CN102118964A - 新的甜叶菊变种和制备甜味剂的方法 - Google Patents
新的甜叶菊变种和制备甜味剂的方法 Download PDFInfo
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- CN102118964A CN102118964A CN2009801111403A CN200980111140A CN102118964A CN 102118964 A CN102118964 A CN 102118964A CN 2009801111403 A CN2009801111403 A CN 2009801111403A CN 200980111140 A CN200980111140 A CN 200980111140A CN 102118964 A CN102118964 A CN 102118964A
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Abstract
本发明提供了高纯度蛇菊苷甜味剂。由于通过杂交和选择育种的结果,使用DNA测试来鉴定甜叶菊(Stevia Rebaudiana Bertoni)变种。通过使这些变种杂交,开发出能连续培养特定变种的一种新的变种(保藏号为FERM BP-10870)。通过提取该变种的干燥叶,可以稳定状态获得高浓度的含蛇菊苷的甜味组分。从该提取物开始,可以制备各种有利的蛇菊苷甜味剂。
Description
技术领域
本发明涉及属于甜叶菊(Stevia Rebaudiana Bertoni)变种的植物,所述植物具有极高含量的蛇菊苷,并可由种子连续产生,所有植物都通过使用得自所述植物的种子而产生;和/或涉及从其干燥叶中提取制备甜味剂的方法,并涉及自所述甜味剂制备α-葡糖基蛇菊苷的方法。
背景技术
甜叶菊(Stevia)是一种菊科(Asteraceae)多年生植物,原产于南美巴拉圭,学名:Stevia Rebaudiana Bertoni。栽培甜叶菊用于提取甜味成分,用作天然甜味剂,因为它含有甜度比糖高300倍的成分。
蛇菊苷(C38H60O18)、新蛇菊苷A(C44H70O23)、新蛇菊苷C、D、E、dulcoside A等已知是甜叶菊的甜味成分。广泛栽培的甜叶菊变种含有蛇菊苷(在下文称为ST),是上述甜味成分的主要成分。新蛇菊苷A(在下文称为RA)的含量约为40重量份,ST含量为100重量份,新蛇菊苷C含量略小于RA。然而,主要成分因变种的不同而异。例如,有的变种含有新蛇菊苷C作为其主要成分。
ST广泛用作食品工业的天然甜味剂,因为它的甜度比糖高大于300倍。尽管ST的甜味与糖类似,但与RA相比,已知它具有令人不快的味道,例如作为回味留下的苦味。相比之下,因为RA具有优质甜味,并且比ST的甜度高1.3-1.5倍,所以通常优选含有RA而非ST作为主要成分的甜叶菊甜味剂。
例如,本发明人已经做出了各种改良,即通过对原始变种进行反复杂交和选择,所得甜叶菊变种含有与RA相比极少量的ST,从这些变种的种子,可以容易地培养秧苗(seeding),并可连续培养出甜味成分(参见以下专利文件1和2)。
然而,甜味品质在例如涩味和辛辣味的味道中是特别微妙的,这些是舌头所能感觉到的。与RA相比,甚至ST据信具有令人不快的苦味回味,如果纯化并增加其浓度,去除异味并改善其甜味,因此ST就可用于更广泛的用途。例如,在回味中感觉到ST的甜味,可用于掩蔽咸味食物的咸涩味。另外,ST可用作掩蔽果汁的涩味、有机酸和盐的掩蔽剂,因此,需要ST甜味剂作为不同于RA的甜味剂,用于食品领域。
有方法用于改善甜叶菊提取物的甜味,即通过α-葡糖基化和随后的糖链调整(参见以下专利文件3)。通常,由上述方法制备的含有α-葡糖基蛇菊苷作为主要成分的甜味剂,如果与常见的甜叶菊提取物相比,除ST之外的甜味成分对甜味品质具有较少的干扰。
在资源的有效利用方面,也优选促进ST的使用,因为它是甜叶菊甜味剂的主要成分。
[专利文件1]JP-A-2002-262822
[专利文件2]WO2006/093229
[专利文件3]JP-A-H02-163056
发明内容
本发明要解决的问题
