CN112839525A - 塔格糖和半乳糖糖浆 - Google Patents

塔格糖和半乳糖糖浆 Download PDF

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CN112839525A
CN112839525A CN201980067240.4A CN201980067240A CN112839525A CN 112839525 A CN112839525 A CN 112839525A CN 201980067240 A CN201980067240 A CN 201980067240A CN 112839525 A CN112839525 A CN 112839525A
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mixture
galactose
tagatose
composition
epimerization
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贾瓦尼·斯波纳蒂
卢瓦纳·瓦尼奥利
雅格布·奇尼
西尔维亚·柏尔吉奥里尼
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Inarco Co Co ltd
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Abstract

本发明描述了作为主要组分的塔格糖和半乳糖连同少量的其它次要产物诸如甘油、低聚糖和其它糖的糖浆。

Description

塔格糖和半乳糖糖浆
发明领域
本发明涉及甜味剂和益生元的领域。
背景
塔格糖是己糖酮类单糖(hexose ketonic monosaccharide)，果糖的异构体。塔格糖是稀有糖，如果乳制品经历加热，可以在乳制品中发现少量的塔格糖。
尽管相对于蔗糖具有等于92％的增甜能力，但是塔格糖提供了减少的热量摄入(38％)，其不是生龋齿的，并且因此其还在糖果工业的焙烤产品的制备中找到作为代替普通的食用糖(table sugar)的甜味剂的应用。
塔格糖具有抗高血糖作用，因为其通过增加负责糖原中葡萄糖的转移的葡萄糖激酶的活性来达成控制餐后血糖水平。其还对肠道中参与碳水化合物的降解的一些酶具有抑制作用，导致它们的吸收减少。
已经进行了关于由塔格糖的摄入引起的血糖指数(glycemic index)的降低的作用的研究(Mark Ensor等人，“Effect of Three Low-Doses of D-Tagatose on Glycemiccontrol Over Six Months in Subjects with Mild Type 2Diabetes Mellitus withDiet and Exercise”J Endocrinol Diabetes Obes.2014年10月；2(4):1057)。
因此，塔格糖在2型糖尿病的治疗中是有用的，已经对其进行了临床研究(ClinicanTrials.gov,NCT00955747,首次发布：2009年8月10日)。
然而，呈结晶形式的塔格糖在食品、运动饮料等中作为甜味剂且作为益生元的使用受到产品的成本的限制，其相对于其它合成分子或提取分子通常不是很有竞争力。
半乳糖是一种简单的糖，是葡萄糖的差向异构体。其在人体中少量产生，而其大部分主要通过摄入含有乳糖二糖的牛奶和乳制品而随饮食引入，所述乳糖二糖通过乳糖酶被分解成葡萄糖和半乳糖。
乳糖是在婴儿的喂养中存在最多的糖，作为生长中的生物体，婴儿需要具有可用的有效的能量来源，并且此外，存在由乳糖产生的半乳糖参与髓磷脂形成的过程的实验证据(Ravera S,Bartolucci M,Cazia D,Morelli A,Panfoli I,“Galactose and Hexose 6-Posphate Dehydrogenase Support the Myelin Metabolic Role”,PARIPEX Indianjournal of research 2015,4(9),第21-24页)。
