Marathe Sarika Kedar, Vashistht Manisha Arun, Prashanth Aishwarya, Parveen Nikhat, Chakraborty Shailayee, Nair Sindhu S
Department of Biotechnology, Mount Carmel College, Bengaluru, Karnataka, India.
J Genet Eng Biotechnol. 2018 Jun;16(1):39-46. doi: 10.1016/j.jgeb.2017.10.001. Epub 2017 Oct 12.
In the current study, bacteria isolated from sea water samples of Murdeshwar, Karnataka, were screened for the production of alkaline protease by culturing them onto skim milk agar media. Of the isolated bacteria, , and showed distinct zones of hydrolysis due to enzyme production. They were each inoculated into enzyme production media under submerged fermentation conditions at 37 °C for 48 h with a constant agitation of 120 rpm. Partial purification of alkaline protease was carried out by isoelectric precipitation. Enzyme activity was determined under varying conditions of pH, incubation temperature, different substrates, carbon and nitrogen sources and salt concentrations using sigma's universal protease activity assay. Enzyme immobilization was carried out using 2% Sodium alginate and 0.1 M ice cold CaCl and its activity under varying pH, temperature conditions and detergent compatibility was assayed. Efficacy of enzyme in stain removal was tested and haemolysis was observed within of 60 s which resulted in removal of the stain. Among the three organisms, enzyme from showed highest activity in all cases indicating that it was the most ideal organism for enzyme production.
在当前研究中，从卡纳塔克邦穆尔代什瓦尔海水样本中分离出的细菌，通过在脱脂乳琼脂培养基上培养来筛选碱性蛋白酶的产生情况。在分离出的细菌中，[此处原文缺失具体细菌名称]、[此处原文缺失具体细菌名称]和[此处原文缺失具体细菌名称]由于产酶而显示出明显的水解圈。将它们分别接种到产酶培养基中，在37℃的深层发酵条件下培养48小时，持续搅拌速度为120转/分钟。通过等电沉淀法对碱性蛋白酶进行部分纯化。使用西格玛通用蛋白酶活性测定法，在不同的pH值、孵育温度、不同底物、碳源和氮源以及盐浓度条件下测定酶活性。使用2%海藻酸钠和0.1M冰冷氯化钙进行酶固定化，并测定其在不同pH值、温度条件下以及与洗涤剂的兼容性下的活性。测试了酶在去除污渍方面的功效，观察到在60秒内出现溶血现象，从而导致污渍去除。在这三种微生物中，[此处原文缺失具体细菌名称]产生的酶在所有情况下都表现出最高活性，表明它是产酶的最理想微生物。