过去,获得高纯度ST的唯一方法就是通过反复重结晶。甜叶菊中除ST之外的任何成分都会导致ST重结晶得率低。因此,有必要开发具有较低RA含量(其是甜叶菊中的第二主要成分)的新的甜叶菊变种,以便降低高纯度ST的制备成本并保持其稳定的得率。
另外,已经考虑到在种子培育中不可能维持甜味成分的恒定,因为在甜叶菊植物育种中,不可能仅通过植株高度或其叶片形状来鉴别变种,而且甜叶菊植物因为自交不亲和而容易杂交。
解决问题的手段
本发明人已经做出了各种改良,即通过对原始变种进行反复杂交和选择,所得甜叶菊变种含有RA的量比ST明显更少。他们从上述甜叶菊变种的种子中成功地开发出新的甜叶菊变种,其中可以容易地培养秧苗(seeding),并可连续培养出甜味成分。
发明效果
自本发明人的新甜叶菊变种干燥叶中得到的提取物具有低RA含量和高ST含量,因此,一次重结晶可获得实际上不含RA的ST甜味剂。或者,可通过酶处理提取物,得到具有改善的甜味品质的ST甜味剂。
实施本发明的最佳方式
本发明的第一实施方案是甜叶菊(Stevia Rebaudiana Bertoni)变种,所述变种含有相对于100重量份的ST不超过8重量份的RA。为了这个目的,通过如下所述的反复杂交和选择,产生含有相对于100重量份ST不超过8重量份RA的属于甜叶菊变种的一种新植物,并通过DNA分析,使用特定引物,对所述变种进行鉴定。
本发明的第二实施方案是制备高纯度ST甜味剂的方法,所述甜味剂含有相对于100重量份的ST不超过8重量份的RA,其中用水或含水溶剂提取上述植物或其干燥叶,然后对所得粗品进行重结晶。
本发明的第三实施方案是制备α-葡糖基蛇菊苷甜味剂的方法,其中用水或含水溶剂提取上述植物或其干燥叶,然后将所得粗品或由粗品重结晶而得到的高纯度ST,与例如环糊精葡糖基转移酶反应,以便通过α-加成而加上葡萄糖。
本发明的第四实施方案是制备α-葡糖基化蛇菊苷甜味剂的方法,其中进行糖链调整,其中由上述第三实施方案所获得的甜味剂通过α-1,4-葡萄糖苷酶例如葡糖淀粉酶处理,以调整所加的糖链。
本发明的第五实施方案是通过RAPD(随机扩增多态性DNA)方法进行DNA分析,使用如下所述的特定引物。
为了达到这些目的,本发明人通过反复杂交和选择进行了各种改良。在这种情况下,用于鉴别所选变种的方法是重要的。如上所述,甜叶菊具有强烈的自交不亲和性,所以在所鉴定的变种之间进行杂交是重要的。为了这一目的,本发明人研究了通过DNA分析来鉴定变种的方法。
RAPD方法是一种DNA分析技术,并且是以下的方法:该方法包括进行PCR(聚合酶链式反应),使用多种引物,扩增在与所用引物得那些相同或类似的序列之间遇到的DNA区,并通过电泳分析所扩增DNA的带型。另外,十六烷基三甲基溴化铵(CTAB)是一种具有长链烷基的季铵碱,可用于分离核酸,因为它与聚阴离子例如核酸形成不溶性复合物。
根据DNA差异来鉴别变种的方法包括:通过CTAB,从植物中分离基因组DNA,除去核糖核酸(RNA),通过PCR方法,使用引物混合物,获得PCR扩增产物,进行电泳并根据所获指纹图谱间的差异来区别PCR扩增产物。对于本发明的植物,如下所述,证实具有位于210bp上游的特征性碱基序列。
在实践中,选择性沉淀的基因组DNA用作原料植物的模板,以下序列用作引物:
正向引物:CACGCGAACTCCTCGACTCGACC
反向引物:GCTGCATGCTTGCATGCATGAAATC
采用以下PCR循环:35次循环,96℃10秒钟,65℃30分钟,72℃30秒钟,再在72℃2分钟。反应之后,所得PCR产物通过6%改性聚丙烯酰胺电泳分离,用SYBR Gold染色并通过蓝光瞬变显示器(transilluminator)观察。结果,在约190bp和210bp检测到显示出多态性的条带,并在所测10个变种中证实F1型的基因型。证实这些条带以良好重现性得以扩增,因此,可以通过特定DNA条带来鉴别植物。