关于半乳糖对中枢神经系统的积极作用用于治疗退行性疾病诸如例如阿尔茨海默病，已经进行了多种研究(“Therapeutic effect of oral galactose treatment inrat model of sporadic Alzheimer’s”Alzheimer’s&Dementia-The Journal of theAlzheimer’s Association disease,2014年7月，第10卷，第4期，补充页P464)。
存在以下的实验证据：半乳糖和抗氧化剂的摄入还可以用于治疗多发性硬化，尤其是在疾病的发作的早期阶段(Isabella Pandolfi,等人,“Missed evolution ofdemyelinizing brain during supplementation with natural compounds:A casereport”,Medical Research Archives，第4卷，第1期，2016年4月)。
对于半乳糖作为食品补充剂在治疗先天性糖基化紊乱(congenitalglycosylation disorder)中(ClinicanTrials.gov,NCT02955264,首次发布：2016年11月4日)和在治疗2型糖尿病中(ClinicanTrials.gov,NCT01776099，首次发布：2013年1月25日)的用途的临床研究正在进行中。
低聚糖对健康的许多有益性质，所述低聚糖诸如低聚半乳糖(galacto-oligosaccharide)，其是由于乳糖酶(β-半乳糖苷酶)的作用形成的，所述乳糖酶除了对乳糖具有水解活性之外，还具有将半乳糖单元以可变数量加入到乳糖的合成作用。
低聚半乳糖具有促进肠道中微生物(主要是双歧杆菌(bifidobacteria))的生长的益菌作用，并且根据几项研究，它们可能抑制潜在致病微生物的生长(Daniele Garrido等人,“Utilization of galactooligosaccharides by bifidobacterium longumsubsp.Infantis isolates”,Food Microbiol.2013年4月33(2)；262-270)。
本发明的目的是提供基于塔格糖和半乳糖的糖浆及其制备方法。
发明概述
本发明借助于包含以下的组合物解决了前述问题：
Figure BDA0003015857510000031
其中塔格糖/半乳糖比率等于1.0-1.6，其中％是按干组合物的重量计，所述组合物呈以58°-62°白利糖度(brix)的糖量计浓度(saccharometric concentration)的糖浆形式。
因此，本发明的目的是作为主要组分的塔格糖和半乳糖连同少量的其它次要产物诸如甘油、低聚糖和其它糖的糖浆。本发明的组合物使得有可能避免经历塔格糖的结晶(所述经历结晶不可避免地导致结晶母液中的产品的损失)，以及允许缩短生产时间并提高生产率，有益于最终成本。
因此，呈糖浆形式的组合物的优点是明显的，然而应当强调，含糖糖浆的一个问题是，取决于纯度和储存条件，它们倾向于结晶，但是在本发明的组合物的情况下，已经令人惊讶地发现塔格糖和半乳糖之间等于1.0-1.6的比率防止了这种情况的发生，这从商业角度和产品的使用来看具有毫无疑问的优点。
然而，本发明的组合物除了提供塔格糖的摄入之外，还允许其它物质被引入饮食中，诸如半乳糖和低聚半乳糖，由于提及的原因，所述其它物质可以协同地作用，这也为健康提供了积极的贡献。由于塔格糖和半乳糖的有益效果，本发明的糖浆可以被用于制备功能性食品、医疗食品、运动饮料、果汁、酸奶、食品补充剂以及用于糖果工业中。
此外，本发明的目的是用于制备前述糖浆的工艺，所述工艺包括：
i.使乳糖经历通过乳糖酶的酶促水解，以获得包含葡萄糖和半乳糖的混合物；
ii.使包含葡萄糖和半乳糖的混合物与至少一种食用酵母接触，以进行去葡萄糖化(deglucosation)并且获得去葡萄糖化的混合物；
iii.使去葡萄糖化的混合物经历碱性差向异构化，以进行半乳糖向塔格糖的差向异构化并且获得差向异构化的混合物；
iv.使差向异构化的混合物与至少一种离子交换树脂接触，以进行去离子并且获得去离子的混合物；
v.任选地使去离子的混合物经历纳滤，以获得纳滤的混合物；
vi.任选地使去离子的混合物，或任选地所述纳滤的混合物，经历反渗透，以获得渗透渗余物；
vii.使去离子的混合物、或任选地纳滤的混合物、或任选地渗透渗余物，经历陶瓷超滤，以获得超滤的混合物；
