Purification and characterization of a thermostable keratinolytic serine alkaline proteinase from Streptomyces sp. strain AB1 with high stability in organic solvents

Jaouadi, Bassem; Badis, Abdelmalek; Fodil, Djamila; Ferradji, Fatma Zohra; Rekik, Hatem; Zaraî, Nedia; Béjar, Samír

doi:10.1016/j.biortech.2010.05.066

Cited by 118 publications

(81 citation statements)

References 31 publications

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“…Potential and prospects for SAPB and KERAB in the leather processing industry 6.1 Keratin-degradation profile of Bacillus pumilus CBS and Streptomyces sp. AB1 Keratinacious substrates, such as keratin and keratin azure, were previously reported to be significantly hydrolyzed by SAPB and KERAB (Jaouadi et al, 2010a). It was also demonstrated that the B. pumilus strain CBS was able to grow in an optimized medium containing 10 g/l of feather-meal, chicken feather (Fig.…”

Section: Storage Stability Of the Spray-dried And Lyophilized Sapbmentioning

confidence: 80%

“…For determination of their molecular weight, enzyme preparations were treated with 1 mM PMSF prior to electrophoresis to inhibit possible autolysis during electrophoresis. Electrophoresis under denaturing conditions (SDS-PAGE) also revealed single bands with molecular masses estimated as 34 kDa for SAPB and 30 kDa for KERAB (Jaouadi et al, 2010a). The exact molecular masses obtained for the purified SAPB and KERAB were confirmed by MALDI-TOF mass spectrometry as being 34598.19 and 29850.17 Da, respectively.…”

Section: Sapb and Kerab Purification And Characterizationmentioning

confidence: 82%

“…In medium containing trace salts with feather as carbon and nitrogen source (g/l): NaCl, 0.5; KH 2 PO 4 , 0.5; K 2 HPO 4 , 0.5; KCl, 0.1; MgSO 4 ·7H 2 O, 1; and chicken feather meal, 10; at pH 9, KERAB production was observed to undergo a significant improvement, reaching a maximum of 9,500 U/ml under the optimal conditions used (pH 11.5 and 75°C) after 96 h of incubation at 30°C and 200 rpm (Jaouadi et al, 2010a). Under these particular conditions, the production of the SAPB and KERAB enzymes started after a 6-and 10-h lag phase,…”

Section: Strain Ab1 (Gu434672)mentioning

confidence: 96%

“…The purity of the enzyme was estimated to be about 86-fold greater than that of the crude extract. The purified enzyme preparation contained about 24% of the total activity of the crude enzyme and had a specific activity of 67,000 U/mg (Jaouadi et al, 2010a). These preparations were homogeneous enzymes with high purity as they exhibited single protein bands on native PAGE and unique elution symmetrical peaks on gel filtration chromatography.…”

Section: Sapb and Kerab Purification And Characterizationmentioning

confidence: 99%

“…A molecular approach was used to establish further support for the identification of the CBS and AB1 isolates. Two 16S rRNA gene fragments, namely 1,497 bp (Jaouadi et al, 2009) and 1541 bp (Jaouadi et al, 2010a), were amplified from the genomic DNA of the CBS and AB1 isolates, respectively, and then cloned and sequenced on both strands. The 16S rRNA gene sequences obtained were subjected to GenBank BLAST search analyses, which yielded strong homologies of up to 98 and 99% with those of several cultivated strains of Bacillus and Streptomyces, respectively.…”

Section: Identification and Molecular Phylogeny Of The Microorganismsmentioning

confidence: 99%

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The Bioengineering and Industrial Applications of Bacterial Alkaline Proteases: the Case of SAPB and KERAB

Jaouadi¹,

Badis²,

Jaouadi³

et al. 2011

Progress in Molecular and Environmental Bioengineering - From Analysis and Modeling to Technology Applications

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Section: Storage Stability Of the Spray-dried And Lyophilized Sapbmentioning

confidence: 80%

Section: Sapb and Kerab Purification And Characterizationmentioning

confidence: 82%

Section: Strain Ab1 (Gu434672)mentioning

confidence: 96%

Section: Sapb and Kerab Purification And Characterizationmentioning

confidence: 99%

Section: Identification and Molecular Phylogeny Of The Microorganismsmentioning

confidence: 99%

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The Bioengineering and Industrial Applications of Bacterial Alkaline Proteases: the Case of SAPB and KERAB

