摘要
酶制剂是洗衣液的重要组分,在这种环境中蛋白酶不仅会自溶,也会水解其他酶类。通过添加蛋白酶稳定剂可以暂时抑制蛋白酶的活性,提高洗衣液中酶的稳定性。现已成功商业化的两种稳定剂受专利保护,无法广泛应用,因此开发更多高效的稳定剂对洗涤酶的发展非常重要。常规的稳定剂的测试验证耗时长,效率低,文章基于稳定剂本质为可逆性抑制剂的原理,开发了简便高效的筛选方法。首先利用蛋白酶水解对硝基苯酚乙酸酯(p-NPA)反应可定量检测出候选物质对蛋白酶的抑制效率,再验证它们在洗涤剂中对酶的稳定能力,其中以脂肪酶残留率为指标可进一步缩短开发周期。利用这种方法对多种物质进行了比较,发现所测已报导的稳定剂中2,6-二氟吡啶-3-硼酸效果最好,另外发现4-(烷基磺酰基)苯硼酸系列物质也可做蛋白酶稳定剂。使用本方法测定的蛋白酶抑制率与含蛋白酶洗涤剂中的脂肪酶残余率有中度线性关系,且不受所测试蛋白酶种类影响。结果表明本方法适合高通量筛选蛋白酶稳定剂。
Enzyme is a kind of important ingredient of most liquid laundry detergent.Protease will autolyze and hydrolyze other enzymes in the liquid.By adding protease stabilizers,the activity of protease is inhibited temporarily,and the shelf life will be prolonged.There are already two commercialized stabilizers in the market,but they could not be used widely due to patent protection.Therefore,to develop other efficient stabilizers is very important for the users.The conventional verification test of stabilizers takes a very long time,so it is not suitable for high-throughput screening.Here this paper developed a simple and efficient screening method based on the principle that stabilizer is a reversible inhibitor.Firstly,the protease hydrolysis reaction of p-nitrophenyl acetate(p-NPA)was used to quantify the candidates’inhibition ratio on protease,and then verified their stability efficiency in detergents.In addition,lipase can be used as an indicator to further shorten the experiment period.By this way,a variety of substances were evaluated and compared.As a result,2,6-difluoropyridine-3-boronic acid is found to be the most efficient one among the reported stabilizers,and 4-(alkylsulfonyl)phenylboronic acid may be also suitable as stabilizers.Finally,there is a positive moderate linear correlation between the protease inhibition ratio and the lipase residual rate in the protease-containing detergents,independent of protease species.The results indicate that this method is suitable for high-throughput screening of protease stabilizers and may be helpful for the development of detergent enzymes.
作者
王雪娇
杜丽娟
徐元喜
Xuejiao Wang;Lijuan Du;Yuanxi Xu(Nanjing Bestzyme Bio-Engineering Co.,Nanjing,Jiangsu 211100,China)
出处
《日用化学工业（中英文）》
CAS
北大核心
2023年第7期796-801,共6页
China Surfactant Detergent & Cosmetics
关键词
洗涤酶
蛋白酶
稳定剂
抑制剂
detergent enzymes
protease
stabilizer
inhibitor