viii.使超滤的混合物达到高达58°-62°白利糖度的浓度，以获得糖浆。
附图简述
图1-根据本发明的组合物在磺酸柱(sulphonic column)上的HPLC色谱图的实例。
图2-根据本发明的组合物在胺柱(amine column)上的HPLC色谱图的实例。
发明详述
本发明的糖浆优选地具有等于1.1-1.5、更优选地1.2-1.4的塔格糖/半乳糖比率。
本发明的糖浆优选地具有59°-61°白利糖度的糖量计浓度。
本发明的糖浆优选地具有3.0-3.5的pH。
优选地，本发明的组合物包含：
Figure BDA0003015857510000051
根据本发明的工艺，原料优选地是呈结晶形式的乳糖一水合物。可选择地，还可以使用其它来源的乳糖，诸如例如乳清。
乳糖的酶促水解(i)使用多种来源的商业乳糖酶来进行；例如且优选地：来自乳酸克鲁维酵母(K.Lactis)、脆壁克鲁维酵母(K.Fragilis)、米曲霉(A.oryzae)、黑曲霉(A.niger)、大肠杆菌(E.coli)、嗜热脂肪芽孢杆菌(B.stearothermophilus)、环状芽孢杆菌(B.circulans)的乳糖酶，在本发明中更优选地使用来自米曲霉的酶。
乳糖酶可以以游离形式和在多种类型的固体载体上的固定形式两者来使用，例如且优选地固定在合成树脂、藻酸盐珠、合成膜或棉纤维上，优选地固定酶在本发明中在聚苯乙烯合成树脂上使用，更优选地如在专利申请WO2014006606中所描述的固定酶。
乳糖的酶促水解(i)反应在包括在5℃和60℃之间、优选地在52℃的温度并且在4.0-9.0、优选地5.1-5.5的pH，保持乳糖溶液在柱上再循环下进行持续包括在1小时和48小时之间、优选地20小时的时间。
根据本发明，通过加入食用酵母，优选地冻干的啤酒酵母(酿酒酵母(Scerevisiae))，使包含被水解成葡萄糖和半乳糖的乳糖的溶液经历去葡萄糖化步骤(ii)，直到获得葡萄糖浓度≤0.25％。去葡萄糖化优选地在吹入空气下，通过在包括在25℃和40℃之间、优选地在30℃和37℃之间、更优选地在35℃的温度，在4.0-9.0、优选地6.0-7.0的pH保持在搅拌下来进行持续至少4小时的时间。
在去葡萄糖化的溶液中存在的半乳糖通过经由加入碱性物质的差向异构化(iii)被转化为塔格糖，所述碱性物质例如且优选地氢氧化钠、氢氧化钾或氢氧化钙，更优选地根据本发明使用氢氧化钙。碱性物质优选地以每摩尔半乳糖碱性物质摩尔数的0.1-1.0、优选地0.4-0.8、更优选地0.6的摩尔比率使用。差向异构化(iii)优选地在0℃-30℃、优选地5℃-25℃、更优选地10℃-20℃的温度保持在搅拌下进行持续至少10分钟、优选地4小时的时间。
差向异构化反应(iii)在结束时通过加入优选地选自由盐酸、磷酸、硫酸组成的组的酸来中和，更优选地使用在水中的30％-50％的硫酸。在用酸中和之后，悬浮液优选地被离心以分离沉淀的硫酸钙和来自去葡萄糖化步骤的残留的酵母。
在中和和离心之后获得的溶液通过在一对离子交换树脂上的通过而被去离子(iv)，所述一对离子交换树脂优选地是强阳离子型树脂(诸如例如Rohm and HaasAmberliteTM.200、Rohm and Haas AmberliteTM.IR120、Rohm and Haas AmberliteTM.FPC 23和DowexTM MonosphereTM 88)，接着是弱阴离子型树脂(诸如例如：
Figure BDA0003015857510000061
AmberliteTM FPA55、DowexTM MonosphereTM 66、Rohm and Haas AmberliteTM.IRA 96、
Figure BDA0003015857510000062
A 120S和
Figure BDA0003015857510000063
A109)。
然后，可以任选地但优选地使去离子的混合物经历纳滤(v)，以便至少部分地从上面的二聚体中去除低聚物组分(oligomeric component)。所述纳滤优选地在螺旋卷式膜(spiral wound membrane)上进行，所述螺旋卷式膜选自例如由Dow FilmtecTM NF270-4040、Koch Membrane System TFC-SR2或回收渗透物的类似物组成的组，所述渗透物然后任选地且优选地为了至少部分地去除甘油，在反渗透膜上经历反渗透处理(vi)，所述反渗透膜例如且优选地选自由Es.Dow FilmtecTM BW30-4040组成的组。
