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( Silveira Silvana T. ),( Sabrine Gemelli ),( Jeferson Segalin ),( Adriano Brandelli ) 한국미생물 · 생명공학회 2012 Journal of microbiology and biotechnology Vol.22 No.6
Keratinases are exciting keratin-degrading enzymes; however, there have been relatively few studies on their immobilization. A keratinolytic protease from Chryseobacterium sp. kr6 was purified and its partial sequence determined using mass spectrometry. No significant homology to other microbial peptides in the NCBI database was observed. Certain parameters for immobilization of the purified keratinase on chitosan beads were investigated. The production of the chitosan beads was optimized using factorial design and surface response techniques. The optimum chitosan bead production for protease immobilization was a 20 g/l chitosan solution in acetic acid [1.5% (v/v)], glutaraldehyde ranging from 34 g to 56 g/l, and an activation time between 6 and 10 h. Under these conditions, above 80% of the enzyme was immobilized on the support. The behavior of the keratinase loading on the chitosan beads surface was well described using the Langmuir model. The maximum capacity of the support (qm) and dissociation constant (Kd) were estimated as 58.8 U/g and 0.245 U/ml, respectively. The thermal stability of the immobilized enzyme was also improved around 2-fold, when compared with that of the free enzyme, after 30 min at 65oC. In addition, the activity of the immobilized enzyme remained at 63.4% after it was reused five times. Thus, the immobilized enzyme exhibited an improved thermal stability and remained active after several uses.
Pectin from Passion Fruit Fiber and Its Modification by Pectinmethylesterase
Contreras-Esquivel, Juan Carlos,Aguilar, Cristobal N.,Montanez, Julio C.,Brandelli, Adriano,Espinoza-Perez, Judith D.,Renard, Catherine M.G.C. The Korean Society of Food Science and Nutrition 2010 Preventive Nutrition and Food Science Vol.15 No.1
Passion fruit fiber pectin gels represent a new alternative pectin source with potential for food and non-food applications on a commercial scale. Pectic polysaccharides were extracted from passion fruit (Passiflora edulis) fiber using citric acid as a clean catalyst and autoclaved for 20 to 60 min at $121^{\circ}C$. The best condition of pectin yield with the highest molecular weight was obtained with 1.0% of citric acid (250 mg/g dry passion fruit fiber pectin) for 20 min of autoclaving. Spectroscopic analyses by Fourier transform infrared, enzymatic degradation reactions, and ion-exchange chromatography assays showed that passion fruit pectin extracted for 20 min was homogeneous high methoxylated pectin (70%). Gel permeation analysis confirmed that the pectin extract obtained by autoclaving by 20 min showed higher molecular weights than those autoclaved for 40 and 60 min. Passion fruit pectin extracted for 20 min was enzymatically modified with fungal pectinmethylesterase to create restructured gels. Short autoclave treatment (20 min) with citric acid as extractant resulted in a significant increase of gel strength, improving pectin extraction in terms of functionality. The treatment of solubilized material (pectic polysaccharides) in the presence of insoluble material (cellulose and hemicellulose) with pectinmethylesterase and calcium led to the creation of a stiffer passion fruit fiber pectin gel, while syneresis was not observed.
Pectin from Passion Fruit Fiber and Its Modification by Pectinmethylesterase
Juan Carlos Contreras-Esquivel,Cristobal N. Aguilar,Julio C. Montanez,Adriano Brandelli,Judith D. Espinoza-Perez,Catherine M.G.C. Renard 한국식품영양과학회 2010 Preventive Nutrition and Food Science Vol.15 No.1
Passion fruit fiber pectin gels represent a new alternative pectin source with potential for food and non-food applications on a commercial scale. Pectic polysaccharides were extracted from passion fruit (Passiflora edulis) fiber using citric acid as a clean catalyst and autoclaved for 20 to 60 min at 121℃. The best condition of pectin yield with the highest molecular weight was obtained with 1.0% of citric acid (250 ㎎/g dry passion fruit fiber pectin) for 20 min of autoclaving. Spectroscopic analyses by Fourier transform infrared, enzymatic degradation reactions, and ion-exchange chromatography assays showed that passion fruit pectin extracted for 20 min was homogeneous high methoxylated pectin (70%). Gel permeation analysis confirmed that the pectin extract obtained by autoclaving by 20 min showed higher molecular weights than those autoclaved for 40 and 60 min. Passion fruit pectin extracted for 20 min was enzymatically modified with fungal pectinmethylesterase to create restructured gels. Short autoclave treatment (20 min) with citric acid as extractant resulted in a significant increase of gel strength, improving pectin extraction in terms of functionality. The treatment of solubilized material (pectic polysaccharides) in the presence of insoluble material (cellulose and hemicellulose) with pectinmethylesterase and calcium led to the creation of a stiffer passion fruit fiber pectin gel, while syneresis was not observed.
Isolation and Characterization of Antifungal Peptides Produced by Bacillus amyloliquefaciens LBM5006
Lisianne Brittes Benitez,Renata Voltolini Velho,Marcia Pagno Lisboa,Luis Fernando da Costa Medina,Adriano Brandelli 한국미생물학회 2010 The journal of microbiology Vol.48 No.6
Bacillus amyloliquefaciens LBM 5006 produces antagonistic activity against pathogenic bacteria and phytopathogenic fungi, including Aspergillus spp., Fusarium spp., and Bipolaris sorokiniana. PCR analysis revealed the presence of ituD, but not sfp genes, coding for iturin and surfactin, respectively. The antimicrobial substance produced by this strain was isolated by ammonium sulfate precipitation, gel filtration chromatography and 1-butanol extraction. The ultraviolet spectrum was typical of a polypeptide and the infrared spectrum indicates the presence of peptide bonds and acyl group(s). The antimicrobial substance was resistant to proteolytic enzymes and heat treatment, and was reactive with ninhydrin. Mass spectroscopy analysis indicated that B. amyloliquefaciens LBM 5006 produces two antimicrobial peptides,with main peaks at m/z 1,058 Da and 1,464 Da, corresponding to iturin-like and fengycin-like peptides,respectively. B. amyloliquefaciens LBM 5006 showed significant activity against phytopatogenic fungi, showing potential for use as a biocontrol agent or production of antifungal preparations.