用于制备甜味剂的方法包括:用水和/或含水有机溶剂提取所述植物或其干燥叶,然后浓缩所得提取物;或包括:如上所述进行提取,任选通过使用以下方法除去离子杂质而纯化提取物:例如阳离子交换树脂、阴离子交换树脂或活性炭,用吸附树脂吸附甜味成分,用亲水性溶剂来洗脱,任选通过使用例如阳离子交换树脂、阴离子交换树脂或活性炭进行处理,来浓缩洗脱液,并将其干燥。甜味剂制备方法可包括常规纯化方法,例如脱色,制备高纯度ST的方法可包括常规方法,例如膜分离、醇提取或结晶。另外,结晶方法可使用有机溶剂,例如乙醇或甲醇,其中可任选加入水。其它天然或人工甜味剂、稀释剂等可添加到所得的甜味剂中。
所得甜味剂可用于减低卡路里、减少糖、降低熔点、改善甜味品质并掩蔽糖果、果冻、饮料、饮料粉、即食面、果酱、冷冻甜点、口香糖、日本糖果、保健食品、巧克力、餐桌甜味剂、烘焙糕点、精制食品(delicacies),煮过的海鲜酱产品、乳酸饮料、乳酸菌饮料、咖啡饮料、可可饮料、茶饮料、甜酒、葡萄酒、果子露、谷物、含植物纤维的食物、沙司、酱油、豆酱、醋、调味品、蛋黄酱、番茄酱(ketchup)、咖喱、汤、米制糕点、米制方糕(cubic rice cracker)、面包、饼干、薄脆饼干、薄饼混合料(pancake mix)、罐装水果、罐装蔬菜、肉类产品、煮熟的鱼酱制品、含盐食物、泡菜、调料混合物(seasoning mix)、奢侈食物、化妆品等。还可进一步添加天然高强度甜味剂,例如甘草的甘草甜素、罗汉果(罗汉果)和奇甜蛋白;人工甜味剂,例如环己磺酸钠、糖精钠、阿斯巴甜、三氯半乳蔗糖和纽甜(neotame);和稀释剂,例如低聚糖、糖醇、糖和糊精。
含有高浓度ST的变种的育种方法
通过原始甜叶菊变种的杂交和选择,进行育种。首先,将含有比原始变种更高浓度ST的SS变种杂交,再从所得种子中选择含有相对高浓度ST的SS-1变种。所选变种进一步进行人工杂交,再从所得种子中选择含有较低浓度RA的变种。然后,通过在所选变种内进一步杂交,开发出含有相对于100重量份ST小于5重量份RA的10个变种,然后因缺乏耐寒性而排除掉一个变种之后,称为F1ST-1至F1ST-9的9个变种经鉴别是高ST变种。
将得自这些高ST变种的种子播种,培养植株,分析采自所述植株的干燥叶,证实其甜味成分与高ST变种没有差异。然后,对所述变种的基因进行测序,共享相同基因的变种经鉴别为F1ST变种。
另外,申请人完成了得自本发明F1ST变种的种子的国际保藏(国家先进工业科学技术研究所的国际专利生物保藏中心( InternationalPatent Organism Depository of National Institute of Advanced IndustrialScience and Tech
必要时,本发明的变种可与另一甜叶菊变种杂交。然后,通过以下实施例1所述的选择,可非常容易的得到含有高浓度ST的植物。所有这些植物都包括在可由ST变种的保藏种子得到的植物的范围内。
实施例
以下更详细地说明了育种方法及其特征。本发明并不限于以下育种方法或培养系统。
实施例1
在2001年,从种子中选择含有相对高浓度ST的变种,所述种子通过含有相对高浓度ST的变种杂交而得到,并在Morita KagakuKogyo Co.,Ltd.的Niimi工厂的塑料温室中进行人工杂交。在2002年3月,将所得种子播种在Niimi工厂的塑料温室中,再将萌发的秧苗移栽到盆内,培养秧苗。在5月早期,在施用氮肥、磷肥和钾肥成分各20kg/10之后2周,将高度超过约8cm的500棵秧苗移栽到工厂的育种田中。在7月早期,施用氮肥、磷肥和钾肥成分各10kg/10作为追肥。
在9月早期,通过采集植物并分析其甜味成分,选择具有相对于100重量份ST小于30重量份RA的SS变种株。在2002年,SS变种在Niimi工厂的塑料温室中进行人工杂交。在2003年3月,将所得种子播种在Niimi工厂的塑料温室中,再将萌发的秧苗移栽到盆内,培养秧苗。在5月早期,在施用氮肥、磷肥和钾肥成分各20kg/10之后2周,将高度为约7cm的300棵秧苗移栽到工厂的育种田中。在7月早期,施用氮肥、磷肥和钾肥成分各10kg/10作为追肥。
在9月早期,通过采集植物并分析其甜味成分,选择具有相对于100重量份ST小于15重量份RA的SS-1变种株。在2003年,将所述植株杂交。在2004年3月,将所得种子播种在Niimi工厂的塑料温室中,再将萌发的秧苗移栽到盆内,培养秧苗。在5月早期,在施用氮肥、磷肥和钾肥成分各20kg/10之后2周,将高度约7cm的300棵秧苗移栽到工厂的育种田中。在7月早期,施用氮肥、磷肥和钾肥成分各10kg/10作为追肥。