根据本发明，在反渗透步骤(vi)中获得的渗余物在优选地300000Da截止值的陶瓷膜上通过超滤(vii)被澄清化，回收渗透物，该渗透物经历浓缩(viii)。在优选地通过加入有机酸，将pH校正在包括在3.0和3.5之间的值之后，将糖浆浓缩直到获得具有60±2°白利糖度的糖量计浓度的糖浆。所述有机酸优选地选自由柠檬酸、乳酸、乙酸组成的组。优选地，用于校正pH的有机酸是柠檬酸，更优选地按重量计40％-60％的柠檬酸的水溶液。
根据以下实施方案的实施例可以更好地理解本发明。
实验部分
HPLC方法：
Perkin Elmer 200系列色谱仪，具有带恒温单元的折射率检测器。
在磺酸柱上分析：带有前置柱的Transgenomic ICE-SEP ION 300柱。温度45℃，流量0.4ml/min，洗脱液硫酸0.015N。
在胺柱上分析：Thermo ScientificTM HypersilTM APS-2。温度40℃，流量1.1ml/min，流动相＝乙腈+磷酸二氢钠二水合物1.45克/升。
实施例1：工业规模的乳糖的酶促水解和去葡萄糖化的溶液的获得。
a)在合成树脂上的固定酶的制备：
在设置有恒温装置(thermostatation)的10m3钢制夹套反应器中，装载750升的Purolite A120S树脂。将树脂用三个750升等分试样的饮用水分别洗涤。加入480升的处于pH 5的100mM乙酸钠溶液和55kg的50％戊二醛溶液。将整体在25℃在搅拌下保持持续30小时，这之后将树脂用三个1000升等分试样的饮用水分别洗涤。加入2000升的处于pH 5的100mM乙酸钠溶液和30kg的来自米曲霉的乳糖酶。将整体在25℃在搅拌下保持持续65小时。在已经经过这段时间之后，将树脂用三个2000升等分试样的饮用水分别洗涤。
b)乳糖的酶促水解：
将2000kg的呈结晶形式的乳糖一水合物溶解在10m3钢制反应器中的8000升的饮用水中，该钢制反应器设置有搅拌和恒温夹套。使反应器的内部温度达到53℃，并且通过加入38％硫酸使pH达到5.39。
将乳糖溶液以2400升/小时的流量通过包含600升树脂(其中乳糖酶如上文实施例1a中所描述的被固定)的柱再循环持续20小时。
在磺酸柱上的分析结果：
半乳糖(HPLC) 9.023％
葡萄糖(HPLC) 9.228％
乳糖(HPLC) 0.492％
葡萄糖/半乳糖 1.02
在已经经过这段时间之后，将包含葡萄糖和半乳糖的溶液转移到20m3的钢制夹套反应器中，该反应器设置有搅拌和空气吹入系统。使温度达到35℃，并且向溶液中加入12kg的冻干的啤酒酵母和100ml的消泡剂(Silifood 1600)。
将整体在充入空气的情况下在35±2℃在搅拌下保持持续10小时。在已经经过这段时间之后，通过加入9升的30％氢氧化钠使pH回到6.8。在pH校正结束时，将10kg的冻干的啤酒酵母引入反应器中，并且在吹入空气的情况下在搅拌下将发酵保持持续另外的10小时。在已经经过这段时间之后，通过加入15升的30％氢氧化钠将pH调节到6.6，并且引入10kg的冻干的啤酒酵母。在已经经过另外的15小时之后，将获得的去葡萄糖化的溶液冷却至约5℃。
在磺酸柱上的分析结果：
半乳糖(HPLC) 8.058％
葡萄糖(HPLC) 0.013％
乳糖(HPLC) 0.491％
葡萄糖/半乳糖 0.2
实施例2：从实施例1的去葡萄糖化的溶液开始，在40℃用相对于半乳糖摩尔数的50％摩尔的氢氧化钙进行实验室规模的差向异构化。
将250g的根据实施例1制备的去葡萄糖化的半乳糖溶液引入到恒温的且设置有搅拌棒的1升玻璃反应器中，并且与4.16g的氢氧化钙(通风的石灰(ventilated lime))合并，将整体在搅拌下保持在40℃的温度。
样品在以下时间之后被采集用于在磺酸柱上的HPLC分析：120min、240min、360min。
结果在下表中表示：
Figure BDA0003015857510000091
实施例3：从实施例1的去葡萄糖化的溶液开始，在40℃用相对于半乳糖摩尔数的60％摩尔的氢氧化钙进行实验室规模的差向异构化。
将250g的根据实施例1制备的去葡萄糖化的半乳糖溶液引入到恒温的且设置有搅拌棒的1升玻璃反应器中，并且与5.0g的氢氧化钙(通风的石灰)合并，将整体在搅拌下保持在40℃的温度。
