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Production, purification and applications of raw starch degrading and calcium-independent α-amylase from soil rich in extremophile.
International Journal of Biological Macromolecules ( IF 7.7 ) Pub Date : 2020-06-19 , DOI: 10.1016/j.ijbiomac.2020.06.160
Isha Kohli 1 , Naveen Chandra Joshi 1 , Ajit Varma 1
Affiliation  

Calcium independent, raw starch hydrolyzing, acidic α-amylase (66 kDa) was synthesized by Bacillus subtilis S113 that is an aerobic, rod-shaped and Gram +ve bacteria. Purification of the enzyme was performed by HiTrap Capto Q (Ion-exchange chromatography; 19.28 fold; 22.41% yield). The purified enzyme was found stable at broad acidic pH (4–6.5) and high-temperature range (40–80 °C), that fulfilled the necessary criteria and laid the foundation to be utilized in starch saccharification industry. Kinetic studies of the enzyme revealed that Km and Vmax of the enzyme was 0.22% and 357.14 U/mg respectively. Scanning electron microscopy studies showed that the enzyme was capable of completely hydrolyzing raw wheat and potato starch, further confirming its role in the starch industry. It was found that only 7.93% of the activity was loss at 4 °C when kept for one year.



中文翻译:

从富含极端微生物的土壤中降解淀粉和不依赖钙的原始淀粉的生产,纯化和应用。

枯草芽孢杆菌S113合成了不依赖钙的粗淀粉水解酸性α-淀粉酶(66 kDa),它是一种好氧的杆状革兰氏+细菌。通过HiTrap Capto Q(离子交换色谱; 19.28倍; 22.41%收率)进行酶的纯化。发现纯化的酶在宽的酸性pH(4–6.5)和高温范围(40–80°C)下稳定,满足了必要的标准,并为淀粉糖化工业中的应用奠定了基础。酶的动力学研究表明,K m和V max酶的分别为0.22%和357.14U / mg。扫描电子显微镜研究表明,该酶能够完全水解生小麦和马铃薯淀粉,进一步证实了其在淀粉工业中的作用。发现在保存一年后,在4°C下仅损失7.93%的活性。

更新日期:2020-07-02
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