当前位置：
X-MOL 学术
›
ACS Sustain. Chem. Eng.
›
论文详情
Our official English website, www.x-mol.net, welcomes your
feedback! (Note: you will need to create a separate account there.)
Modifying the Substrate Specificity of Keratinase for Industrial Dehairing to Replace Lime-Sulfide
ACS Sustainable Chemistry & Engineering ( IF 7.1 ) Pub Date : 2022-05-13 , DOI: 10.1021/acssuschemeng.2c02352 Zheng Peng 1, 2 , Xinzhe Mao 1 , Wanmeng Mu 3 , Guocheng Du 1, 2 , Jian Chen 1, 2 , Juan Zhang 1, 2
ACS Sustainable Chemistry & Engineering ( IF 7.1 ) Pub Date : 2022-05-13 , DOI: 10.1021/acssuschemeng.2c02352 Zheng Peng 1, 2 , Xinzhe Mao 1 , Wanmeng Mu 3 , Guocheng Du 1, 2 , Jian Chen 1, 2 , Juan Zhang 1, 2
Affiliation
|
Keratinase has shown potential as an ecofriendly alternative to toxic lime-sulfide for industrial dehairing. However, low substrate specificity usually causes collagen damage. Here, we developed a variant of keratinase KerZ1 with low collagenase activity by modifying the substrate binding pocket. First, sequence alignment identified several targets for distinguishing substrate selectivity. Although the keratinase activity decreased by 41.16 and 22.99%, the collagenase activity of variants Y208V and L216S decreased by 70.91 and 49.51%. On the other hand, molecular docking of KerZ1 with the characteristic peptide FALGPA also revealed that the keratinase activity of variant S100D increased by 5.45%, and the collagenase activity decreased by 25.90%. Next, the keratinase activity of the double mutagenesis variant S100D/Y208V was only 0.04% lower than that of KerZ1, while collagenase activity greatly reduced by 60%. Moreover, the catalytic efficiency (Kcat/Km) of the variant S100D/Y208V on collagen decreased by 79.60%. For dehairing, S100D/Y208V not only significantly reduced the damage to cowhide and sheepskin collagen but also maintained the same dehairing ability as KerZ1. In fed-batch fermentation, the keratinase activity of S100D/Y208V increased by 9.28 times. This study suggests that improving the substrate specificity will promote keratinase to replace toxic lime-sulfide for industrial dehairing.
中文翻译：
修改工业脱毛用角蛋白酶的底物特异性以替代硫化石灰
角蛋白酶已显示出作为有毒硫化石灰的环保替代品用于工业脱毛的潜力。然而，低底物特异性通常会导致胶原蛋白损伤。在这里，我们通过修改底物结合口袋开发了一种具有低胶原酶活性的角蛋白酶 KerZ1 变体。首先，序列比对确定了几个用于区分底物选择性的目标。虽然角蛋白酶活性下降了 41.16% 和 22.99%，但变体 Y208V 和 L216S 的胶原酶活性下降了 70.91% 和 49.51%。另一方面，KerZ1与特征肽FALGPA的分子对接也显示变体S100D的角蛋白酶活性增加了5.45%，胶原酶活性降低了25.90%。其次，双诱变变体S100D/Y208V的角蛋白酶活性仅为0。比KerZ1低04%，而胶原酶活性大大降低60%。此外，催化效率（K cat / K m ) 变体 S100D/Y208V 对胶原蛋白的影响降低了 79.60%。在脱毛方面，S100D/Y208V不仅显着降低了对牛皮和羊皮胶原蛋白的损伤，而且保持了与KerZ1相同的脱毛能力。在补料分批发酵中，S100D/Y208V的角蛋白酶活性提高了9.28倍。该研究表明，提高底物特异性将促进角蛋白酶替代有毒的硫化石灰用于工业脱毛。
更新日期：2022-05-13
中文翻译：
修改工业脱毛用角蛋白酶的底物特异性以替代硫化石灰
角蛋白酶已显示出作为有毒硫化石灰的环保替代品用于工业脱毛的潜力。然而，低底物特异性通常会导致胶原蛋白损伤。在这里，我们通过修改底物结合口袋开发了一种具有低胶原酶活性的角蛋白酶 KerZ1 变体。首先，序列比对确定了几个用于区分底物选择性的目标。虽然角蛋白酶活性下降了 41.16% 和 22.99%，但变体 Y208V 和 L216S 的胶原酶活性下降了 70.91% 和 49.51%。另一方面，KerZ1与特征肽FALGPA的分子对接也显示变体S100D的角蛋白酶活性增加了5.45%，胶原酶活性降低了25.90%。其次，双诱变变体S100D/Y208V的角蛋白酶活性仅为0。比KerZ1低04%，而胶原酶活性大大降低60%。此外，催化效率（K cat / K m ) 变体 S100D/Y208V 对胶原蛋白的影响降低了 79.60%。在脱毛方面，S100D/Y208V不仅显着降低了对牛皮和羊皮胶原蛋白的损伤，而且保持了与KerZ1相同的脱毛能力。在补料分批发酵中，S100D/Y208V的角蛋白酶活性提高了9.28倍。该研究表明，提高底物特异性将促进角蛋白酶替代有毒的硫化石灰用于工业脱毛。
京公网安备 11010802027423号