样品在以下时间之后被采集用于在磺酸柱上的HPLC分析：120min、240min、360min。
结果在下表中表示：
Figure BDA0003015857510000092
实施例4：从实施例1的去葡萄糖化的溶液开始，在30℃用相对于半乳糖摩尔数的50％摩尔的氢氧化钙进行实验室规模的差向异构化。
将250g的根据实施例1制备的去葡萄糖化的半乳糖溶液引入到恒温的且设置有搅拌棒的1升玻璃反应器中，并且与4.16g的氢氧化钙(通风的石灰)合并，将整体在搅拌下保持在30℃的温度。
样品在以下时间之后被采集用于在磺酸柱上的HPLC分析：120min、280min、350min。
结果在下表中表示：
Figure BDA0003015857510000101
实施例5：从实施例1的去葡萄糖化的溶液开始，在30℃用相对于半乳糖摩尔数的60％摩尔的氢氧化钙进行实验室规模的差向异构化。
将250g的根据实施例1制备的去葡萄糖化的半乳糖溶液引入到恒温的且设置有搅拌棒的1升玻璃反应器中，并且与5.0g的氢氧化钙(通风的石灰)合并，将整体在搅拌下保持在30℃的温度。
样品在以下时间之后被采集用于在磺酸柱上的HPLC分析：120min、280min、350min。
结果在下表中表示：
Figure BDA0003015857510000102
实施例6：从实施例1的去葡萄糖化的溶液开始，在25℃用相对于半乳糖摩尔数的50％摩尔的氢氧化钙进行实验室规模的差向异构化。
将250g的根据实施例1制备的去葡萄糖化的半乳糖溶液引入到恒温的且设置有搅拌棒的1升玻璃反应器中，并且与4.16g的氢氧化钙(通风的石灰)合并，将整体在搅拌下保持在25℃的温度。
样品在以下时间之后被采集用于在磺酸柱上的HPLC分析：120min、280min、350min、470min、590min、710min
结果在下表中表示：
Figure BDA0003015857510000111
实施例7：从实施例1的去葡萄糖化的溶液开始，在25℃用相对于半乳糖摩尔数的60％摩尔的氢氧化钙进行实验室规模的差向异构化。
将250g的根据实施例1制备的去葡萄糖化的半乳糖溶液引入到恒温的且设置有搅拌棒的1升玻璃反应器中，并且与5.0g的氢氧化钙(通风的石灰)合并，将整体在搅拌下保持在25℃的温度。
样品在以下时间之后被采集用于在磺酸柱上的HPLC分析：120min、280min、350min、470min、590min、710min。
结果在下表中表示：
Figure BDA0003015857510000112
Figure BDA0003015857510000121
实施例8：从实施例1的去葡萄糖化的溶液开始，以工业规模获得塔格糖/半乳糖糖浆。
碱性差向异构化：
将9100kg的根据实施例1制备的去葡萄糖化的半乳糖溶液引入到设置有搅拌和恒温夹套的10m3钢制反应器中。在30％的饮用水悬浮液中加入192.4kg的氢氧化钙(氢氧化钙的量代表相对于半乳糖60％的摩尔比率)。在加入之后，将温度在15±5℃在搅拌下保持持续4小时。在已经经过这段时间之后，通过加入590升的38％硫酸，将温度保持在低于45℃，使悬浮液的pH达到2.5。然后将温度降低至25℃，并且通过8次在350rpm离心25分钟来分离沉淀的硫酸钙。
在磺酸柱上的分析结果：
Figure BDA0003015857510000122
在离子交换树脂上去离子：
将在先前的离心步骤中获得的溶液在一对离子交换树脂(4000升的强阳离子型树脂AmberliteTM FPC 23和4000升的弱阴离子型树脂AmberliteTM FPA55)上以2000升/小时的流量去离子，从树脂中收集洗脱的产物，直到糖浓度≥0.5°白利糖度且电导率≤50μs/cm。
纳滤：
然后使用由12个膜(
Figure BDA0003015857510000123
FILMTECHTM NF 270 40/40)组成的系统在约30巴的压力使去离子的溶液经历纳滤步骤。
反渗透：
使用由12个膜(
Figure BDA0003015857510000131
FILMTECHTM BW30-400)组成的系统在约10巴的压力使纳滤的渗透物经历反渗透步骤，将渗余物浓缩直到10°白利糖度的糖浓度。
陶瓷超滤：
