Karakus Betul Zehra, Korkmaz İlknur, Demirci Kubra, Sinan Arslan Kadir, Unlu Ozge, Catal Tunc
a Istanbul Protein Reserach and Inovation Center (PROMER) , Istanbul , Turkey.
b Department of Molecular Biology and Genetics , Uskudar University , Istanbul , Turkey.
Prep Biochem Biotechnol. 2018;48(9):815-822. doi: 10.1080/10826068.2018.1509088. Epub 2018 Sep 28.
In this study, three Bacillus sp.-producing amylase enzymes were isolated from soil samples and identified using 16S rDNA sequence analysis. Amylase production and total protein productions were spectrophotometrically measured. The following media were tested to increase enzyme production: LB medium and molasses. Three Bacillus sp. were identified as follows: Bacillus subtilis subtilis, Bacillus thuringiensis, and Bacillus cereus. Amylase production levels were in the range of 10 U/mL, whereas total protein production levels were at 15 mg/mL. Higher amylase activity was found in the Bacillus subtilis isolate. Ethylmethane sulfonate (EMS) and ultraviolet (UV) mutagenesis in combination were applied to compare amylase production. Amylase activity was increased to around 58% in the treatment with 0.03 mL of EMS and UV when compared to the control group. A pilot scale bioreactor with a total working volume of 10 liters was used to produce amylase by B. subtilis subtilis. In conclusion, B. subtilis subtilis can be used to produce amylase enzyme for various industrial purposes, and, for the first time, the amylase activities of B. subtilis can be enhanced with EMS and UV treatment.
在本研究中，从土壤样品中分离出三种产淀粉酶的芽孢杆菌属菌株，并通过16S rDNA序列分析进行鉴定。采用分光光度法测定淀粉酶产量和总蛋白产量。测试了以下培养基以提高酶产量：LB培养基和糖蜜。三种芽孢杆菌属菌株鉴定如下：枯草芽孢杆菌枯草亚种、苏云金芽孢杆菌和蜡样芽孢杆菌。淀粉酶产量水平在10 U/mL范围内，而总蛋白产量水平为15 mg/mL。在枯草芽孢杆菌分离物中发现了较高的淀粉酶活性。联合应用甲基磺酸乙酯（EMS）和紫外线（UV）诱变来比较淀粉酶产量。与对照组相比，用0.03 mL EMS和UV处理时淀粉酶活性提高到约58%。使用总工作体积为10升的中试规模生物反应器由枯草芽孢杆菌枯草亚种生产淀粉酶。总之，枯草芽孢杆菌枯草亚种可用于为各种工业目的生产淀粉酶，并且首次通过EMS和UV处理提高了枯草芽孢杆菌的淀粉酶活性。
