Ostafe, V.

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  • Ostafe, V. (2)
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Author's Bibliography

Biochemical characterization of chitinase A from Bacillus licheniformis DSM8785 expressed in Pichia pastoris KM71H

Menghiu, G.; Ostafe, V.; Prodanović, Radivoje; Fischer, Rainer; Ostafe, Raluca

(2019)

TY  - JOUR
AU  - Menghiu, G.
AU  - Ostafe, V.
AU  - Prodanović, Radivoje
AU  - Fischer, Rainer
AU  - Ostafe, Raluca
PY  - 2019
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/2798
AB  - Chitin is an abundant biopolymer found mainly in the exoskeleton of crustaceans and insects. The degradation of chitin using chitinases is one way to address the accumulation of chitin waste streams in the environment, and research has therefore focused on the identification, improvement and expression of suitable enzymes. Here we describe the production, purification and characterization of Bacillus licheniformis chitinase A in the Pichia pastoris expression system. Optimal enzyme activity occurred at pH 4.0–5.0 and within the temperature range 50–60 °C. With colloidal chitin as the substrate, the Km (2.307 mM) and Vmax (0.024 mM min−1) of the enzyme were determined using a 3,5-dinitrosalicylic acid assay. The degradation products of colloidal chitin and hexa-N-acetylchitohexaose were compared by thin-layer chromatography. The activity of the glycosylated enzyme produced in P. pastoris was compared with the in vitro deglycosylated and aglycosylated version produced in Escherichia coli. We showed that the glycosylated chitinase was more active than the deglycosylated and aglycosylated variants. © 2018 Elsevier Inc.
T2  - Protein Expression and Purification
T1  - Biochemical characterization of chitinase A from Bacillus licheniformis DSM8785 expressed in Pichia pastoris KM71H
VL  - 154
SP  - 25
EP  - 32
DO  - 10.1016/j.pep.2018.09.007
ER  - 
@article{
author = "Menghiu, G. and Ostafe, V. and Prodanović, Radivoje and Fischer, Rainer and Ostafe, Raluca",
year = "2019",
url = "http://cherry.chem.bg.ac.rs/handle/123456789/2798",
abstract = "Chitin is an abundant biopolymer found mainly in the exoskeleton of crustaceans and insects. The degradation of chitin using chitinases is one way to address the accumulation of chitin waste streams in the environment, and research has therefore focused on the identification, improvement and expression of suitable enzymes. Here we describe the production, purification and characterization of Bacillus licheniformis chitinase A in the Pichia pastoris expression system. Optimal enzyme activity occurred at pH 4.0–5.0 and within the temperature range 50–60 °C. With colloidal chitin as the substrate, the Km (2.307 mM) and Vmax (0.024 mM min−1) of the enzyme were determined using a 3,5-dinitrosalicylic acid assay. The degradation products of colloidal chitin and hexa-N-acetylchitohexaose were compared by thin-layer chromatography. The activity of the glycosylated enzyme produced in P. pastoris was compared with the in vitro deglycosylated and aglycosylated version produced in Escherichia coli. We showed that the glycosylated chitinase was more active than the deglycosylated and aglycosylated variants. © 2018 Elsevier Inc.",
journal = "Protein Expression and Purification",
title = "Biochemical characterization of chitinase A from Bacillus licheniformis DSM8785 expressed in Pichia pastoris KM71H",
volume = "154",
pages = "25-32",
doi = "10.1016/j.pep.2018.09.007"
}
Menghiu, G., Ostafe, V., Prodanović, R., Fischer, R.,& Ostafe, R. (2019). Biochemical characterization of chitinase A from Bacillus licheniformis DSM8785 expressed in Pichia pastoris KM71H.
Protein Expression and Purification, 154, 25-32.
https://doi.org/10.1016/j.pep.2018.09.007
Menghiu G, Ostafe V, Prodanović R, Fischer R, Ostafe R. Biochemical characterization of chitinase A from Bacillus licheniformis DSM8785 expressed in Pichia pastoris KM71H. Protein Expression and Purification. 2019;154:25-32
Menghiu G., Ostafe V., Prodanović Radivoje, Fischer Rainer, Ostafe Raluca, "Biochemical characterization of chitinase A from Bacillus licheniformis DSM8785 expressed in Pichia pastoris KM71H" Protein Expression and Purification, 154 (2019):25-32,
https://doi.org/10.1016/j.pep.2018.09.007 .
11
8
9

