PURIFICATION AND CHARACTERIZATION OF THE NEWLY THERMOSTABLE PROTEASE PRODUCED BY Brevibacillus thermoruber LII ISOLATED FROM PADANG CERMIN HOTSPRING, INDONESIA

Zilda, Dewi Zeswita; Harmayani, Eni; Widada, Jaka; Asmara, Widya; Irianto, Hari Eko; Patantis, Gintung; Fawzya, Yusro Nuri

doi:10.15578/squalen.v9i1.91

Cited by 7 publications

(12 citation statements)

References 28 publications

(27 reference statements)

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“…Monomeric proteases with molecular weights between 60 and 66 kDa from Brevibacillus sp. have been reported in literature [ 45 , 46 , 47 ].…”

Section: Resultsmentioning

confidence: 99%

“…Weak activities were still observed at pH 5.0 and 11.0. The majority of extracellular proteases reported in literature from Brevibacillus members were alkaline proteases with optimal pH around 8.0 [ 46 , 47 , 50 , 51 ], and acid proteases were rarely reported [ 45 ].…”

Section: Resultsmentioning

confidence: 99%

“…The same inhibitory effect of heavy metal ions was observed on a thermostable protease of several Brevibacillus species and is probably related to a reaction with the protein thiol groups (converting them to mercaptides), as well as with histidine and tryptophan residues. Metal ions are potent inhibitors of protein folding [ 46 , 47 , 51 ]. Protease 32-F38 activity was not affected by a 1% concentration of methanol, ethanol, or acetone.…”

Section: Resultsmentioning

confidence: 99%

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Production and Characterization of an Extracellular Acid Protease from Thermophilic Brevibacillus sp. OA30 Isolated from an Algerian Hot Spring

Gomri¹,

Rico-Díaz

Escuder-Rodríguez

et al. 2018

Microorganisms

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Proteases have numerous biotechnological applications and the bioprospection for newly-thermostable proteases from the great biodiversity of thermophilic microorganisms inhabiting hot environments, such as geothermal sources, aims to discover more effective enzymes for processes at higher temperatures. We report in this paper the production and the characterization of a purified acid protease from strain OA30, a moderate thermophilic bacterium isolated from an Algerian hot spring. Phenotypic and genotypic study of strain OA30 was followed by the production of the extracellular protease in a physiologically-optimized medium. Strain OA30 showed multiple extracellular proteolytic enzymes and protease 32-F38 was purified by chromatographic methods and its biochemical characteristics were studied. Strain OA30 was affiliated with Brevibacillus thermoruber species. Protease 32-F38 had an estimated molecular weight of 64.6 kDa and was optimally active at 50 °C. It showed a great thermostability after 240 min and its optimum pH was 6.0. Protease 32-F38 was highly stable in the presence of different detergents and solvents and was inhibited by metalloprotease inhibitors. The results of this work suggest that protease 32-F38 might have interesting biotechnological applications.

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“…Monomeric proteases with molecular weights between 60 and 66 kDa from Brevibacillus sp. have been reported in literature [ 45 , 46 , 47 ].…”

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Production and Characterization of an Extracellular Acid Protease from Thermophilic Brevibacillus sp. OA30 Isolated from an Algerian Hot Spring

Gomri¹,

Rico-Díaz

Escuder-Rodríguez

et al. 2018

Microorganisms

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show abstract

“…The concentrated enzyme precipitate was separated by centrifugation (EDLAB Clinical, Indonesia) at 15,000 rpm for 10 mins. (Zilda et al, 2014) . The precipitate obtained was then dialyzed by placing it in a cellophane bag, then soaking it in 0.05 and 0.025 M phosphate bu ers at pH 7 and stirring using a stirrer at 4°C for one night.…”

Section: Partial Puri Cation Of Bacterial Proteasementioning

confidence: 99%

Characteristics and Substrate Specificity of Semi-Purified Bacterial Protease ofBacillusthuringiensisHSFI-12 with Potential as Antithrombotic Agent

