CN106929187B - 一种冷水低温洗涤剂及其制备方法 - Google Patents
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Abstract
本发明提供了一种冷水低温洗涤剂及其制备方法,具体地本发明提供的冷水低温洗涤剂包括低温蛋白酶、酶稳定剂、和表面活性剂,其中所述低温蛋白酶含量为10000u/ml‑300000u/ml,酶稳定剂含量为4%‑10%(w/v),表面活性剂含量为2%‑12%(v/v)。本发明的冷水低温洗涤剂在常温下低温酶的酶活稳定,克服了低温酶在常温下不稳定的缺陷,能够显著延长洗涤剂保质期。
Description
技术领域
本发明属于生物技术领域,具体地说,本发明涉及一种冷水低温洗涤剂及其制备方法。
背景技术
相比于传统洗涤剂,以酶作为助剂的洗涤剂称之为加酶洗涤剂,加酶洗涤剂具有更强的去污能力,因而应用范围更广。而且与含磷洗涤剂相比更为环保,因而,可以取代含磷洗涤剂满足人们的日常和工业需求。
蛋白酶是目前应用最多的酶,目前使用最多的蛋白酶多数为中温蛋白酶,最适酶活在50℃左右。低温酶是指催化温度在30℃左右,在0℃左右仍有一定催化效率的酶,主要由耐低温菌产生。低温蛋白酶最适酶活温度低,在低温下具有更高的催化效率,与中温蛋白酶相比具有无法取代的优越性。自美国华盛顿大学的海洋微生物学家开发的嗜低温蛋白酶应用于洗涤剂工业以来,加酶洗涤剂在冷水洗涤中发挥了越来越显著的效益。
但是低温蛋白酶在常温下的稳定性较差,在加酶洗涤剂中,酶通常不稳定,极性的溶剂、缓冲剂及其各种阴阳离子表面活性剂组分可能改变酶蛋白的高级结构。因此,本领域技术人员致力于开发具有稳定的酶活性的冷水低温洗涤剂。
发明内容
本发明的目的在于提供一种冷水低温洗涤剂及其制备方法。
在研究中本发明人意外发现,使用过的实验用低温蛋白酶在长期放置条件下酶活表现的更为稳定。在此基础上,本发明人进一步研究,发现某些短肽能够作为低温蛋白酶的可逆性抑制剂,对低温蛋白酶起到稳定效果。因此,本发明提供了一种安全、有效、无毒的低温蛋白酶稳定剂,可以避免普通含硼稳定剂的生殖毒性问题,提高了液体低温蛋白酶储存稳定性。
本发明根据发酵浓缩酶配方的特点,设计酶稳定剂,能明显降低生产加工和储存中形成的沉淀和延长液体低温蛋白酶货架期。
本发明的第一方面,提供了一种冷水低温洗涤剂,包括低温蛋白酶、酶稳定剂、和表面活性剂;其中,所述低温蛋白酶含量为10000u/ml-300000u/ml,酶稳定剂含量为4%-10%(w/v),表面活性剂含量为2%-12%(v/v)。
优选地,所述洗涤剂为液体浓缩洗涤剂。
优选地,所述低温蛋白酶为嗜碱芽孢杆菌(Bacillus alcalophilus)发酵产生的低温蛋白酶。
优选地,所述表面活性剂为月桂醇聚氧乙烯醚、异构十醇聚氧乙烯醚、或烷基酚聚氧乙烯醚。
优选地,所述酶稳定剂包括:乳酸钙、1,2丙二醇、和短肽添加剂,所述短肽添加剂为FPR、QGF、或KNP。
优选地,所述酶稳定剂中包括:乳酸钙0.5-10重量份、1,2丙二醇2-10重量份、短肽添加剂0.05-0.5重量份。
更优选地,所述酶稳定剂中包括:乳酸钙1-3重量份、1,2丙二醇3-8重量份、短肽添加剂0.1-0.3重量份。
在另一优选例中,所述液体酶添加剂的酶活力优选为100000u/ml-200000u/ml。
在另一优选例中,所述表面活性剂的含量优选为5%-10%(v/v)。
在另一优选例中,所述洗涤剂的pH为6-10,优选为6.5-9.5。
本发明的浓缩的液体酶洗涤剂优选地为待加入液体洗涤剂的浓缩产品。由于液体酶添加剂中所用的低温酶量比较大,极易产生沉淀,且低温酶的酶活很不稳定。优选地实施方式中,浓缩的液体酶洗涤剂中的酶量是至少10000u/ml,并且不超过300000u/ml。
应理解,在本发明范围内中,本发明的上述各技术特征和在下文(如实施例)中具体描述的各技术特征之间都可以互相组合,从而构成新的或优选的技术方案。限于篇幅,在此不再一一累述。
具体实施方式
下面结合具体实施例,进一步阐述本发明。应理解,这些实施例仅用于说明本发明而不用于限制本发明的范围。下列实施例中未注明具体条件的实验方法,通常按照制造厂商所建议的条件。除非另外说明,否则百分比和份数按重量计算。
实施例1冷水低温洗涤剂的配制
1.短肽添加剂的设计与筛选
为达到提高液体的低温蛋白酶储存活性,提高产品的品相,我们开发了一种能够提高液体低温蛋白酶稳定性的低温蛋白酶抑制剂。
首先根据低温蛋白酶的酶切位点,参考低温蛋白酶的立体结构信息,设计了数十条短肽,代表性的短肽如表1所示,其中氨基酸残基由单字母缩写表示,所设计的短肽交由吉尔生化(上海)有限公司合成。
2.配制液体浓缩洗涤剂并检测保存效果
配制酶稳定剂,酶稳定剂中包括:乳酸钙2g、1,2丙二醇6g、和短肽添加剂0.2g,混合均匀。取100ml低温蛋白酶酶液(200000U/ml),搅拌加入上述配制的酶稳定剂,然后加入5%(v/v)月桂醇聚氧乙烯醚。
调节以上各组液体浓缩洗涤剂pH为6.5,对照组中不添加短肽添加剂其余组分一致,25℃保存2个月后,检测酶活保存率。
酶活检测方法、使用的试剂和仪器参考国标GBT23527-2009
