Ariaeenejad Shohreh, Zolfaghari Behrouz, Sadeghian Motahar Seyedeh Fatemeh, Kavousi Kaveh, Maleki Morteza, Roy Swapnoneel, Hosseini Salekdeh Ghasem
Department of Systems and Synthetic Biology, Agricultural Biotechnology Research Institute of Iran, Agricultural Research Education and Extension Organization, Karaj, Iran.
Department of Computer Science and Engineering, Indian Institute of Technology Guwahati, Guwahati, India.
Front Microbiol. 2021 Aug 30;12:713125. doi: 10.3389/fmicb.2021.713125. eCollection 2021.
α-Amylases are among the very critical enzymes used for different industrial purposes. Most α-amylases cannot accomplish the requirement of industrial conditions and easily lose their activity in harsh environments. In this study, a novel α-amylase named PersiAmy1 has been identified through the multistage screening pipeline from the rumen metagenomic data. The long-term storage of PersiAmy1 in low and high temperatures demonstrated 82.13 and 71.01% activities after 36 days of incubation at 4 and 50°C, respectively. The stable α-amylase retained 61.09% of its activity after 180 min of incubation at 90°C and was highly stable in a broad pH range, showing 60.48 and 86.05% activities at pH 4.0 and pH 9.0 after 180 min of incubation, respectively. Also, the enzyme could resist the high-salinity condition and demonstrated 88.81% activity in the presence of 5 M NaCl. PersiAmy1 showed more than 74% activity in the presence of various metal ions. The addition of the detergents, surfactants, and organic solvents did not affect the α-amylase activity considerably. Substrate spectrum analysis showed that PersiAmy1 could act on a wide array of substrates. PersiAmy1 showed high stability in inhibitors and superb activity in downstream conditions, thus useful in detergent and baking industries. Investigating the applicability in detergent formulation, PersiAmy1 showed more than 69% activity after incubation with commercial detergents at different temperatures (30-50°C) and retained more than 56% activity after incubation with commercial detergents for 3 h at 10°C. Furthermore, the results of the wash performance analysis exhibited a good stain removal at 10°C. The power of PersiAmy1 in the bread industry revealed soft, chewable crumbs with improved volume and porosity compared with control. This study highlights the intense power of robust novel PersiAmy1 as a functional bio-additive in many industrial applications.
α-淀粉酶是用于不同工业目的的非常关键的酶之一。大多数α-淀粉酶无法满足工业条件的要求，并且在恶劣环境中容易失去活性。在本研究中，通过多阶段筛选流程从瘤胃宏基因组数据中鉴定出一种名为PersiAmy1的新型α-淀粉酶。PersiAmy1在低温和高温下长期储存，分别在4°C和50°C孵育36天后，活性分别为82.13%和71.01%。这种稳定的α-淀粉酶在90°C孵育180分钟后仍保留61.09%的活性，并且在较宽的pH范围内高度稳定，在pH 4.0和pH 9.0孵育180分钟后，活性分别为60.48%和86.05%。此外，该酶能够抵抗高盐条件，在5 M NaCl存在下活性为88.81%。PersiAmy1在各种金属离子存在下表现出超过74%的活性。添加洗涤剂、表面活性剂和有机溶剂对α-淀粉酶活性影响不大。底物谱分析表明，PersiAmy1可以作用于多种底物。PersiAmy1在抑制剂中表现出高稳定性，在下游条件下表现出优异活性，因此可用于洗涤剂和烘焙行业。研究其在洗涤剂配方中的适用性时，PersiAmy1在不同温度（30 - 50°C）与商业洗涤剂孵育后活性超过69%，在10°C与商业洗涤剂孵育3小时后活性保留超过56%。此外，洗涤性能分析结果表明在10°C时污渍去除效果良好。PersiAmy1在面包行业的作用显示，与对照相比，面包屑柔软、可咀嚼，体积和孔隙率有所改善。本研究突出了强大的新型PersiAmy1作为功能性生物添加剂在许多工业应用中的强大作用。