在9月早期,通过采集植物并分析其甜味成分,具有相对于100重量份ST小于8重量份RA的植株经鉴别为F1ST变种。在2004年,将F1ST变种杂交。在2005年3月,将所得种子播种在Niimi工厂的塑料温室中,再将萌发的秧苗移栽到盆内,培养秧苗。在5月早期,在施用氮肥、磷肥和钾肥成分各20kg/10之后2周,将高度约7cm的1,000棵秧苗移栽到工厂的育种田中。在7月早期,施用氮肥、磷肥和钾肥成分各10kg/10作为追肥。
在9月早期,在收获所有植物并分析其甜味成分之后,证实干燥叶中含有相对于100重量份ST小于8重量份RA。
比较实验1(5月)
为了比较ST变种和另一变种SN,在2006年3月,将得自F1ST变种和SN变种的各1,500粒种子播种在Niimi工厂的塑料温室中,再将萌发的秧苗移栽到盆内,培养秧苗。在5月早期,将每种变种的400棵秧苗生长至与前述相同的高度,并在施用氮肥、磷肥和钾肥成分各20kg/10之后2周,移栽到工厂的育种田中。
在5月中旬,从ST变种和SN变种的各200个茎秆上分离叶子并干燥,用于样品分析。在下列条件下,通过高效液相色谱测定甜味成分。
<高效液相色谱>
柱:LiChrosolv NH2,5μ,4mm×250mm
流速:1.5ml/min
展开溶剂:乙腈∶水=82∶8
波长:210nm
得到以下结果:
[表1]
比较实验2(9月)
在7月中旬,追加一次肥料。在9月早期,在刚刚地面以上收割每一变种的150个茎秆并分离叶子。将叶子干燥,测定甜味成分,用作分析样品。
得到以下结果:
[表2]
在F1ST变种和SN变种中,随着植物的生长,观察到甜味成分含量增加的趋势。然而,在F1ST变种中没有观察到RA含量的大的增加。
基因碱基序列的鉴定
<DNA提取>
通过CTAB方法进行DNA提取。将从每种样品收集到的约0.2g叶子在研钵中用液氮冷冻并用研棒研磨。将研磨过的样品在1.5ml微量管中与0.5ml 2%CTBA溶液(100mM Tris-HCl pH 8.0,20mM EDTApH 8.0,2%CTAB,1.4M NaCl,1%PVP)混合,并在65℃孵育30分钟。加入0.5ml氯仿/异戊醇(24∶1)并搅拌10分钟,然后在1,500rpm离心15分钟,然后将水层移至另一1.5ml微量管中。加入等量的100%异丙醇,并在1,500rpm离心15分钟,然后用75%乙醇漂洗,然后干燥并溶于400μl TE缓冲液中。加入1μl RNase溶液,并在37℃孵育1小时,然后加入等量的TE-饱和苯酚,然后在1,500rpm离心15分钟。将水层移至另一微量管中,并加入1/10量的3M乙酸钠和等量的异丙醇,并混合,然后在1,500rpm离心30分钟。所得沉淀用75%乙醇漂洗2次,干燥并溶于50μl TE缓冲液中,用作DNA样品。
<PCR分析>
用所得DNA样品作为模板,进行PCR分析。PCR循环进行如下:35次循环,96℃10秒钟,65℃30分钟,72℃30秒钟,再在72℃2分钟。反应之后,PCR产物通过6%改性聚丙烯酰胺电泳分离,用SYBRGold染色,并用蓝光瞬变显示器观察。结果,在约190bp和210bp检测到显示出多态性的条带,并在所测10个变种中证实F1ST型的基因型。证实这些条带以良好重现性得以扩增,因此,有效用作鉴别标记。
实施例2:高纯度ST甜味剂的制备
(1)提取
将得自F1ST-1至F1ST-9的每个变种的5月和9月的2g干燥叶混合,制备18g混合物并提取几次,直到20倍稀释水溶液无甜度。将提取物上样到柱子中,该柱子装有20ml阴离子交换树脂[DuoliteA-4]和5ml活性炭颗粒,然后将洗脱液上样到柱子中,该柱子装有100ml吸附树脂[Diaion HP-20],以便吸附甜味成分。用水充分洗涤之后,用300ml甲醇洗脱柱子。将洗脱液真空浓缩并干燥,得到浅黄白色粉末。
(2)重结晶
将上述纯化提取物(5月和9月的F1ST变种)各10g加热并溶于10倍95%甲醇,然后在4℃冷却6天。将所得晶体分离,用冷甲醇洗涤并真空干燥,得到各7.5g和7.4g白色晶体。
