CN110432481A - 低糖蔬果酵素液的制作方法 - Google Patents
低糖蔬果酵素液的制作方法 Download PDFInfo
- Publication number
- CN110432481A CN110432481A CN201910359364.1A CN201910359364A CN110432481A CN 110432481 A CN110432481 A CN 110432481A CN 201910359364 A CN201910359364 A CN 201910359364A CN 110432481 A CN110432481 A CN 110432481A
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- Prior art keywords
- tunning
- enzyme liquid
- vegetables
- fruits
- production method
- Prior art date
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Abstract
本发明所揭低糖蔬果酵素液的制作方法为将至少一蔬果至少一蔬果作为发酵原料，并依序进行至少三次发酵程序，以获得一酵素液，其中：该第一次发酵程序的菌元为酵母菌；该第二次发酵程序的菌元为醋酸菌；该第三次发酵程序的菌元为乳酸菌；并且，该酵素液的糖份低于5％，并且在一预定条件下，该酵素液的糖份低于2.5％。
Description
技术领域
本发明有关于一种发酵技术，特别指一种低糖蔬果酵素液的制作方法。
背景技术
微生物发酵技术为通过特定微生物与原料共同进行培养的技术。过去仅将微生物发酵技术从应用制造各类发酵食品，如乳酪、优格、酱油、啤酒、泡菜，而随着生物科技及医学研究的进步，许多研究指出经微生物进行代谢或分解的产物对于生物体有许多益处，因此，微生物发酵技术开始被应用产制各类型的酵素产品。
虽然酵素产品被认为对于人体具有益处，例如促进消化、分解脂肪等，但，事实上，目前市面上的酵素产品利用蔬果作为基材进行发酵，因为蔬菜水果本身带有甜度，尤其是水果的甜度更高，使得目前市面上所贩售的蔬果酵素具有高糖份的特性，意即蔬果酵素含有高于50％的糖份。而高糖份的酵素产品不仅无法提供给罹患代谢症候群或是糖尿病的患者使用，更会提高罹患脂肪肝、癌症、肥胖等风险。
发明内容
本发明的主要目的在于提供一种低糖蔬果酵素液的制作方法，其通过至少三道发酵程序进行蔬果原料的发酵培养，以达到降低终产物糖度的功效。
本发明的另一目的在于提供一种低糖蔬果酵素液的制作方法，其能够大量产制低糖高纤的酵素液，以能提供产制酵素相关产品使用。
于是，为能达成上述目的，本发明所揭低糖蔬果酵素液的制作方法为将至少一蔬果作为发酵原料，并依序进行至少三次发酵程序，以获得一酵素液，其中：该第一次发酵程序的菌元为酵母菌；该第二次发酵程序的菌元为醋酸菌；该第三次发酵程序的菌元为乳酸菌；并且，该酵素液的糖份低于5％。
更进一步来说，本发明所揭低糖蔬果酵素液的制作方法，其包含下列步骤：
步骤a：取一培养基，具有至少一蔬果原料，其中，该培养基的糖度为15～25度。
而可使用下列方式来调整该培养基糖度：添加如砂糖、黑糖、糖蜜等增甜剂于蔬果原料中进行均匀混合。而可以HPLC检测该培养基内的糖份含量，并再计算出增甜剂的添加量。
步骤b：取一活化的酵母菌，如Saccharomyces cerevisiae，并以一预定接种量将该酵母菌接种于该培养基上，进行无氧发酵，当发酵至发酵产物的残糖含量低于2％及酒精含量为4.5～5.5％时，得到一第一发酵产物。