嗜碱芽孢杆菌(Bacillus alcalophilus ATCC 21522)低温蛋白酶购自夏盛(上海)生物科技有限公司,由嗜碱芽孢杆菌(Bacillus alcalophilus ATCC 21522)发酵制备。酶活性检测委托夏盛(上海)生物科技有限公司应用技术研究开发中心进行并出具检测报告,检测方法如下:
1酶活力计算
X=A×K×8/2×1/10×n×E=2/5×A×K×n×E
式中:
X:样品的酶活力(u/g或u/ml);
A:试样溶液的平均吸光度;
K:吸光常数;
8:反应试剂的总体积(ml);
2:吸取酶液2.00ml,以ml计;
1/10:反应时间10min,以min计;
n:稀释倍数;
E:紫外法与福林法的换算系数(中性、碱性系数为0.50,酸性蛋白酶系数为0.77)所得结果表示至整数。
2酶活保存率
X=E1/E×100%
X:样品的酶活力保存率%
E1:样品实测酶活力(u/g或u/ml)
E:样品标示(保存前)酶活力(u/g或u/ml)
实验结果如表1所示。从表1中可以看出,1号短肽、2号短肽、3号短肽、7号短肽表现出了较好的酶活保存效果。
表1
短肽编号 | 短肽序列 | 酶活保存率% | 品相变化 |
1 | FPR | 75.6% | 澄清 |
2 | KNP | 66.2% | 澄清 |
3 | QGF | 74.3% | 澄清 |
4 | HIG | 26.4% | 稍浑浊 |
5 | KNY | 16.8% | 稍浑浊 |
6 | LNH | 33.6% | 澄清 |
7 | QRY | 43.1% | 澄清 |
对照 | 无 | 10.7% | 浑浊 |
表1的结果表明,1号短肽、2号短肽、3号短肽、7号短肽的效果较好。然后我们进一步考察了pH对酶活稳定性的影响,调节不同的pH,检测不同的短肽序列在各pH条件下表现出的酶活稳定性,各实验组设置三组平行,检测方法同上。
表2
表2的结果结合表1,可以看出短肽1在pH为7.5时,酶活保存效果最好;短肽2在pH为9.5时,酶活保存效果最好;而短肽3在pH为7.5时,酶活保存效果最好。
在本发明提及的所有文献都在本申请中引用作为参考,就如同每一篇文献被单独引用作为参考那样。此外应理解,在阅读了本发明的上述讲授内容之后,本领域技术人员可以对本发明作各种改动或修改,这些等价形式同样落于本申请所附权利要求书所限定的范围。
Claims (8)
1.一种冷水低温洗涤剂,其特征在于,所述冷水低温洗涤剂包括低温蛋白酶、酶稳定剂和表面活性剂;其中,所述低温蛋白酶含量为10000u/ml-300000u/ml,酶稳定剂含量为4%-10% w/v,表面活性剂含量为2%-12% v/v;所述低温蛋白酶为嗜碱芽孢杆菌(Bacillusalcalophilus)发酵产生的低温蛋白酶;
所述酶稳定剂包括:乳酸钙、1,2丙二醇、和短肽添加剂,所述短肽添加剂为FPR、QGF或KNP。
2.如权利要求1所述的冷水低温洗涤剂,其特征在于,所述短肽添加剂为FPR。
3.如权利要求1所述的冷水低温洗涤剂,其特征在于,所述洗涤剂为液体浓缩洗涤剂。
4.如权利要求1所述的冷水低温洗涤剂,其特征在于,所述短肽添加剂为QGF。
5.如权利要求4所述的冷水低温洗涤剂,其特征在于,所述酶稳定剂中包括:乳酸钙0.5-10重量份、1,2丙二醇2-10重量份、短肽添加剂0.05-0.5重量份。
6.如权利要求1所述的冷水低温洗涤剂,其特征在于,乳酸钙1-3重量份、1,2丙二醇3-8重量份、短肽添加剂0.1-0.3重量份。
7.如权利要求1所述的冷水低温洗涤剂,其特征在于,所述表面活性剂的含量为5%-10% v/v。
8.如权利要求1所述的冷水低温洗涤剂,其特征在于,所述洗涤剂的pH为6-10。
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CN1238003A (zh) * | 1996-09-24 | 1999-12-08 | 普罗格特-甘布尔公司 | 含有蛋白酶和蛋白酶抑制剂的液体洗衣洗涤剂组合物 |
CN103131555A (zh) * | 2013-02-05 | 2013-06-05 | 山西勇宁记科技有限公司 | 一种用于液体洗涤剂的生物酶制剂复合稳定剂 |
CN103897879A (zh) * | 2012-12-27 | 2014-07-02 | 青岛锦涟鑫商贸有限公司 | 一种强效洗涤剂 |
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CN1238003A (zh) * | 1996-09-24 | 1999-12-08 | 普罗格特-甘布尔公司 | 含有蛋白酶和蛋白酶抑制剂的液体洗衣洗涤剂组合物 |
CN103897879A (zh) * | 2012-12-27 | 2014-07-02 | 青岛锦涟鑫商贸有限公司 | 一种强效洗涤剂 |
CN103131555A (zh) * | 2013-02-05 | 2013-06-05 | 山西勇宁记科技有限公司 | 一种用于液体洗涤剂的生物酶制剂复合稳定剂 |
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