Jaouadi¹,

Badis²,

Jaouadi³

et al. 2011

Progress in Molecular and Environmental Bioengineering - From Analysis and Modeling to Technology Applications

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Cloning and characterization of a thermostable detergent‐compatible recombinant keratinase from Bacillus pumilus KS12

Rajput

Sharma

Gupta

2011

Biotech and App Biochem

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Abstract.Functional expression of a keratinase from a potential feather-degrading bacterium, Bacillus pumilus KS12, was achieved in Escherichia coli using pEZZ18 vector. The enzyme was constitutively secreted at 37• C and 300 rpm after 18 H of incubation and was purified to homogeneity using diethylaminoethyl-Sepharose column. It was completely stable within the pH range of 7.0-10.0 with optima at pH 9.0, and temperature 30• C-90• C with optima at 70 • C. It had high thermostability with a t 1/2 of more than 4 H at 70• C, more than 2 H at 80• C, and 30 Min at 90• C. The enzyme was identified as a serine hydrolase as it was completely inhibited by 10 mM phenylmethylsulfonyl fluoride. It retained more than 90% relative activity in the presence of chelating agents such as ethylenediaminetetraacetic acid and 1,10-o-phenanthroline. It was also a thiol-activated enzyme with 3.32-and fourfold activation in the presence of 10 mM dithiothreitol and β-mercaptoethanol. The keratinase exhibited high detergent compatibility and oxidation stability with an eight-and fivefold enhancement in the presence of triton X-100 and saponin, respectively. It hydrolyzed a large array of protein substrates with a keratinolytic-caseinolytic ratio of more than 0.5. Amidolytic activity revealed its cleavage on phenylalanine → leucine → alanine-p-nitroanilides. In addition, electrospray ionization-mass spectrometry analysis of hydrolyzed insulin B-chain revealed cleavage between Leu 6 -Cys 7 , Cys 7 -Gly 8 , Tyr 16 -Leu

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Purification, characterization and potential applications of a novel halotolerant metalloprotease from marine bacterium Vibrio sp. LA‐05

Zhang

Lang

et al. 2018

J of Chemical Tech & Biotech

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BACKGROUND: Many reported proteases showed low activity and stability under harsh operating conditions, e.g. high pH, salinity and surfactants, which hinders expansion of their applications in industry. Here, a novel halotolerant metalloprotease from marine bacterium Vibrio sp. LA-05 endowed with superior properties was purified and characterized, and its industrial application also evaluated. RESULTS:The protease was active and stable at 25-40 ∘ C and pH 6.0-10.0. Moreover, it was remarkably stable in high salt solution, tolerant to denaturing agents, organic solvents, surfactants and bleaching agents, and quite compatible and stable with many commercial detergents. Its residual activities were above 22% even after co-incubation with 20% NaCl for 480 h. Except Ariel and Tide, the residual activities were all above 55% after co-incubation with five other liquid detergents for 240 h. The corresponding theoretical analyses show that it is a metalloprotease containing 1 mol of zinc ion per mole of protease, the mature active protease predominantly in -helix form consists of 321 amino acids, and the central zinc ion along with 11 other conserved and crucial amino acid residues forms the catalytic domain. CONCLUSION: These excellent properties indicated that the protease has considerable potential as a bio-additive in the application of detergent formulations.

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Purification and characterization of a thermostable keratinolytic serine alkaline proteinase from Streptomyces sp. strain AB1 with high stability in organic solvents

Cited by 118 publications

References 31 publications

The Bioengineering and Industrial Applications of Bacterial Alkaline Proteases: the Case of SAPB and KERAB

The Bioengineering and Industrial Applications of Bacterial Alkaline Proteases: the Case of SAPB and KERAB

Cloning and characterization of a thermostable detergent‐compatible recombinant keratinase from Bacillus pumilus KS12

Purification, characterization and potential applications of a novel halotolerant metalloprotease from marine bacterium Vibrio sp. LA‐05

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