其中，该预定接种量为0.01～1％w/v，如0.01、0.02、0.03、0.05、0.08、0.1、0.2、0.3、0.5、0.6、0.8、0.9、1.0％w/v，其中，又以接种量为0.1％w/v为佳。
步骤c：取一醋酸菌，如Acetobacter aceti，并以1～10％w/v的接种量将该醋酸菌接种于该第一发酵产物上，进行有氧培养，发酵温度为35～40℃，当发酵至发酵产物的醋酸浓度为1.8％～2.5％时，得到一第二发酵产物。
其中，该预定接种量为1～10％w/v，如1、2、2.5、3、4、5、5.5、6、6.8、7、8、8.5、9、10％w/v；又以接种量为5％w/v者为佳。
步骤d：一芽孢乳酸菌，并以5～20％w/v接种量将该乳酸菌接种于该第二发酵产物上，进行通气培养，当发酵至发酵产物的乳酸含量大于600ppm时，得到一第三发酵产物，并该第三发酵产物的糖份低于5％。
该预定接种量为5～20％w/v，如5、6、6.5、7、8、9、10、11、11.5、12、12.3、13、14、14.2、15、15.8、16、17、17.6、18、19、20％w/v；又以接种量为5％w/v者为佳。
步骤e：过滤该第三发酵产物而得到该酵素液。
在本发明的一实施例中，该当该发酵原料中的至少一蔬果选自由柑橘、木瓜、凤梨、香蕉、奇异果、来姆、葡萄所组成的群，并该蔬果的重量大于等于该发酵原料重量的50％时，该酵素液的糖份低于2.5％。
在本发明的另一实施例中，该第二发酵产物的糖度低于第一发酵产物的糖度至少约10度。
在本发明的另一实施例中，该第一发酵产物的酒精度较未发酵前上升至少3％，其中，又以大于4％较佳。
在本发明的一实施例中，为能使该酵素液具有较为浓烈的酸味，该第二发酵产物的醋酸浓度大于2％。
在本发明的另一实施例中，为能使该酵素液的口感较佳，该第二发酵产物的酒精度小于1％。
在本发明的又一实施例中，该第三发酵产物的酒精度小于1％。
在本发明的次一实施例中，该第三发酵产物的醋酸浓度大于2％。
而为能使菌元能够均匀地进行发酵培养，各该发酵程序为在该发酵程序在一搅动环境下进行，其中，该搅动环境可通过人为或机器来达到。
本发明的有益效果在于：
本发明提供一种低糖蔬果酵素液的制作方法，其通过至少三道发酵程序进行蔬果原料的发酵培养，以达到降低终产物糖度的功效。通过本发明所揭低糖蔬果酵素液的制作方法能将不同甜度的蔬果制备为糖份低于5％的酵素液，并且随着其使用的原料不同，能使酵素液具有不同酸度及酒精度，而给予消费者不同程度的酸味及酒味。
具体实施方式
本发明揭露一种低糖蔬果酵素液的制作方法，其为将含有至少一蔬果原料的培养基通过多重菌种的多道发酵程序进行发酵培养后，而能够得到一糖份低于5％且具不同风味的酵素液。换言之，通过本发明所揭低糖蔬果酵素液的制作方法能将不同甜度的蔬果制备为糖份低于5％的酵素液，并且随着其使用的原料不同，能使酵素液具有不同酸度及酒精度，而给予消费者不同程度的酸味及酒味。
更进一步来说，本发明所揭低糖蔬果酵素液的制作方法至少包含三个发酵程序，其中：
第一发酵程序为酵母发酵程序，所使用的菌元为酵母菌，如：Saccharomycescerevisiae。
第二发酵程序为醋酸发酵程序，所使用的菌元为醋酸菌，如：Acetobacter aceti。
第三发酵程序为乳酸发酵程序，所使用的菌元为乳酸菌，如芽孢乳酸菌。
而依序进行上述三个发酵程序后，再过滤分离残渣，即能获得该酵素液。
本发明所揭以HPLC分析糖度、酒精度及酸度的方法，其为将待测样品中的有机酸、酒精、单双糖或糖醇经过萃取或是过滤后，以HPLC管柱分离，用折射率RI检测器(Refractive Index Detector)与UV-VIS侦测器，并用外标法进行定量，其中，有机酸包含有乳酸、醋酸、柠檬酸、酒石酸、苹果酸；单双糖及糖醇包含有葡萄糖、果糖、蔗糖、麦芽糖、山梨醇等。
