊挥发油的制备方法及应用 |
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