Biochemical and molecular characterization of a thermo- and detergent-stable alkaline serine keratinolytic protease from Bacillus circulans strain DZ100 for detergent formulations and feather-biodegradation process

被引：72

作者：

Benkiar, Amina ^{[1
,2
]}

Nadia, Zarai Jaouadi ^{[3
]}

Badis, Abdelmalek ^{[1
,2
]}

Rebzani, Feriel ^{[1
]}

Soraya, Boulkour Touioui ^{[1
]}

Rekik, Hatem ^{[3
]}

Naili, Belgacem ^{[3
]}

Ferradji, Fatma Zohra ^{[1
,2
]}

Bejar, Samir ^{[3
]}

Jaouadi, Bassem ^{[3
]}

机构：

[1] Univ Saad Dahlab Blida, LNPCB, Blida 09000, Algeria

[2] Natl Ctr Res & Dev Fisheries & Aquaculture CNRDPA, Bousmail, W Tipaza, Algeria

[3] Univ Sfax, Ctr Biotechnol Sfax, LMB, Sfax 3018, Tunisia

来源：

INTERNATIONAL BIODETERIORATION & BIODEGRADATION | 2013年 / 83卷

关键词：

Bacillus circulans; Protease; Thermostability; Detergent; Feather-biodegradation; PUMILUS CBS; PURIFICATION; PROTEINASE; KERATINASE; IDENTIFICATION; SAPB;

D O I：

10.1016/j.ibiod.2013.05.014

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

An extracellular keratinolytic protease (SAPDZ) was produced and purified from a newly isolated Bacillus circulans strain DZ100. Matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF/MS) analysis revealed that the purified enzyme is a monomer with a molecular mass of 32019.10 Da. The sequence of the 25 N-terminal residues of SAPDZ showed high homology with those of Bacillus proteases. Optimal activity was achieved at pH 12.5 and 85 degrees C. This enzyme was completely inhibited by phenylmethanesulfonyl fluoride (PMSF) and diiodopropyl fluorophosphates (DFP), which suggests that it belongs to the serine protease family. Compared to the other tested proteases, SAPDZ exhibited broader substrate specificity, higher levels of catalytic efficiency, and greater keratinolytic activity, which made it able to accomplish the entire feather-biodegradation process on its own. The sapDZ gene encoding SAPDZ was cloned, sequenced, and expressed in Escherichia coli. The biochemical properties exhibited by the extracellular purified recombinant enzyme (rSAPDZ) were similar to those of the native one. Above all, SAPDZ exhibited marked stability to detergents, making it a potential candidate for future applications in detergent formulations and an efficient eco-friendly enzyme for the biodegradation of feather keratin. (C) 2013 Elsevier Ltd. All rights reserved.

引用

页码：129 / 138

页数：10

共 23 条

[1] Purification and characterization of an oxidation-stable, thiol-dependent serine alkaline protease from Bacillus mojavensis [J].

Beg, QK ;

Gupta, R .

ENZYME AND MICROBIAL TECHNOLOGY, 2003, 32 (02) :294-304

[2]

BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3

[3] TISSUE SULFHYDRYL GROUPS [J].

ELLMAN, GL .

ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1959, 82 (01) :70-77

[4] GROWTH, KERATINOLYTIC PROTEINASE ACTIVITY AND THERMOTOLERANCE OF DERMATOPHYTES ASSOCIATED WITH ALOPECIA IN UYO, NIGERIA [J].

Essien, J. P. ;

Umoh, A. A. ;

Akpan, E. J. ;

Eduok, S. I. ;

Umoiyoho, A. .

ACTA MICROBIOLOGICA ET IMMUNOLOGICA HUNGARICA, 2009, 56 (01) :61-69

[5] Bacterial alkaline proteases: molecular approaches and industrial applications [J].

Gupta, R ;

Beg, QK ;

Lorenz, P .

APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2002, 59 (01) :15-32

[6] Biochemical and molecular characterization of a detergent-stable serine alkaline protease from Bacillus pumilus CBS with high catalytic efficiency [J].

Jaouadi, Bassem ;

Ellouz-Chaabouni, Semia ;

Rhimi, Moez ;

Bejar, Samir .

BIOCHIMIE, 2008, 90 (09) :1291-1305

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

Jaouadi, Bassem ;

Abdelmalek, Badis ;

Fodil, Djamila ;

Ferradji, Fatma Zohra ;

Rekik, Hatem ;

Zarai, Nedia ;

Bejar, Samir .

BIORESOURCE TECHNOLOGY, 2010, 101 (21) :8361-8369

[8] Excellent Laundry Detergent Compatibility and High Dehairing Ability of the Bacillus pumilus CBS Alkaline Proteinase (SAPB) [J].

Jaouadi, Bassem ;

Ellouz-Chaabouni, Semia ;

Ali, Mamdouh Ben ;

Messaoud, Ezzedine Ben ;

Naili, Belgacem ;

Dhouib, Abdelhafidh ;

Bejar, Samir .

BIOTECHNOLOGY AND BIOPROCESS ENGINEERING, 2009, 14 (04) :503-512

[9] The overexpression of the SAPB of Bacillus pumilus CBS and mutated sapB-L31I/T33S/N99Y alkaline proteases in Bacillus subtilis DB430: New attractive properties for the mutant enzyme [J].

Jaouadi, Nadia Zarai ;

Jaouadi, Bassem ;

Aghajari, Nushin ;

Bejar, Samir .

BIORESOURCE TECHNOLOGY, 2012, 105 :142-151

[10] Characterization of an alkaline active thiol forming extracellular serine keratinase by the newly isolated Bacillus pumilus [J].

Kumar, A. Ganesh ;

Swarnalatha, S. ;

Gayathri, S. ;

Nagesh, N. ;

Sekaran, G. .

JOURNAL OF APPLIED MICROBIOLOGY, 2008, 104 (02) :411-419

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