举例来说，本发明及其实例中所使用的HPLC检测方法如下：
先将如乳酸、冰醋酸等有机酸或酒精的标准品取2.5g加水定容于50ml，浓度为50mg/ml，再以HPLC进行分析，以确认标准品的滞留时间。将酵素液5ml，加水定容至10ml，以4000rmp在25℃下进行离心5分钟，取上清液，过滤后，以HPLC进行分析，再以外标法得到各酵素液中有机酸、酒精的含量。
而有机酸及酒精的HPLC分析条件为：管柱为IC Sep ICE-Coregel 87H3，移动相为25mM硫酸，流速为0.5ml/min，温度为40℃，注入体积为20μl，分析时间为40分钟；而用于检测醇类的仪器为RI检测器(Refractive Index Detector)，检测有机酸的仪器为UV-VIS侦测器或RI检测器。
单双糖的HPLC分析条件为：管柱为YMC-Pack polyamine II，250x4.6mm I.D.，Particle size:5μm，移动相为乙腈/水(750ml/250ml)，流速为0.7ml/min，温度为25℃，注入体积为20μl，分析时间为60分钟，检测仪器为RI检测器。
以下，为能更进一步说明本发明，将兹举一实施例进行详细说明如后。
本发明的一实施例所揭低糖蔬果酵素液的制作方法，其包含下列步骤：
步骤101：制备培养基
取一选定的蔬果，清洗后，进行裁切，再将之破碎打泥，进行酵素水解，例如果胶分解酵素、纤维分解酵素或是本领域技术人员所知的水解酵素，而后再以添加砂糖的方式调整其糖度，以获得一培养基，其中，该培养基的糖度为20度，单双醣加总含量为14％。
更进一步来说，以HPLC进行原料水解后果泥的单双糖含量的确认，再以下列公式计算出添加砂糖的量：
补足砂糖量＝[原料总重量×(0.14-原料单双糖含量(％))]÷0.86
步骤102：酵母发酵程序
取菌种为Saccharomyces cerevisiae的酵母菌，接种量为0.1％w/v(每毫升有效菌量需达109CFU)，例如500L蔬果汁需500mL酵母菌液，此阶段进行无氧发酵，而在搅拌速率为50rpm、温度为28℃的环境下进行发酵，在发酵第5日，以HPLC监控发酵产物的残糖含量及酒精含量，当残糖含量小于2％，酒精含量为4.5～5.5％，结束酵母发酵程序，得到一第一发酵产物，并进行下一阶段的醋酸发酵程序。
不同蔬果原料进行酵母发酵后得到的第一发酵产物，分别以HPLC进行检测后的结果如下表一所示。
表一：以HPLC检测各该第一发酵产物的酸碱值、总单双糖量、酒精含量的结果
步骤103：醋酸发酵程序
将该第一发酵产物进行醋酸发酵程序。详言之，取菌种为Acetobacter aceti的醋酸菌，接种量为5％w/v(每毫升有效菌量需达108CFU)，此阶段进行有氧发酵，其中，通气量约为0.5VVM，搅拌速率约为200rpm，发酵温度须控制在37℃上下，且不得超过40℃，维持48小时，当发酵至醋酸浓度为1.8％～2.5％时，结束醋酸发酵程序，获得一第二发酵产物，意即该第二发酵产物的醋酸浓度为1.8％～2.5％。
以HPLC检测各该第二发酵产物，结果如下表二所示。
表二：以HPLC检测各该第二发酵产物的酸碱值、总单双糖量、酒精含量、醋酸含量的结果
| 原料 | pH | 总单双糖 | 酒精含量 | 醋酸含量 |
| 柑橘 | 3.03±0.10 | 0.21％±0.09％ | 0.16％±0.05％ | 2.13％±0.05％ |
| 木瓜 | 3.27±0.18 | 0.27％±0.12％ | 0.33％±0.04％ | 2.19％±0.05％ |
| 凤梨 | 3.13±0.11 | 0.16％±0.07％ | 0.21％±0.03％ | 2.01％±0.05％ |
