Production and properties of an alkaline protease by Aureobasidium pullulans

Donaghy, John; McKay, A.M.

doi:10.1111/j.1365-2672.1993.tb05200.x

Cited by 44 publications

(17 citation statements)

References 25 publications

(22 reference statements)

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“…The enhancement of protease activity in presence of s-block metals (Ca 2+ , Mg 2+ ) and reduction in the presence of d-block metals (Zn 2+ , Cu 2+ , Hg 2+ , Cr 3+ ) has been reported by many researchers (Oberoi et al, 2001;Patel et al, 2006;Arulmani et al, 2007;Jaouadi et al, 2008;Shah et al, 2010). This result suggests that the concerned metal ions protected the enzyme against thermal denaturation and played vital role in maintaining active confirmation of the enzyme at high temperatures (Donaghy and Mckay, 1993). Generally, s-block metals bind poorly to ligands through ionic bond.…”

supporting

confidence: 48%

Purification and partial characterization of a detergent and oxidizing agent stable alkaline protease from a newly isolated Bacillus subtilis VSG-4 of tropical soil

et al. 2011

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An extracellular detergent tolerant protease producing strain VSG-4 was isolated from tropical soil sample and identified as Bacillus subtilis based on morphological, biochemical characteristics as well as 16S-rRNA gene sequencing. The VSG-4 protease was purified to homogeneity using ammonium sulphate precipitation, dialysis and sephadex G-200 gel permeation chromatography with a 17.4 purification fold. The purified enzyme was active and stable over a broad range of pH (8.0-11.0, optimum at 9.0) and temperature (40°C to 60°C, optimum at 50°C). The thermostability of the enzyme was significantly increased by the addition CaCl(2). This enzyme was strongly inhibited by PMSF and DFP, suggesting that it belongs to the serine protease superfamily. The purified VSG-4 alkaline protease showed remarkable stability in anionic (5 mM SDS) and ionic (1% Trion X-100 and 1% Tween 80) detergents. It retained 97±2% and 83.6±1.1% of its initial activity after 1 h preincubation in the presence of 1 % H(2)O(2) and 1 % sodium perborate, respectively. Furthermore, the purified enzyme showed excellent stability and compatibility with some commercial laundry detergents besides its stain removal capacity. Considering these promising properties, VSG-4 protease may find tremendous application in laundry detergent formulations.

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confidence: 48%

Purification and partial characterization of a detergent and oxidizing agent stable alkaline protease from a newly isolated Bacillus subtilis VSG-4 of tropical soil

et al. 2011

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“…Besides, these cations were also reported to enhance the thermal stability of other alkaline protease from Bacillus sp. by protecting the enzyme against thermal denaturation and maintaining the activity conformation of the enzyme at high temperature [31,32]. The metal chelators (EDTA and EGTA) could strongly inhibit the enzyme.…”

Section: Discussionmentioning

confidence: 99%

Purification and Characterization of a Novel Collagenase from Bacillus pumilus Col-J

et al. 2009

Appl Biochem Biotechnol

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The collagenase, produced extracellular by Bacillus pumilus Col-J, was purified by ammonium sulfate precipitation followed by two gel filtrations, involving Sephadex G-100 column and Sepharose Fast Flow column. Purified collagenase has a 31.53-fold increase in specific activity of 87.33 U/mg and 7.00% recovery. The collagenase has a relative molecular weight of 58.64 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The optimal temperature for the enzyme reaction was 45 degrees C. More than 50% of the original activity still remained after 5 min of incubation at 70 degrees C or 10 min at 60 degrees C. The maximal enzyme activity of collagenase was obtained at pH 7.5, and it was stable over a pH range of 6.5-8.0. The collagenase activity was strongly inhibited by Mn(2+), Pb(2+), ethylenediamine tetraacetic acid, ethylene glycol tetraacetic acid, and beta-mercaptoethanol. However, Ca(2+) and Mg(2+) greatly increased its activity. The collagenase from B. pumilus Col-J showed highly specific activity towards the native collagen from calf skin. The K(m) and V(max) of the enzyme for collagen were 0.79 mg/mL and 129.5 U, respectively.

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“…In general, cations protect the enzyme against thermal denaturation and play a vital role in maintaining the active conformation of the enzyme at high temperatures [10,49]. The AH-6 protease was exceptionally stable in the presence of heavy metal, HgCl 2 where the enzyme retained 100 and 50% of the original activity at 10 and 100 mM HgCl 2 , respectively.…”

Section: Effect Of Cations On Protease Activitymentioning

confidence: 99%

Purification and stability characteristics of an alkaline serine protease from a newly isolated Haloalkaliphilic bacterium sp. AH-6

Dodia¹,

Rawal²,

Bhimani³

et al. 2007

J Ind Microbiol Biotechnol

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An alkaline protease secreting Haloalkaliphilic bacterium (Gene bank accession number EU118361) was isolated from the Saurashtra Coast in Western India. The alkaline protease was purified by a single step chromatography on phenyl sepharose 6 FF with 28% yield. The molecular mass was 40 kDa as judged by SDS-PAGE. The enzyme displayed catalysis and stability over pH 8-13, optimally at 9-11. It was stable with 0-4 M NaCl and required 150 mM NaCl for optimum catalysis at 37 degrees C; however, the salt requirement for optimal catalysis increased with temperature. While crude enzyme was active at 25-80 degrees C (optimum at 50 degrees C), the purified enzyme had temperature optimum at 37 degrees C, which shifted to 80 degrees C in the presence of 2 M NaCl. The NaCl not only shifted the temperature profile but also enhanced the substrate affinity of the enzyme as reflected by the increase in the catalytic constant (K(cat)). The enzyme was also calcium dependent and with 2 mM Ca(+2), the activity reached to maximum at 50 degrees C. The crude enzyme was highly thermostable (37-90 degrees C); however, the purified enzyme lost its stability above 50 degrees C and its half life was enhanced by 30 and sevenfold at 60 degrees C with 1 M NaCl and 50 mM Ca(+2), respectively. The activity of the enzyme was inhibited by PMSF, indicating its serine type. While the activity was slightly enhanced by Tween-80 (0.2%) and Triton X-100 (0.05%), it marginally decreased with SDS. In addition, the enzyme was highly stable with oxidizing-reducing agents and commercial detergents and was affected by metal ions to varying extent. The study assumes significance due to the enzyme stability under the dual extremities of pH and salt coupled with moderate thermal tolerance. Besides, the facts emerged on the enzyme stability would add to the limited information on this enzyme from Haloalkaliphilic bacteria.

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Production and properties of an alkaline protease by Aureobasidium pullulans

Cited by 44 publications

References 25 publications

Purification and partial characterization of a detergent and oxidizing agent stable alkaline protease from a newly isolated Bacillus subtilis VSG-4 of tropical soil

Purification and partial characterization of a detergent and oxidizing agent stable alkaline protease from a newly isolated Bacillus subtilis VSG-4 of tropical soil

Purification and Characterization of a Novel Collagenase from Bacillus pumilus Col-J

Purification and stability characteristics of an alkaline serine protease from a newly isolated Haloalkaliphilic bacterium sp. AH-6

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