Isolation and characterization of tyrosinase produced by marine actinobacteria and its application in the removal of phenol from aqueous environment

doi:10.1007/s11515-014-1324-0

Front. Biol.

2014, Vol. 9

Issue (4) : 306-316 https://doi.org/10.1007/s11515-014-1324-0

RESEARCH ARTICLE

Isolation and characterization of tyrosinase produced by marine actinobacteria and its application in the removal of phenol from aqueous environment

Suki ROY¹,Ishita DAS¹,Minki MUNJAL¹,Loganathan KARTHIK^1,²,Gaurav KUMAR^1,²,Sathish KUMAR¹,Kokati Venkata Bhaskara RAO^1,^*(

)

1. Molecular and Microbiology Research Laboratory, School of Bio Sciences and Technology, VIT University, Vellore-632014, Tamil Nadu, India
2. Department of Biotechnology, Shri JJT University, Jhunjhunu, Rajasthan, India

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Abstract

The present study was focused on screening and characterization of tyrosinase enzyme produced by marine actinobacteria and its application in phenolic compounds removal from aqueous solution. A total of 20 strains were isolated from marine sediment sample and screened for tyrosinase production by using skimmed milk agar medium. Among 20 isolates, two isolates LK-4 and LK-20 showed zone of hydrolysis and these were taken for secondary screening by using tyrosine agar medium. Based on the result of secondary screening LK-4 was selected for further analysis, such as tyrosinase assay, protein content and specific activity of the enzyme. The tyrosinase enzyme was produced in a SS medium and was partially purified by ammonium sulfate precipitation, dialysis and SDS PAGE. The isolate (LK-4) was identified as Streptomyces espinosus using 16S rRNA gene sequencing and named as “Streptomyces espinosus strain LK4 (KF806735)”. The tyrosinase enzyme was immobilized in sodium alginate which was applied to remove phenolic compounds from water. The enzyme efficiently removed the phenolic compounds from aqueous solution within few hours which indicated that tyrosinase enzyme produced by Streptomyces espinosus strain LK-4 can be potently used for the removal of phenol and phenolic compounds from wastewater in industries.

Keywords actinobacteria phenolic compounds immobilization tyrosinase

Corresponding Author(s): Kokati Venkata Bhaskara RAO

Issue Date: 11 August 2014

Cite this article:

Suki ROY,Ishita DAS,Minki MUNJAL, et al. Isolation and characterization of tyrosinase produced by marine actinobacteria and its application in the removal of phenol from aqueous environment[J]. Front. Biol., 2014, 9(4): 306-316.

URL:

https://academic.hep.com.cn/fib/EN/10.1007/s11515-014-1324-0
https://academic.hep.com.cn/fib/EN/Y2014/V9/I4/306

Fig.1 Secondary screening of tyrosinase production by LK-4 and LK-20. (A) Tyrosine agar plate, (B) tyrosine broth.

Fig.2 SDS-PAGE of tyrosinase produced by LK-4.

Fig.3 HPLC profile of partially purified tyrosinase enzyme from LK-4.

Fig.4 Optimum pH for tyrosinase production by LK-4. Data is given as mean±SD (n=3).

Fig.5 Optimum temperature for tyrosinase production by LK-4. Data is given as mean ± SD (n =3).

Fig.6 Optimum concentration of tyrosine for tyrosinase production by LK-4. Data is given as mean ± SD (n =3).

Fig.7 Phylogenetic tree of Streptomyces espinosus strain LK-4 (KF806735).

Fig.8 Removal of different concentrations of phenol from aqueous solution by immobilized tyrosinase enzyme produced by Streptomyces espinosus strain LK-4.(A) Phenol 1mM. (B) Phenol 2 mM. (C) Phenol 3 mM. (D) Phenol 4 mM. (E) Phenol 5 mM.

Tab.1 Secondary screening of tyrosinase produced by two isolate LK-4 and LK-20

Tab.2 Characterization of tyrosinase produced by isolate LK-4

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