为了进行比较,对SN变种进行与上述相同的处理,通过高效液相色谱,采用与实施例1中那些相同的条件,对所得4.6g白色晶体(SN-CRY)进行分析。
纯化提取物的分析结果见表3所示。在该表中,ST代表蛇菊苷,RA代表新蛇菊苷A。
[表3]
变种名称 | ST(%) | RA(%) | RA比率* | %得率(g) |
F1ST 5月 | 93.8 | 0 | 0 | 75%(7.5g) |
F1ST 9月 | 98.4 | 0.1 | 0 | 74%(7.4g) |
SN-CRY | 90.1 | 5.6 | 6 | 46%(4.6g) |
*相对于100重量份ST的RA重量份
通过来自本发明变种F1ST-1至F1ST-9的ST的仅仅一次重结晶,就可得到基本上不含RA的高纯度ST甜味剂。
感官测试1
将得自实施例2的F1ST 5月粉末和SN-CRY粉末制成0.02%溶液,由专门研究甜叶菊甜味剂品质的10名参与者比较其苦味、涩味和甜味品质。
[表4]
尽管对样品的苦味和涩味没有大的差异,但是F1ST变种明显改善了味觉品质。
实施例3:α-葡糖基蛇菊苷甜味剂的制备
将按照实施例2得到的F1ST-9月粉末和按照实施例3得到的粉末各4g以及8g DE(淀粉降解率):10的糊精作为α-葡糖基糖,加热以溶于120ml水,然后冷却至70℃。加入氯化钙,根据底物总量来制备1mmol溶液,并将pH调节至6.0。加入100单位的环糊精葡糖基转移酶,并在70℃反应24小时。然后,将各反应溶液在95℃加热30分钟,使酶失活。
过滤各反应溶液以除去悬浮固体之后,将各滤液上样到装有100ml合成吸附树脂Diaion HP-20的柱子上。用水充分洗涤之后,用300ml甲醇洗脱柱子。将洗脱液上样到装有阴离子交换树脂(amberliteIRA-94)的柱子中,脱盐并脱色,然后真空浓缩并干燥,得到2.6g酶处理的α-F1ST和2.5g酶处理的α-F1ST-CRY α-葡糖基蛇菊苷,为白色粉末。
实施例4:对所加糖链的调整
将相对于固体含量1%的市售葡糖淀粉酶[Glucozyme,NAGASE&CO.,LTD]加入到来自实施例3的1.5g酶处理的α-F1ST和酶处理的α-F1ST-CRY中,并在50℃反应5小时。反应之后,通过在95℃加热30分钟使酶失活。过滤各反应溶液以除去悬浮固体之后,将各滤液上样到装有100ml合成吸附树脂Diaion HP-20的柱子上。用水充分洗涤之后,用300ml甲醇洗脱柱子。将洗脱液上样到装有阴离子交换树脂(amberlite IRA-94)的柱子中,脱盐并脱色,然后真空浓缩并干燥,得到0.7g所加糖链调整的α-F1ST和0.6g所加糖链调整的α-F1ST-CRY白色粉末。
作为比较研究,对在实施例2中得到的F1SN变种的浅黄白色粉末进行与上述相同的处理,以得到酶处理的α-SN和所加糖链调整的α-SN-CRY。
感官测试2
与感官测试1相似,在酶处理的α-F1ST和酶处理的α-SN之间比较了甜味品质。
[表5]
通过α-葡糖基化改善了回味和甜味品质,因为得自本发明的新的甜叶菊变种的ST具有高纯度。
感官测试3
与感官测试1相似,在所加糖链调整的α-ST-CRY和所加糖链调整的α-SN-CRY之间比较了甜味品质。
[表6]
通过α-葡糖基化和糖链调整,ST甜味剂没有变味并改善了回味和甜味品质,因为得自本发明的新的甜叶菊变种的ST具有高纯度。
工业实用性
本发明的甜叶菊的新变种具有低RA含量和高ST含量。因此,可从得自所述新变种的栽培的干燥叶中制备各种有利的蛇菊苷甜味剂。
Claims (11)
1.一种甜叶菊(Stevia Rebaudiana Bertoni)变种植物,所述植物含有相对于100重量份的蛇菊苷不超过8重量份的新蛇菊苷A,其中通过使用RAPD方法的DNA分析,可检测到所述植物具有位于210bp的特征性碱基序列。
2.权利要求1的植物,其中所述植物得自甜叶菊变种(保藏号为FERM BP-10870)的种子。
3.一种用于制备甜味剂的方法,所述方法包括用水或含水溶剂提取权利要求1的植物或其干燥叶。
4.一种用于制备甜味剂的方法,所述方法包括用水或含水溶剂提取权利要求2的植物或其干燥叶。