| 香蕉 | 3.33±0.19 | 0.19％±0.08％ | 0.27％±0.04％ | 2.27％±0.06％ |
| 青梅 | 2.99±0.06 | 0.15％±0.06％ | 0.09％±0.03％ | 2.25％±0.10％ |
| 诺丽果 | 3.51±0.28 | 0.47％±0.08％ | 0.05％±0.02％ | 2.25％±0.06％ |
| 香檬皮 | 2.87±0.19 | 0.29％±0.16％ | 0.11％±0.02％ | 2.03％±0.08％ |
| 葡萄柚 | 3.58±0.26 | 0.09％±0.03％ | 0.25％±0.04％ | 2.36％±0.08％ |
| 奇异果 | 3.36±0.20 | 0.10％±0.02％ | 0.46％±0.04％ | 2.38％±0.05％ |
| 莱姆 | 2.88±0.16 | 0.35％±0.09％ | 0.23％±0.01％ | 2.17％±0.09％ |
| 葡萄 | 3.33±0.26 | 0.47％±0.17％ | 0.28％±0.06％ | 2.08％±0.05％ |
步骤104：乳酸发酵程序
将该第二发酵产物进行乳酸发酵程序，其中，取一预定量的芽孢乳酸菌，接种量为5％w/v(每毫升有效菌量需达108CFU)，进行通气培养，其中，通气量为0.5VVM，搅拌速率为300rpm、培养温度约为35℃，当发酵时间达72小时后，以高效能液相层析仪监测发酵产物的乳酸含量，在乳酸含量大于600ppm，表示乳酸发酵程序完成，获得一第三发酵产物，其中，该第三发酵产物的该糖份小于5％。
以HPLC检测各该第三发酵产物，结果如下表三所示。
表三：以HPLC检测各该第二发酵产物的酸碱值、总单双糖量、酒精含量、醋酸含量、乳酸含量的结果
| 原料 | pH | 总单双糖 | 酒精含量 | 醋酸含量 | 乳酸含量 |
| 柑橘 | 3.19±0.02 | 0.13％±0.03％ | 0.14％±0.06％ | 2.06％±0.08％ | 739ppm |
| 木瓜 | 3.20±0.10 | 0.19％±0.11％ | 0.27％±0.06％ | 2.12％±0.06％ | 818ppm |
| 凤梨 | 3.19±0.07 | 0.20％±0.09％ | 0.18％±0.03％ | 1.95％±0.06％ | 784ppm |
| 香蕉 | 3.21±0.14 | 0.18％±0.09％ | 0.13％±0.06％ | 1.99％±0.02％ | 889ppm |
| 青梅 | 3.09±0.05 | 0.17％±0.09％ | 0.06％±0.03％ | 2.14％±0.07％ | 816ppm |
| 诺丽果 | 3.33±0.06 | 0.18％±0.06％ | 0.05％±0.03％ | 1.93％±0.05％ | 650ppm |
| 香檬皮 | 3.25±0.13 | 0.21％±0.09％ | 0.08％±0.03％ | 2.00％±0.03％ | 661ppm |
| 葡萄柚 | 3.30±0.17 | 0.16％±0.06％ | 0.09％±0.03％ | 2.07％±0.08％ | 735ppm |
| 奇异果 | 3.29±0.08 | 0.12％±0.08％ | 0.20％±0.04％ | 2.18％±0.04％ | 833ppm |
| 莱姆 | 3.17±0.14 | 0.13％±0.08％ | 0.15％±0.02％ | 2.11％±0.06％ | 726ppm |
| 葡萄 | 3.31±0.15 | 0.27％±0.09％ | 0.15％±0.04％ | 2.00％±0.02％ | 713ppm |
步骤105：分离过滤
