CN104560918A - 一种高效制备低聚甘露糖的复合酶制剂及其应用 - Google Patents

一种高效制备低聚甘露糖的复合酶制剂及其应用 Download PDF

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CN104560918A
CN104560918A CN201410817248.7A CN201410817248A CN104560918A CN 104560918 A CN104560918 A CN 104560918A CN 201410817248 A CN201410817248 A CN 201410817248A CN 104560918 A CN104560918 A CN 104560918A
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唐存多
邬敏辰
阚云超
姚伦广
唐青海
史红玲
黄志刚
焦铸锦
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Nanyang Normal University
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Abstract

本发明公开一种高效制备低聚甘露糖的复合酶制剂，包括含量为100-1000？IU/mL的β-甘露聚糖酶和50-2000？IU/mL的纤维素酶，以复合酶制剂总重量为基准还包括蛋白胨、淀粉、山梨酸钾和木糖醇。所述的纤维素酶为β-内切葡聚糖酶。利用复合酶制剂制备低聚甘露糖的方法，将魔芋精粉、水和复合酶制剂混合，使魔芋精粉的浓度为5-40？g/L，复合酶制剂的加入量使每克魔芋精粉对应有10-80？IU？β-甘露聚糖酶；混合后在？20-70℃的酶解温度下酶解1-10？h。该复合酶制剂比单一酶催化效率高，制剂中还含有酶以外的成分，这些成分与两种酶结合，使酶活残留率高，催化效率更高、更加稳定。