5.一种从甜味剂获得高纯度蛇菊苷的方法,所述甜味剂通过权利要求3的方法获得,其中所述蛇菊苷含有相对于100重量份的蛇菊苷不超过5重量份的新蛇菊苷,并且纯度超过93%。
6.一种从甜味剂获得高纯度蛇菊苷的方法,所述甜味剂通过权利要求4的方法获得,其中所述蛇菊苷含有相对于100重量份的蛇菊苷不超过5重量份的新蛇菊苷,并且纯度超过93%。
7.一种从甜味剂制备α-葡糖基蛇菊苷的方法,所述甜味剂通过权利要求3的方法获得。
8.一种从甜味剂制备α-葡糖基蛇菊苷的方法,所述甜味剂通过权利要求4的方法获得。
9.一种从甜味剂制备所加糖链调整的α-葡糖基蛇菊苷的方法,所述甜味剂通过权利要求7的方法获得。
10.一种从甜味剂制备所加糖链调整的α-葡糖基蛇菊苷的方法,所述甜味剂通过权利要求8的方法获得。
11.一种通过DNA分析来选择权利要求1的甜叶菊变种的方法,所述DNA分析使用RAPD方法,其中以下序列用作引物:
正向引物:CACGCGAACTCCTCGACTCGACC
反向引物:GCTGCATGCTTGCATGCATGAAATC。
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CN105050388A (zh) * | 2013-03-15 | 2015-11-11 | 嘉吉公司 | 具有提高的莱苞迪苷d含量的甜叶菊植物 |
CN110063257A (zh) * | 2015-01-23 | 2019-07-30 | 谱赛科美国公司 | 甜叶菊新品种及高rd、rm含量甜菊糖苷的制备 |
EP4033888A4 (en) * | 2019-09-24 | 2023-12-13 | PureCircle USA Inc. | STEVIA CULTIVAR '320032' WITH SUPER HIGH REBAUDIOSIDE CONTENT |
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US8318459B2 (en) | 2011-02-17 | 2012-11-27 | Purecircle Usa | Glucosyl stevia composition |
CN108464425B (zh) | 2011-01-28 | 2021-10-01 | 泰特&莱尔组分美国公司 | 甜叶菊提取物和甜味组合物 |
US8962698B2 (en) | 2011-01-28 | 2015-02-24 | Tate & Lyle Ingredients Americas Llc | Rebaudioside-mogroside V blends |
US9603373B2 (en) * | 2011-02-17 | 2017-03-28 | Purecircle Sdn Bhd | Glucosyl stevia composition |
GB201217700D0 (en) | 2012-08-01 | 2012-11-14 | Tate & Lyle Ingredients | Sweetener compositions containing rebaudioside B |
USPP27937P3 (en) | 2014-11-30 | 2017-04-25 | S&W Seed Company | Stevia plant named ‘SW 107’ |
USPP27815P3 (en) | 2014-12-08 | 2017-03-28 | S&W Seed Company | Stevia plant named ‘SW 201’ |
USPP28373P3 (en) | 2015-11-17 | 2017-09-12 | S&W Seed Company | Stevia plant named ‘SW 129’ |
USPP28977P3 (en) | 2016-03-31 | 2018-02-20 | S&W Seed Company | Stevia plant named ‘SW 227’ |
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JPS545070A (en) * | 1977-06-13 | 1979-01-16 | Hayashibara Biochem Lab | Production of sweetening agent |