通过分离过滤器移除该第三发酵产物中的残渣，以获得一酵素液，其糖份低于5％。
通过上述步骤的组成，本发明所揭低糖蔬果酵素液的制作方法能够有效地产制出低糖份的酵素液，使酵素液的糖度低于原本发酵原料糖度的1/3，并且，所获得的该酵素液后续能够与其他可食用的成份或/及益生菌混合而形成酵素相关产品。
取不同蔬果原料以本发明所揭低糖蔬果酵素液的制作方法进行发酵，在各阶段检测其所得发酵产物的酸碱值，如下表四所示，并将以不同蔬果原料所获得的酵素液以HPLC进行分析糖度、酒精度及酸度，结果如下表五至表九所示，其中，发酵原料为2种以上蔬果时，各蔬果的重量为相同。
表四：不同原料在各阶段所得发酵产物的酸碱值
表五：不同原料制得的酵素液经HPLC分析的结果
表六：不同原料制得的酵素液经HPLC的结果所得其糖度
表七：不同原料制得的酵素液经HPLC分析的结果
表八：不同原料制得的酵素液通过HPLC分析结果得到其酸度
表九：不同原料制得的酵素液经HPLC分析其酒精度的结果
由上表五至九可知，通过本发明所揭低糖蔬果酵素液的制作方法确实能将不同蔬果原料以多种菌种进行多次发酵程序，而能得到不同风味的低糖度发酵液。
又，比较以本发明所揭酵素液与市售酵素液的糖度，结果如下表十所示，其中，表十中的本发明指以本发明所揭方法获得的酵素液，该酵素液的发酵原料为柑橘、木瓜、凤梨、香蕉、奇异果、来姆、葡萄或上述至少一水果与其他蔬果的组合，并且，上述水果的重量大于等于该发酵原料重量的50％。
表十：糖度比较
由表十的结果可清楚得知，本发明低糖蔬果酵素液的制作方法能够有效地大幅降低发酵液的糖度。
Claims (9)
1.一种低糖蔬果酵素液的制作方法，其特征在于，包含下列步骤：
a.取一培养基，具有至少一蔬果原料，其中，该培养基的糖度为15～25度；
b.取一活化的酵母菌，菌种为Saccharomyces cerevisiae，并以0.01～1％w/v的接种量将该酵母菌接种于该培养基上，进行无氧发酵，当发酵至发酵产物的残糖含量低于2％及酒精含量为4.5～5.5％时，得到一第一发酵产物；
c.取一醋酸菌，菌种为Acetobacter aceti，并以1～10％w/v的接种量将该醋酸菌接种于该第一发酵产物上，进行有氧培养，发酵温度为35～40℃，当发酵至发酵产物的醋酸浓度为1.8％～2.5％时，得到一第二发酵产物；
d.取一芽孢乳酸菌，并以5～20％w/v接种量将该乳酸菌接种于该第二发酵产物上，进行通气培养，当发酵至发酵产物的乳酸含量大于600ppm时，得到一第三发酵产物，并该第三发酵产物的糖份低于5％；
e.过滤该第三发酵产物而得到一酵素液。
2.如权利要求1所述低糖蔬果酵素液的制作方法，其特征在于，该发酵原料中的至少一蔬果选自由柑橘、木瓜、凤梨、香蕉、奇异果、来姆、葡萄所组成的群。
3.如权利要求1所述低糖蔬果酵素液的制作方法，其特征在于，当该蔬果的重量大于等于该发酵原料重量的50％时，该酵素液的糖份低于2.5％。
4.如权利要求1所述低糖蔬果酵素液的制作方法，其特征在于，该第二发酵产物的糖度低于第一发酵产物的糖度至少约10度。
5.如权利要求1所述低糖蔬果酵素液的制作方法，其特征在于，该第一发酵产物的酒精度较未发酵前上升至少3％。
6.如权利要求1所述低糖蔬果酵素液的制作方法，其特征在于，该第二发酵产物的醋酸浓度大于2％。
7.如权利要求1所述低糖蔬果酵素液的制作方法，其特征在于，该第二发酵产物的酒精度小于1％。
8.如权利要求1所述低糖蔬果酵素液的制作方法，其特征在于，该第三发酵产物的酒精度小于1％。
9.如权利要求1所述低糖蔬果酵素液的制作方法，其特征在于，该步骤d中的该第三发酵产物的醋酸浓度大于2％。
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