Center of Innovative and Applied Bioprocessing (CIAB), Sector-81 (Knowledge City), Mohali, 140306, PB, India.
Center of Innovative and Applied Bioprocessing (CIAB), Sector-81 (Knowledge City), Mohali, 140306, PB, India.
Colloids Surf B Biointerfaces. 2021 Aug;204:111832. doi: 10.1016/j.colsurfb.2021.111832. Epub 2021 May 9.
Here, a chimeric bifunctional enzyme was developed for two activities xylanase and deacetylase. Chimeric enzyme was designed by combining the relevant amino acid stretches from two different parent sequences, such as polysaccharide/xylan deacetylase (ref id: MT682066) and xylanase (ref id WP_110897546.1). Five different hypothetical chimeras were developed and one of the best predicted chimeric protein GA_2(syn_SKYAP01) was synthesized. The GA_2(syn_SKYAP01) possessed the specific activity of 14.905 ± 0.8 U/mg for deacetylase and 100.87 ± 14.2 U/mg for xylanase. Optimum level of both the activities together was achieved at pH 5 and 60 °C. The chimeric protein was also found to be stable at higher temperature of 71°C. Functionality of the developed chimeric protein for both the activities was confirmed by the hydrolysis of commercial xylan into xylooligosaccharides and the release of acetic acid from glucose pentacetate and 7-amino cephalosporin. The designed bifunctional enzyme was found to be highly efficient.
在这里，开发了一种具有两种活性（木聚糖酶和脱乙酰酶）的嵌合双功能酶。嵌合酶是通过组合两个不同亲本序列的相关氨基酸序列设计的，例如多糖/木聚糖脱乙酰酶（参考 ID：MT682066）和木聚糖酶（参考 ID WP_110897546.1）。开发了五种不同的假设嵌合体，其中一种最佳预测的嵌合蛋白 GA_2（syn_SKYAP01）被合成。GA_2（syn_SKYAP01）的脱乙酰酶比活为 14.905±0.8 U/mg，木聚糖酶比活为 100.87±14.2 U/mg。两种活性的最佳水平均在 pH5 和 60°C 时达到。该嵌合蛋白在 71°C 的较高温度下也表现出稳定性。通过将商业木聚糖水解成木二糖寡糖以及从葡萄糖五乙酸酯和 7-氨基头孢菌素中释放乙酸，证实了所开发的嵌合蛋白在这两种活性方面的功能。设计的双功能酶被发现具有很高的效率。
