Catalytic, thermodynamic and structural properties of an immobilized and highly thermostable alkaline protease from a haloalkaliphilic actinobacteria, Nocardiopsis alba TATA-5

Thakrar, Foram J.; Singh, Satya P.

doi:10.1016/j.biortech.2019.01.058

Cited by 50 publications

(18 citation statements)

References 37 publications

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“…The activation energy is the energy difference between the transition state and the ground state of the reactant, which determines the reaction rate (Thakrar & Singh, 2019). The activities of S. carnosus RT6 protease were assayed in a temperature range of 20 to 50 • C. Arrhenius plots of S. carnosus RT6 protease (Figure 3b) indicated that the calculated activation energy (E a ) was about 24.62 kJ/mol (…”

Section: Determination Of Activation Energymentioning

confidence: 99%

Biochemical properties of extracellular protease from Staphylococcus carnosus RT6 isolated from Harbin dry sausages, and its hydrolysis of meat proteins

Wang

Xia

et al. 2021

Journal of Food Science

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The characteristics of the extracellular protease, produced by Staphylococcus carnosus RT6 isolated from Harbin dry sausages, and its hydrolysis of meat proteins were investigated. The protease was purified by ammonium sulfate, ion exchange, and gel filtration chromatography to obtain a 20.0 kDa extracellular protease. The protease reached maximal activity at pH 9.0 and 50 °C and was stable at pH 7.0 to 11.0 and 20 to 40 °C. Its protease activity was easily inhibited in the presence of Zn2+, Fe2+, and Fe3+. The enzymatic characterization of the protease revealed a Vmax 49.50 U/ml·min, Km 8.19 mg/ml, and the half‐life = 28.06 min, ΔH*d = 114.11 kJ/mol, ΔG*d = 89.24 kJ/mol, and ΔS*d = 77.00 J/mol·K at 50 °C. In addition, the protease hydrolyzed meat protein into small particles and produced soluble peptides. This study provides a basis for understanding the biochemical characteristics of the S. carnosus RT6 protease and its future application for fermented meat products.

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Section: Determination Of Activation Energymentioning

confidence: 99%

Biochemical properties of extracellular protease from Staphylococcus carnosus RT6 isolated from Harbin dry sausages, and its hydrolysis of meat proteins

Wang

Xia

et al. 2021

Journal of Food Science

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“…Therefore, the search for suitable enzymes continues for various applications. Alkaline proteases are reported from bacteria and actinomycetes of different marine habitats (Thumar and Singh, 2009;Singh, 2012, 2018;Annamalai et al, 2014;Raval et al, 2014Raval et al, , 2015Thakrar and Singh, 2019;Sharma et al, 2020). However, only limited studies are available on the microorganisms of desert origin.…”

Section: Introductionmentioning

confidence: 99%

Cloning, Expression, and Structural Elucidation of a Biotechnologically Potential Alkaline Serine Protease From a Newly Isolated Haloalkaliphilic Bacillus lehensis JO-26

Bhatt

Singh

2020

Front. Microbiol.

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An alkaline protease gene of Bacillus lehensis JO-26 from saline desert, Little Rann of Kutch, was cloned and expressed in Escherichia coli BL21 (DE3). A 1,014-bp ORF encoded 337 amino acids. The recombinant protease (APrBL) with Asp 97, His 127, and Ser 280 forming catalytic triad belongs to the subtilase S8 protease family. The gene was optimally expressed in soluble fraction with 0.2 mM isopropyl β-D-thiogalactopyranoside (IPTG), 2% (w/v) NaCl at 28 • C. APrBL, a monomer with a molecular mass of 34.6 kDa was active over pH 8-11 and 30 • C−70 • C, optimally at pH 10 and 50 • C. The enzyme was highly thermostable and retained 73% of the residual activity at 80 • C up to 3 h. It was significantly stimulated by sodium dodecyl sulfate (SDS), Ca 2+ , chloroform, toluene, n-butanol, and benzene while completely inhibited by phenylmethylsulfonyl fluoride (PMSF) and Hg 2+ . The serine nature of the protease was confirmed by its strong inhibition by PMSF. The APrBL gene was phylogenetically close to alkaline elastase YaB (P20724) and was distinct from the well-known commercial proteases subtilisin Carlsberg (CAB56500) and subtilisin BPN (P00782). The structural elucidation revealed 31.75% α-helices, 22.55% β-strands, and 45.70% coils. Although high glycine and fewer proline residues are a characteristic feature of the cold-adapted enzymes, the similar observation in thermally active APrBL suggests that this feature cannot be solely responsible for thermo/cold adaptation. The APrBL protease was highly effective as a detergent additive and in whey protein hydrolysis.

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“…Deactivation rate constant (k d ) was determined according to Eq. ( 7) from the semi logarithmic plot of residual activity (%) versus time (min) (Singh et al 2019).…”

Section: Optimum Temperature and Thermal Stabilitymentioning

confidence: 99%

Kinetic, Catalytic and Thermodynamic Properties of Immobilized Milk Clotting Enzyme on Activated Chitosan Polymer and its Application in Cheese Making

Ahmed

Abdel–Naby

Abdelfattah³

2021

Preprint

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Milk clotting enzyme (MCE) from Bacillus circulans 25 was immobilized by covalent binding, ionic binding and entrapment methods using various carriers. MCE covalently immobilized on activated chitosan polymer with the bifunctional agent glutaraldehyde (Ch-MCE) exhibited highest immobilization yield (74.6 %). Comparing to the native MCE, Ch-MCE exhibited higher optimum pH, higher optimum reaction temperature, lower activation energy, higher half-life time, lower deactivation rate constant and higher energy for denaturation. After immobilization, maximum reaction rate, Michaelis-Menten constant, specificity constant, turnover number, and catalytic efficiency of the enzyme were significantly changed. Calculated thermodynamic parameters for denaturation (enthalpy, entropy and Gibbs free energy) confirmed that the catalytic properties of MCE were significantly improved after immobilization. Reusability tests showed that after 7 catalytic cycles, the Ch-MCE retained about 71 % of its activity confirming its suitability for industrial applications.

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Catalytic, thermodynamic and structural properties of an immobilized and highly thermostable alkaline protease from a haloalkaliphilic actinobacteria, Nocardiopsis alba TATA-5

Cited by 50 publications

References 37 publications

Biochemical properties of extracellular protease from Staphylococcus carnosus RT6 isolated from Harbin dry sausages, and its hydrolysis of meat proteins

Biochemical properties of extracellular protease from Staphylococcus carnosus RT6 isolated from Harbin dry sausages, and its hydrolysis of meat proteins

Cloning, Expression, and Structural Elucidation of a Biotechnologically Potential Alkaline Serine Protease From a Newly Isolated Haloalkaliphilic Bacillus lehensis JO-26

Kinetic, Catalytic and Thermodynamic Properties of Immobilized Milk Clotting Enzyme on Activated Chitosan Polymer and its Application in Cheese Making

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