Ameri Atefeh, Shakibaie Mojtaba, Soleimani-Kermani Mozhde, Faramarzi Mohammad Ali, Forootanfar Hamid
a Pharmaceutics Research Center, Institute of Neuropharmacology , Kerman University of Medical Sciences , Kerman , Iran.
b Department of Pharmaceutical Biotechnology, Faculty of Pharmacy , Kerman University of Medical Sciences , Kerman , Iran.
Prep Biochem Biotechnol. 2019;49(2):184-191. doi: 10.1080/10826068.2019.1566148. Epub 2019 Feb 3.
Microbial enzymes of extremophilic origin serve as a vital source of stable industrial enzymes. The present study focused on overproduction of a thermoalkalophilic lipase produced by Bacillus atrophaeus FSHM2 through UV-induced random mutagenesis (5-45 min exposure to UV light) and factorial experimental design augmented to response surface methodology. Firstly, a UV-induced mutant (designated as UV-45) was developed after the exposure of wild strain to UV irradiation for 45 min which was able to secrete 3484.8 U/L lipase. Afterward, Plackett-Burman experimental approach augmented to central composite design was employed to optimize medium components (olive oil, maltose, glucose, sucrose, yeast extract, tryptone, urea, (NH)SO, NaCl, CaCl, and ZnSO) for lipase production by the UV-45 mutant strain. The maximum lipase production of 5505.3 U/L were predicted in medium containing 5% of olive oil, 0.69% of glucose, 0.69% of sucrose, 2.5% of maltose, yeast extract (0.7 g/L), urea (0.44 g/L), (NH)SO (2.44 g/L), tryptone (1.19 g/L), NaCl (1.61 g/L), CaCl (3.81 g/L), and ZnSO (1.42 g/L). A mean value of 5161.3 ± 83.3 U/L of lipolytic activity was acquired from real experiments. To sum up, the lipolytic activity of wild type strain (1720.4 U/L) increased by 3-fold after UV-induced mutagenesis and medium components optimization (5161.3 U/L).
极端微生物来源的微生物酶是稳定的工业酶的重要来源。本研究聚焦于通过紫外线诱导的随机诱变(紫外线照射5 - 45分钟)和响应面法增强的析因实验设计,过量生产萎缩芽孢杆菌FSHM2产生的嗜热嗜碱脂肪酶。首先,野生菌株经45分钟紫外线照射后获得了一个紫外线诱导突变体(命名为UV - 45),该突变体能够分泌3484.8 U/L的脂肪酶。随后,采用Plackett - Burman实验方法并结合中心复合设计,对UV - 45突变菌株生产脂肪酶的培养基成分(橄榄油、麦芽糖、葡萄糖、蔗糖、酵母提取物、胰蛋白胨、尿素、(NH₄)₂SO₄、NaCl、CaCl₂和ZnSO₄)进行优化。在含有5%橄榄油、0.69%葡萄糖、0.69%蔗糖、2.5%麦芽糖、酵母提取物(0.7 g/L)、尿素(0.44 g/L)、(NH₄)₂SO₄(2.44 g/L)、胰蛋白胨(1.19 g/L)、NaCl(1.61 g/L)、CaCl₂(3.81 g/L)和ZnSO₄(1.42 g/L)的培养基中,预测脂肪酶产量最高可达5505.3 U/L。实际实验获得的脂肪酶活性均值为5161.3 ± 83.3 U/L。综上所述,野生型菌株(1720.4 U/L)的脂肪酶活性在紫外线诱变和培养基成分优化后提高了3倍(5161.3 U/L)。