将浓缩的溶液通过在300000Da截止值的陶瓷膜上、以2000升/小时的渗透流量和9000升/小时的渗余物再循环的切向超滤步骤(tangential ultrafiltration step)来澄清化。将渗余物浓缩直到约300升，并且分别用150升的饮用水洗涤3次。
浓缩：
将来自先前的超滤步骤的澄清化的渗透物转移到设置有搅拌、恒温夹套和冷凝器的5000升钢制反应器中。
通过加入1.6升的在水中的50％柠檬酸使pH达到3.0，并且将溶液在真空下在约50℃的温度浓缩直到60°白利糖度的糖量计浓度。
结果：
将在工艺结束时获得的糖浆在磺酸柱和胺柱两者上进行分析，胺柱用于确定乳糖含量(还参见图1和图2中的HPLC迹线(HPLC trace)),并且结果在下表中表示：
Figure BDA0003015857510000132
Figure BDA0003015857510000141

Claims (11)

1.一种组合物，包含：
Figure FDA0003015857500000011
其中塔格糖/半乳糖比率等于1.0-1.6，其中％是按干组合物的重量计，所述组合物呈以58°-62°白利糖度的糖量计浓度的糖浆形式。
2.根据权利要求1所述的组合物，其中所述塔格糖/半乳糖比率等于1.1-1.5。
3.根据权利要求1-2中任一项所述的组合物，具有59°-61°白利糖度的糖量计浓度。
4.根据权利要求1-3中任一项所述的组合物，具有3.0-3.5的pH。
5.根据权利要求1-4中任一项所述的组合物，包含以下或由以下组成：
Figure FDA0003015857500000012
Figure FDA0003015857500000021
6.一种用于制备根据权利要求1-5中任一项所述的糖浆的工艺，所述工艺包括：
i.使乳糖经历通过乳糖酶的酶促水解，以获得包含葡萄糖和半乳糖的混合物；
ii.使包含葡萄糖和半乳糖的所述混合物与至少一种食用酵母接触，以进行去葡萄糖化并且获得去葡萄糖化的混合物；
iii.使所述去葡萄糖化的混合物经历碱性差向异构化，以进行半乳糖向塔格糖的差向异构化并且获得差向异构化的混合物；
iv.使所述差向异构化的混合物与至少一种离子交换树脂接触，以进行去离子并且获得去离子的混合物；
v.任选地使所述去离子的混合物经历纳滤，以获得纳滤的混合物；
vi.任选地使所述去离子的混合物，或任选地所述纳滤的混合物，经历反渗透，以获得渗透渗余物；
vii.使所述去离子的混合物、或任选地所述纳滤的混合物、或任选地所述渗透渗余物，经历陶瓷超滤，以获得超滤的混合物；
viii.使所述超滤的混合物达到高达58°-62°白利糖度的浓度，以获得根据权利要求1-5中任一项所述的糖浆。
7.根据权利要求6所述的工艺，其中原料是呈结晶形式的乳糖一水合物。
8.根据权利要求6-7中任一项所述的工艺，其中乳糖的酶促水解(i)反应在包括在5℃和60℃之间的温度并且在pH 4.0-9.0、保持乳糖溶液在柱上再循环下进行持续包括在1小时和48小时之间、优选地20小时的时间，其中所述柱包含来自乳酸克鲁维酵母(K.Lactis)、脆壁克鲁维酵母(K.Fragilis)、米曲霉(A.oryzae)、黑曲霉(A.niger)、大肠杆菌(E.coli)、嗜热脂肪芽孢杆菌(B.stearothermophilus)或环状芽孢杆菌(B.circulans)的固定的乳糖酶。
9.根据权利要求6-8中任一项所述的工艺，其中所述去葡萄糖化在吹入空气下、保持在包括在25℃和40℃之间的温度在pH 4.0-9.0在搅拌下进行持续至少4小时的时间。
10.根据权利要求6-9中任一项所述的工艺，其中所述差向异构化(iii)通过加入相对于半乳糖摩尔数的0.1-1.0的氢氧化钙摩尔比率而发生；所述差向异构化(iii)在0℃-30℃的温度在搅拌下进行持续至少10分钟、优选地4小时的时间；其中在差向异构化反应(iii)结束时，所述差向异构化反应(iii)通过加入在水中的30％-50％的硫酸来中和；在用所述酸中和之后，将悬浮液离心以分离沉淀的硫酸钙和来自所述去葡萄糖化步骤的残留的酵母。
11.根据权利要求1-5中任一项所述的组合物在制备功能性食品、医疗食品、运动饮料、果汁、酸奶、食品补充剂中和在糖果工业中的用途。
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