Intracellular alkaline proteases produced by thermoacidophiles: detection of protease heterogeneity by gelatin zymography and polymerase chain reaction (PCR)

Kocabiyik, Semra; Erdem, Bilge

doi:10.1016/s0960-8524(02)00019-6

Cited by 21 publications

(11 citation statements)

References 17 publications

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“…The protease activity was determined by using method suggested by Kocabiyik and Erdem (2002) with minor modification. One unit of protease activity on azocasein was defined as the absorbance change of 0.1 per min at 440 nm under the standard assay condition (Blumentals et al, 1990).…”

Section: Assay Of Protease Activitymentioning

confidence: 99%

“…Cereghino and Cregg (2000) reported that P. pastoris expression system was able to produce heterologous proteins range from 10-100 fold higher than their native hosts. The positive transformants from PCR analysis were cultivated and tested for protease expression by using method described by previous study (Fu, 2001;Kocabiyik and Erdem, 2002). The enzyme was preincubated at 70°C prior to assay in order to activate the F1 protease and eliminate other native proteins from the host.…”

Section: Transformation Into P Pastoris Strain Gs115 and Smd1168hmentioning

confidence: 99%

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Secretory expression of thermostable alkaline protease from Bacillus stearothermophilus FI by using native signal peptide and α-factor secretion signal in Pichia pastoris

et al. 2013

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The thermostable alkaline protease from Bacillus stearothermophilus F1 has high potential for industrial applications, and attempt to produce the enzyme in yeast for higher yield was undertaken. Secretory expression of F1 protease through yeast system could improve enzyme's capability, thus simplifying the purification steps. Mature and full genes of F1 protease were cloned into Pichia pastoris expression vectors (pGAPZαB and pPICZαB) and transformed into P. pastoris strains (GS115 and SMD1168H) via electroporation method. Recombinant F1 protease under regulation constitutive GAP promoter revealed that the highest expression was achieved after 72 h cultivation. While inducible AOX promoter showed that 0.5% (v/v) methanol was the best to induce expression. It was proven that constitutive expression strategy was better than inducible system. The α-secretion signal from the plasmid demonstrated higher secretory expression level of F1 protease as compared to native Open Reading Frame (ORF) in GS115 strain (GE6GS). Production medium YPTD was found to be the best for F1 protease expression with the highest yield of 4.13 U/mL. The protein was expressed as His-tagged fusion protein with a size about 34 kDa.

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Section: Assay Of Protease Activitymentioning

confidence: 99%

Section: Transformation Into P Pastoris Strain Gs115 and Smd1168hmentioning

confidence: 99%

Secretory expression of thermostable alkaline protease from Bacillus stearothermophilus FI by using native signal peptide and α-factor secretion signal in Pichia pastoris

et al. 2013

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“…Alkaliphiles enzymes have ability to resistant high concentration of detergents and showing their optimum activity at high pH such as lipases, amylases, and proteases. Based on PCR homology combinations, the alkaline proteases have been isolated from hot environments by collecting thermoacidophilic bacteria and archaea [46]. DOI: 10.9790/3008-1201021017 www.iosrjournals.org 13 | Page…”

Section: Alkalismentioning

confidence: 99%

Extremophile Current Challenges and New Gate of Knowledge by Nanoparticles Pathways.

Moayad¹,

Zha²,

Yan³

2017

IOSRJPBS

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“…19 We used an initial enzyme solution with concentration 20 U/mL, and the total volume was 100 mL. The fabric sample was added and left standing for 10 min.…”

Section: Removal and Deactivation Of The Enzymementioning

confidence: 99%

A study on using of protease for removal of animal glue adhesive in textile conservation

Ahmed

Kolisis

2011

J of Applied Polymer Sci

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Animal glue has been used to fix historical textiles on paper, wood panels, or other rigid support materials. It is often present in shrunk, cracked, rigid, and brittle form because of the aged condition artifacts and may not provide enough adhesion for effective support causing damage to historical textiles. The biotechnological application of enzymes seems to be a very promising approach in the restoration of historical objects. In this experimental work, undertaken with modern linen and silk fabrics, interesting results have been obtained for the removal of animal glue by using the protease enzyme from Aspergillus oryzae. An extensive study was done in the enzymatic activity and efficiency for the removal of the animal glue from the textiles, as well as the effects of this treatment on mechanical and optical parameters of the textile fibers. The effect of protease on fibers is measured by Fourier transform infrared spectral analysis, scanning electron microscope, the CIE-Lab values, ASTM method D5035, and XRD. The results showed that using protease in adhesive removal presented good results with a safe and a short treatment time when compared with the conventional methods. No significant changes on the linen and silk fabrics are observed.

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Intracellular alkaline proteases produced by thermoacidophiles: detection of protease heterogeneity by gelatin zymography and polymerase chain reaction (PCR)

Cited by 21 publications

References 17 publications

Secretory expression of thermostable alkaline protease from <i>Bacillus stearothermophilus</i> FI by using native signal peptide and <b>α</b>-factor secretion signal in <i>Pichia pastoris</i>

Secretory expression of thermostable alkaline protease from <i>Bacillus stearothermophilus</i> FI by using native signal peptide and <b>α</b>-factor secretion signal in <i>Pichia pastoris</i>

Extremophile Current Challenges and New Gate of Knowledge by Nanoparticles Pathways.

A study on using of protease for removal of animal glue adhesive in textile conservation

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