Department of Biotechnology, Central Leather Research Institute, Adyar, Chennai, Tamil Nadu, India.
J Basic Microbiol. 2014 Apr;54(4):285-95. doi: 10.1002/jobm.201200561. Epub 2013 Apr 2.
This paper describes the isolation of a potent extracellular urease producing microorganism, identified by 16S rRNA as Arthrobacter creatinolyticus MTCC 5604 and its medium optimization by classical one-factor-at-a-time method and central composite rotatable design (CCRD), a tool of response surface methodology (RSM). An optimal activity of 9.0 U ml(-1) was obtained by classical method and statistical optimization of the medium resulted in an activity of 17.35 U ml(-1) at 48 h and 30 °C. This activity was 4.91 times greater than the initial activity (3.53 U ml(-1) ) from the basal medium and the enzyme showed maximum activity at pH 8.0 and 60 °C and was stable at pH 7.0-9.0 and temperatures up to 50 °C. Furthermore, the enzyme was assessed for its activity reduction by determining the inhibitory concentration (IC50 ) of heavy metal ions and the inhibition of urease was in the order of Cu(II) > Cd(II) > Zn(II) > Ni(II). Urease was highly sensitive to Cu(II) and its inhibition was 94% and 100% in model solutions containing a mixture of Cu(II) with heavy metal ions Cd(II) and Zn(II), respectively. The results of these studies suggested that the enzyme could be utilized as sensors to determine the levels of Cu(II) ions in industrial effluents, contaminated soil and ground water.
本文描述了一种产脲酶的微生物的分离,该微生物通过 16S rRNA 鉴定为节杆菌属的解蛋白 Arthrobacter creatinolyticus MTCC 5604,并通过经典的单因素法和中心复合旋转设计(CCRD)对其培养基进行了优化,CCRD 是响应面法(RSM)的一种工具。通过经典方法获得了 9.0 U/ml(-1) 的最佳活性,通过统计优化培养基,在 48 h 和 30°C 时获得了 17.35 U/ml(-1) 的活性。该活性比基础培养基中的初始活性(3.53 U/ml(-1) )高 4.91 倍,该酶在 pH 8.0 和 60°C 时表现出最大活性,在 pH 7.0-9.0 和温度高达 50°C 时稳定。此外,通过测定重金属离子的抑制浓度(IC50)来评估酶的活性降低情况,发现脲酶的抑制顺序为 Cu(II) > Cd(II) > Zn(II) > Ni(II)。脲酶对 Cu(II) 高度敏感,在含有 Cu(II)与重金属离子 Cd(II)和 Zn(II)混合物的模型溶液中,其抑制率分别为 94%和 100%。这些研究结果表明,该酶可用于作为传感器来测定工业废水中、污染土壤和地下水中 Cu(II)离子的含量。