Biochemical characterization of chitinase A from Bacillus licheniformis DSM8785 expressed in Pichia pastoris KM71H

Menghiu, G.; Ostafe, V.; Prodanović, Radivoje; Fischer, Rainer; Ostafe, Raluca

(Elsevier, 2019)

TY  - JOUR
AU  - Menghiu, G.
AU  - Ostafe, V.
AU  - Prodanović, Radivoje
AU  - Fischer, Rainer
AU  - Ostafe, Raluca
PY  - 2019
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/348
AB  - Chitin is an abundant biopolymer found mainly in the exoskeleton of crustaceans and insects. The degradation of chitin using chitinases is one way to address the accumulation of chitin waste streams in the environment, and research has therefore focused on the identification, improvement and expression of suitable enzymes. Here we describe the production, purification and characterization of Bacillus licheniformis chitinase A in the Pichia pastoris expression system. Optimal enzyme activity occurred at pH 4.0–5.0 and within the temperature range 50–60 °C. With colloidal chitin as the substrate, the Km (2.307 mM) and Vmax (0.024 mM min−1) of the enzyme were determined using a 3,5-dinitrosalicylic acid assay. The degradation products of colloidal chitin and hexa-N-acetylchitohexaose were compared by thin-layer chromatography. The activity of the glycosylated enzyme produced in P. pastoris was compared with the in vitro deglycosylated and aglycosylated version produced in Escherichia coli. We showed that the glycosylated chitinase was more active than the deglycosylated and aglycosylated variants. © 2018 Elsevier Inc.
PB  - Elsevier
T2  - Protein Expression and Purification
T1  - Biochemical characterization of chitinase A from Bacillus licheniformis DSM8785 expressed in Pichia pastoris KM71H
VL  - 154
SP  - 25
EP  - 32
DO  - 10.1016/j.pep.2018.09.007
ER  - 
@article{
author = "Menghiu, G. and Ostafe, V. and Prodanović, Radivoje and Fischer, Rainer and Ostafe, Raluca",
year = "2019",
url = "http://cherry.chem.bg.ac.rs/handle/123456789/348",
abstract = "Chitin is an abundant biopolymer found mainly in the exoskeleton of crustaceans and insects. The degradation of chitin using chitinases is one way to address the accumulation of chitin waste streams in the environment, and research has therefore focused on the identification, improvement and expression of suitable enzymes. Here we describe the production, purification and characterization of Bacillus licheniformis chitinase A in the Pichia pastoris expression system. Optimal enzyme activity occurred at pH 4.0–5.0 and within the temperature range 50–60 °C. With colloidal chitin as the substrate, the Km (2.307 mM) and Vmax (0.024 mM min−1) of the enzyme were determined using a 3,5-dinitrosalicylic acid assay. The degradation products of colloidal chitin and hexa-N-acetylchitohexaose were compared by thin-layer chromatography. The activity of the glycosylated enzyme produced in P. pastoris was compared with the in vitro deglycosylated and aglycosylated version produced in Escherichia coli. We showed that the glycosylated chitinase was more active than the deglycosylated and aglycosylated variants. © 2018 Elsevier Inc.",
publisher = "Elsevier",
journal = "Protein Expression and Purification",
title = "Biochemical characterization of chitinase A from Bacillus licheniformis DSM8785 expressed in Pichia pastoris KM71H",
volume = "154",
pages = "25-32",
doi = "10.1016/j.pep.2018.09.007"
}
Menghiu, G., Ostafe, V., Prodanović, R., Fischer, R.,& Ostafe, R. (2019). Biochemical characterization of chitinase A from Bacillus licheniformis DSM8785 expressed in Pichia pastoris KM71H.
Protein Expression and Purification
Elsevier., 154, 25-32.
https://doi.org/10.1016/j.pep.2018.09.007
Menghiu G, Ostafe V, Prodanović R, Fischer R, Ostafe R. Biochemical characterization of chitinase A from Bacillus licheniformis DSM8785 expressed in Pichia pastoris KM71H. Protein Expression and Purification. 2019;154:25-32
Menghiu G., Ostafe V., Prodanović Radivoje, Fischer Rainer, Ostafe Raluca, "Biochemical characterization of chitinase A from Bacillus licheniformis DSM8785 expressed in Pichia pastoris KM71H" Protein Expression and Purification, 154 (2019):25-32,
https://doi.org/10.1016/j.pep.2018.09.007 .
11
8
9