JPS54147976A (en) * | 1978-05-12 | 1979-11-19 | Morita Kagaku Kogyo | Improving of solubility of stevia extracted and refined substance |
JPH02163056A (ja) | 1988-12-16 | 1990-06-22 | Sanyo Kokusaku Pulp Co Ltd | 高甘味糖付加ステビア甘味料及びその製法 |
JPH10271928A (ja) * | 1997-01-30 | 1998-10-13 | Morita Kagaku Kogyo Kk | ステビア・レバウディアナ・ベルトニーに属する新植物 |
JP2002262822A (ja) | 1997-01-30 | 2002-09-17 | Morita Kagaku Kogyo Kk | 種子栽培可能なステビア品種の植物体から得られる甘味料 |
US5972120A (en) * | 1997-07-19 | 1999-10-26 | National Research Council Of Canada | Extraction of sweet compounds from Stevia rebaudiana Bertoni |
US6255557B1 (en) * | 1998-03-31 | 2001-07-03 | Her Majesty The Queen In Right Of Canada As Represented By The Ministerof Agriculture And Agri-Food Canada | Stevia rebaudiana with altered steviol glycoside composition |
JP2002171930A (ja) * | 2000-12-08 | 2002-06-18 | Morita Kagaku Kogyo Kk | 甘味料組成物 |
JP4776107B2 (ja) * | 2001-07-02 | 2011-09-21 | 守田化学工業株式会社 | ステビア品種のdna鑑定による識別 |
MX2007010819A (es) | 2005-03-04 | 2008-01-14 | Morita Kagaku Kogyo | Edulcorante de stevia. |
-
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CN105050388A (zh) * | 2013-03-15 | 2015-11-11 | 嘉吉公司 | 具有提高的莱苞迪苷d含量的甜叶菊植物 |
CN105050388B (zh) * | 2013-03-15 | 2019-08-13 | 嘉吉公司 | 具有提高的莱苞迪苷d含量的甜叶菊植物 |
CN110063257A (zh) * | 2015-01-23 | 2019-07-30 | 谱赛科美国公司 | 甜叶菊新品种及高rd、rm含量甜菊糖苷的制备 |
EP4033888A4 (en) * | 2019-09-24 | 2023-12-13 | PureCircle USA Inc. | STEVIA CULTIVAR '320032' WITH SUPER HIGH REBAUDIOSIDE CONTENT |
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KR20100106509A (ko) | 2010-10-01 |
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US20110023192A1 (en) | 2011-01-27 |
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