Description

一种高效制备低聚甘露糖的复合酶制剂及其应用
技术领域
本发明属于生物工程技术领域，具体涉及一种高效制备低聚甘露糖的复合酶制剂及其应用。
背景技术
低聚甘露糖是由2-10个甘露糖通过β-1,4糖苷键聚合而成的低聚糖，又称甘露寡糖。它可有效促进生物体内以双歧杆菌为代表的肠道益生菌群的增殖，并具有抑制体内病原菌生长、减少有毒代谢产物产生、防止便秘、保护肝脏、抗肿瘤及增强机体免疫力等多种生理功能，是新一代功能性食品。我国已制定了功能性低聚糖行业标准，并将低聚甘露糖列为其一，但目前国内相应产品的市场占有率仍较低。
魔芋属于天南星科魔芋属的多年生草本植物，是我国南方地区的特有经济作物，其主要成分为魔芋葡甘露聚糖。魔芋葡甘露聚糖主要由甘露糖和葡萄糖组成，它们的摩尔比为1.5:1。目前，在国内魔芋粉主要被当作一种粗纤维食用或用作简单的食品添加剂，附加值不高。而将魔芋粉水解制成低聚甘露糖，可大大拓宽魔芋粉的应用范围，提高原料的附加值，具有重大的经济和社会效益。目前低聚甘露糖的制备通常都是利用高温水解、酸水解或者碱水解，酶法水解虽然也有报道，但条件还不够成熟。酶法生产低聚甘露糖主要受制于生产所用的关键酶的催化效率较低、酶的生产和使用成本较高，以及底物的粘度过大难以实现高浓度的生产等。因此，加大对用于酶法生产低聚甘露糖的复合酶制剂的研究也具有十分重要的意义。
发明内容
本发明的目的是提供一种高效制备低聚甘露糖的复合酶制剂及其应用，该复合酶制剂比单一酶催化效率高，制剂中还含有酶以外的成分，这些成分与两种酶结合，使酶活残留率高，催化效率更高、更加稳定。
本发明实现上述目的采用的技术方案是：一种高效制备低聚甘露糖的复合酶制剂，包括含量为100-1000 IU/mL的β-甘露聚糖酶和50-2000 IU/mL的纤维素酶，以复合酶制剂总重量为基准还包括蛋白胨0.1-4%、淀粉0.1-5%、山梨酸钾0.1-1%和木糖醇0.1-20%。
所述的纤维素酶为β-内切葡聚糖酶。
利用复合酶制剂制备低聚甘露糖的方法，将魔芋精粉、水和复合酶制剂混合，使魔芋精粉的浓度为5-40 g/L，复合酶制剂的加入量使每克魔芋精粉对应有10-80 IU β-甘露聚糖酶；混合后在 20-70℃的酶解温度下酶解1-10 h。
利用复合酶制剂制备低聚甘露糖的方法，将魔芋精粉、水和复合酶制剂混合，使魔芋精粉的浓度为30 g/L，复合酶制剂的加入量使每克魔芋精粉对应有60 IU β-甘露聚糖酶；混合后在 60℃的酶解温度下酶解6 h。
本发明的有益效果
本发明提供的复合酶制剂以耐高温的重组β-甘露聚糖酶为主，辅以不同活性单位的高催化活性的重组β-内切葡聚糖酶对魔芋精粉进行水解比单一酶催化效率高，制剂中还含有酶以外的成分，这些成分与两种酶结合，使酶的催化效率更高，更加稳定，酶活残留率高，且酶的生产和使用成本较低。
附图说明
图1为魔芋精粉水解产物的薄层层析分析。
具体实施方式
一种高效制备低聚甘露糖的复合酶制剂，包括含量为100-1000 IU/mL的β-甘露聚糖酶和50-2000 IU/mL的纤维素酶，以复合酶制剂总重量为基准还包括蛋白胨0.1-4%、淀粉0.1-5%、山梨酸钾0.1-1%和木糖醇0.1-20%。
所述的纤维素酶为β-内切葡聚糖酶。
利用复合酶制剂制备低聚甘露糖的方法，将魔芋精粉、水和复合酶制剂混合，使魔芋精粉的浓度为5-40 g/L，复合酶制剂的加入量使每克魔芋精粉对应有10-80 IU β-甘露聚糖酶；混合后在 20-70℃的酶解温度下酶解1-10 h。
利用复合酶制剂制备低聚甘露糖的方法，将魔芋精粉、水和复合酶制剂混合，使魔芋精粉的浓度为30 g/L，复合酶制剂的加入量使每克魔芋精粉对应有60 IU β-甘露聚糖酶；混合后在 60℃的酶解温度下酶解6 h。
以下结合具体实施例对本发明做进一步说明：
实施例1：
复合酶制剂中各原料的含量为β-甘露聚糖酶100IU/mL和β-内切葡聚糖酶50 IU/mL，以复合酶制剂总重量为基准还添加有蛋白胨4%、淀粉1%、山梨酸钾0.1%和木糖醇10%；复合酶制剂的用量以β-甘露聚糖酶为标准，每克魔芋精粉需要添加β-甘露聚糖酶15 IU；制备低聚甘露糖的原料为魔芋精粉，魔芋精粉的配制溶剂为水，魔芋精粉的浓度为20 g/L，酶解时间为2 h，酶解温度为25℃。在此条件下所得的魔芋精粉的水解率为45.2%，较单一的β-甘露聚糖酶的效果有明显的提高。
实施例2：
复合酶制剂中各原料的含量为β-甘露聚糖酶500IU/mL和β-内切葡聚糖酶1000 IU/mL，以复合酶制剂总重量为基准还添加有蛋白胨2%、淀粉3%、山梨酸钾0.5%和木糖醇15%；复合酶制剂的用量以β-甘露聚糖酶为标准，每克魔芋精粉需要添加β-甘露聚糖酶80 IU；制备低聚甘露糖的原料为魔芋精粉，魔芋精粉的配制溶剂为水，魔芋精粉的浓度为30 g/L，酶解时间为10 h，酶解温度为70℃。在此条件下所得的魔芋精粉的水解率为77.5%，较单一的β-甘露聚糖酶的效果提高得更加显著。
实施例3：
利用所述的复合酶制剂制备低聚甘露糖的工艺条件为：复合酶制剂中各原料的含量为β-甘露聚糖酶1000 IU/mL和β-内切葡聚糖酶1500 IU/mL，以复合酶制剂总重量为基准还添加有蛋白胨2%、淀粉5%、山梨酸钾0.9%和木糖醇20%；复合酶制剂的用量以β-甘露聚糖酶为标准，每克魔芋精粉需要添加β-甘露聚糖酶60 IU；制备低聚甘露糖的原料为魔芋精粉，魔芋精粉的配制溶剂为水，魔芋精粉的浓度为30g/L，酶解时间为6 h，酶解温度为60℃。在此条件下所得的魔芋精粉的水解率为68.4%，较单一的β-甘露聚糖酶的效果提高得更加显著。
试验研究
一、复合酶的稳定性实验
将加有稳定剂的复合酶与没有添加稳定剂的复合酶同时于常温保藏6个月，期间每1个月测定1次复合酶中各组分的残留酶活，计算酶活残留率，其中酶活残留率=残留酶活/初始酶活×100%，结果如表1所示。
表1 添加稳定剂与不添稳定剂复合酶的酶活残留率
二、魔芋精粉水解产物的定性分析
以硅胶板为层析载体，采用2:1:1的丁醇、冰醋酸和水的混合液作为展层剂，0.4 mL苯胺、0.4 g二苯胺和2 mL 85%浓磷酸溶于20 mL丙酮的混合液为显色剂，对实施例3得到的魔芋精粉水解产物进行薄层层析定性分析。由于样品中低聚糖的浓度较低，因此每个样品需点5次，在展层剂中展层5 h后，将硅胶板放置在通风橱中，通风30 min，待展层剂完全挥发掉后用喷壶喷洒显色剂，于室温晾20 min，再于105℃烘3-5 min，最后拍照观察。
结果如图1所示，复合酶的水解效率明显高于单一β-甘露聚糖酶的水解效率，图1中泳道1为未水解的底物，泳道2为单一β-甘露聚糖酶的水解产物，泳道3为复合酶的水解产物。

Claims (4)

1.一种高效制备低聚甘露糖的复合酶制剂，其特征在于：包括含量为100-1000 IU/mL的β-甘露聚糖酶和50-2000 IU/mL的纤维素酶，以复合酶制剂总重量为基准还包括蛋白胨0.1-4%、淀粉0.1-5%、山梨酸钾0.1-1%和木糖醇0.1-20%。
2.如权利要求1所述的一种高效制备低聚甘露糖的复合酶制剂，其特征在于：所述的纤维素酶为β-内切葡聚糖酶。
3.利用如权利要求1所述的复合酶制剂制备低聚甘露糖的方法，其特征在于：将魔芋精粉、水和复合酶制剂混合，使魔芋精粉的浓度为5-40 g/L，复合酶制剂的加入量使每克魔芋精粉对应有10-80 IU β-甘露聚糖酶；混合后在 20-70℃的酶解温度下酶解1-10 h。
4.如权利要求3所述的利用复合酶制剂制备低聚甘露糖的方法，其特征在于：将魔芋精粉、水和复合酶制剂混合，使魔芋精粉的浓度为30 g/L，复合酶制剂的加入量使每克魔芋精粉对应有60 IU β-甘露聚糖酶；混合后在 60℃的酶解温度下酶解6 h。
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CN111518854A (zh) * 2020-04-01 2020-08-11 陕西理工大学 一种魔芋精粉多酶酶解物及其制备方法

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