Screening, Isolation and Identification of Thermophilic Esterase Enzyme Isolated from Rhodococcus SP: LKE-021

Singh, L. N.; Sharma, Gaurav; Awasthi, Gyanendra; Kumar, Lalit; Ali, Mohammad; Moin, Sarmad

doi:10.22207/jpam.13.3.63

Cited by 5 publications

(2 citation statements)

References 24 publications

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“…LKE-021 was inoculated in shake flasks method containing modified nutrient broth (pH 7.0) having5.0g/l NaCl; 5.0g/l peptone; 3.0g/l yeast extract, and 10.0g/l glucose with constant shaking at 135 rpm at 60°. After 24 hrs extracellular enzyme was recovered by centrifugation for 5 min at 10×10 3 relative centrifugal force in liquid fraction 15 .…”

Section: Separation Of Esterasementioning

confidence: 99%

Purification, Isolation, and Characterization of Esterase from Rhodococcus sp. LKE-021

Singh¹,

Sharma²,

Sharma³

et al. 2020

J Pure Appl Microbiol

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A thermophilic esterase isolated from Rhodococcus sp. LKE-021. This enzyme was purified with purification fold 60 from the crude extracts of enzyme and recovery of enzyme obtained approximately 21%. The specific activity of the lKe-021 esteraseis 795.1 U/mg. SDS-PAGE analysis determined the molecular weight of LKE-21 esteraseis around 32,000Da/32KDa. The enzyme activity of lKe-021 esterase exhibited over a wide range of temperature i.e. 30° to 80°C and the enzyme remained stable when incubated on 60° for 2h. This indicates that the isolated lKe-021 esterase is thermostable. The isolated enzyme exhibits activity on various pH ranges from 2.0 to 12.0 and the highest activity observed on 11.0 pH.The LKE-021 esterase was active after proteinase K treatment and shows over 75 % specific activity i.e. 50 U/µg Proteinase K.

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Section: Separation Of Esterasementioning

confidence: 99%

Purification, Isolation, and Characterization of Esterase from Rhodococcus sp. LKE-021

Singh¹,

Sharma²,

Sharma³

et al. 2020

J Pure Appl Microbiol

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“…In pharmacology, esterases are used for the creation of crucial chiral chemicals owing to their great region stereospecificity [ 5 , 6 ]. The widespread use of this enzyme in different sectors of industry has sparked interest in the discovery and characterization of new esterases of microbial origin [ 7 , 8 ]. Esterases from microorganisms are potent biocatalysts that efficiently hydrolyze many types of polyester, including polylactic acid (PLA), in the process of plastic depolymerization.…”

Section: Introductionmentioning

confidence: 99%

A scalable overexpression of a thermostable recombinant poly-histidine tag carboxyl esterase under lambda promoter: purification, characterization, and protein modelling

Soliman,

Ali,

Duab

et al. 2023

Journal of Genetic Engineering and Biotechnology

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Background As a white biotechnological trend, esterases are thought to be among the most active enzymes’ classes in biocatalysis and synthesis of industrially importance organic compounds. Esterases are used in many applications such as the manufacture of pharmaceuticals, cosmetics, leather, textile, paper, food, dairy products, detergents, and treatment of some environmental pollutants. Results A poly-histidine moiety was added to the C-terminal end of the Geobacillus sp. gene encoding carboxyl esterase (EstB, ac: KJ735452) to facilitate one-step purification. This recombinant protein was successfully expressed in Escherichia coli (E. coli) under control of Lambda promoter (λ). An open reading frame (ORF) of 1500 bps encoding a polypeptide of 499 amino acid residues and a calculated molecular weight (54.7 kD) was identified as carboxyl-esterase B due to its conserved glycine-X-serine-X-glycine motif (G-X-S-X-G) and its high similarity toward other carboxyl esterases, where the 3-D tertiary structure of EstB was determined based on high homology % (94.8) to Est55. The expression was scaled up using 7-L stirred tank bioreactor, where a maximum yield of enzyme was obtained after 3.5 h with SEA 51.76 U/mg protein. The expressed protein was purified until unity using immobilized metal affinity chromatography (IMAC) charged with cobalt and then characterized. The purified enzyme was most active at pH 8.0 and remarkably stable at pH (8–10). Temperature optimum was recorded at 65 °C, and it kept 70% of its activity after 1-h exposure to 60 °C. The active half-live of enzyme was 25 min at 70 °C and a calculated T melting (Tm) at 70 °C. The determined reaction kinetics Michaelis–Menten constant (Km), maximum velocity rate (Vmax), the turnover number (Kcat), and catalytic efficiency (Kcat/Km) of the pure enzyme were found 22.756 mM, 164.47 U/ml (59.6 min−1), and (2.619 mol/ min), respectively. Conclusion Creation of a recombinant 6 × -His estB derived from a thermophile Geobacillus sp. was performed successfully and then overexpressed under λ-promoter. In a bench scale bioreactor, the overexpression was grown up, followed by one-step purification and biochemical characterization. The recorded promising pH and temperature stability properties suggest that this expressed carboxyl esterase could be used in many industrial sectors.

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The Biocatalytic Potential of Extremozymes Isolated from Extremophiles

Nivedita,

Behera,

Nayak

et al. 2024

Trends in Biotechnology of Polyextremophiles

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Screening, Isolation and Identification of Thermophilic Esterase Enzyme Isolated from Rhodococcus SP: LKE-021

Cited by 5 publications

References 24 publications

Purification, Isolation, and Characterization of Esterase from Rhodococcus sp. LKE-021

Purification, Isolation, and Characterization of Esterase from Rhodococcus sp. LKE-021

A scalable overexpression of a thermostable recombinant poly-histidine tag carboxyl esterase under lambda promoter: purification, characterization, and protein modelling

The Biocatalytic Potential of Extremozymes Isolated from Extremophiles

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