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链霉菌属MR28菌株酪氨酸酶的生物生产、纯化、部分特性表征及苯酚去除效果

Bioproduction, purification, partial characterization and phenol removal efficacy of tyrosinase enzyme from Streptomyces sp. strain MR28.

作者信息

Rudrappa Muthuraj, Santosh Kumar M, Basavarajappa Dhanyakumara Shivapoojar, Hiremath Halaswamy, Hugar Anil, Almansour Abdulrahman I, Kantli Gireesh Babu, Nayaka Sreenivasa

机构信息

P.G. Department of Studies in Botany, Karnatak University, Dharwad, 580003, Karnataka, India.

Department of Studies in Biochemistry, Davangere University, Davangere, 577007, Karnataka, India.

出版信息

Environ Res. 2024 Jun 15;251(Pt 2):118701. doi: 10.1016/j.envres.2024.118701. Epub 2024 Mar 18.

Abstract

The study focused on the production of the tyrosinase enzyme from Streptomyces sp. MR28 and its potency in removal of phenol content from water using free and immobilized tyrosinase enzyme. The tyrosinase was produced by Streptomyces sp. MR28 in liquid tyrosine broth medium, and it was further purified to near its homogeneity by employing, precipitation, dialysis, and column chromatography. After the purification, 44.49% yield with a 4 fold purification was achieved. The characterization of the purified enzyme showed a single major peak on HPLC and a solitary band on SDS-PAGE. The purified tyrosinase enzyme was active at a pH of 7.0 and a temperature of 30 °C. Further immobilization of purified tyrosinase was performed using the sodium alginate entrapment method. The capacity of the purified tyrosinase to remove phenol in water was evaluated by spectrophotometric method. The free tyrosinase enzyme-treated solutions showed a gradual decrease in the concentration of phenol with increased incubation time at 30 °C and 40 °C, at 90 min of the incubation time, it showed maximum efficacy in removing phenol from the solution. At 50 °C and 60 °C, the free tyrosinase enzyme exhibited very less capacity to remove the phenol. The immobilized enzyme showed good capacity for the removal of phenol from the solutions; the concentration of phenol in the solution decreased with an increase in the incubation time. At temperatures of 40 °C and 50 °C, the immobilized tyrosinase enzyme beads showed significant removal of phenol from the solution, and at temperatures of 30 °C and 60 °C, they also exhibited good capacity for the removal of phenol. At the end of the 90 min incubation period, it exhibited good capability. The current study suggests using immobilized microbial tyrosinase enzyme can be used for the removal of phenol from the contaminated water in a greener manner.

摘要

该研究聚焦于链霉菌属MR28产酪氨酸酶及其使用游离和固定化酪氨酸酶从水中去除苯酚含量的效能。酪氨酸酶由链霉菌属MR28在液体酪氨酸肉汤培养基中产生,并通过沉淀、透析和柱色谱法进一步纯化至接近均一性。纯化后,获得了44.49%的产率和4倍的纯化倍数。纯化酶的表征显示在高效液相色谱上有一个主要主峰,在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳上有一条单独条带。纯化的酪氨酸酶在pH 7.0和温度30℃时具有活性。使用海藻酸钠包埋法对纯化的酪氨酸酶进行进一步固定化。通过分光光度法评估纯化的酪氨酸酶去除水中苯酚的能力。游离酪氨酸酶处理的溶液在30℃和40℃下随着孵育时间的增加苯酚浓度逐渐降低,在孵育90分钟时,其在从溶液中去除苯酚方面显示出最大功效。在50℃和60℃时,游离酪氨酸酶去除苯酚的能力非常低。固定化酶显示出从溶液中去除苯酚的良好能力;溶液中苯酚的浓度随着孵育时间的增加而降低。在40℃和50℃时,固定化酪氨酸酶珠显示出从溶液中显著去除苯酚,在30℃和60℃时,它们也表现出良好的去除苯酚的能力。在90分钟孵育期结束时,它表现出良好的能力。当前研究表明,使用固定化微生物酪氨酸酶可以更环保地从受污染水中去除苯酚。

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