Ferdiani

Zilda²,

Afriansyah

et al. 2023

Sci. Technol. Indones

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Commercial proteases, such as Nattokinase (NK), Staphylokinase (SAK), and Streptokinase (SK) play an important role in the destruction of thrombus, the main cause of death in cardiovascular disease. The latest technology combining enzymes with certain drugs is the target of new research in the thrombolytic area. The first step is to develop protease from Bacillus thuringiensis HSFI-12 bacteria as an antithrombotic agent, characterization of the bacterial enzyme is necessary. This study aims to determine the specificity of protease from Bacillus thuringiensis HSFI-12 to explore its potential as an antithrombotic agent in terms of anticoagulant and fibrinolytic activities. The molecular weight and specificity of bacterial protease were determined with a zymographic method with casein as substrate. Bacillus thuringiensis HSFI-12 was first cultured on Nutrient Agar (NA) media and then on Skim Milk Agar (SMA) media. The obtained crude protease from Skim Milk Broth (SMB) was then concentrated as dialysate. Both crude and dialysate proteases were tested for their specific activity, as well as anticoagulant and fibrinolytic activities. Next, the dialysate’s molecular weight and specificity on the casein substrate were investigated using the zymographic method. As result, protease activity in crude form is lower than that in dialysate, which was 0.5570 ± 0,004 U/mL and 2.1767 ± 0,005 U/mL, respectively. The molecular weight of the obtained bacterial protease was between 117 – 133 kDa and the enzyme is capable of degrading casein as shown on the zymogram. Overall, both crude and dialysate proteases of Bacillus thuringiensis HSFI-12 show potential as an antithrombotic agent for exhibiting anticoagulant and antiplatelet activities. Yet, it could not exhibit direct fibrinolytic activity implying the possibility that the enzyme plays a role as a plasminogen activator, which can dissolve fibrin by activating plasmin.

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“…Microbial transglutaminase is not influenced by Ca 2+ , so the presence of metal chelating compounds such as EDTA could not inhibit enzyme activity (Lin et al, 2008). The activity of the purified transglutaminase was inhibited partially by PMSF indicating that active site contained serin (Susanti, 2003;Zilda, 2013). Tsai et al (1996) reported that transglutaminase produced by Streptoverticillium ladakanum is strongly inhibited by PMSF but not affected by EDTA.…”

Section: Effect Of Different Metal Ions and Inhibitorsmentioning

confidence: 99%

Purification and Characterization of Transglutaminase from Streptomyces sp. TTA 02 SDS 14

Nur’amaliyah

Zilda²,

Mubarik

2016

Squalen Bull. Marine Fisheries Postharvest Biotech.

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Streptomyces sp. TTA 02 SDS 14 is a transglutaminase producing bacteria which previously had been screened along with more than one hundred isolates. This research aimed to purify and characterize transglutaminase from this strain. Transglutaminase was purified from crude enzyme by ultrafiltration, Q-Sepharose ion exchange chromatography and Sepacryl S200 size exclusion chromatography sequentially, obtaining yield and purification fold of 1.36% and 27 folds, respectively. The molecular weight of the purified transglutaminase was 72 kDa detected by zymogram gel electrophoresis. The optimum temperature and pH were 50°C and 6. The transglutaminase was stable at 45°C and could be activated in the presence of 5 mM and 10 mM of Na+, K+, Li+,Ca2+, Mg2+, BPB (4-bromo-phenacyl bromide), and IAA (iodo acetamide acid), but the activity was inhibited by the presence of Cu+, Zn2+, and PMSF (phenyl methyl sulfonyl fluoride).

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PURIFICATION AND CHARACTERIZATION OF THE NEWLY THERMOSTABLE PROTEASE PRODUCED BY Brevibacillus thermoruber LII ISOLATED FROM PADANG CERMIN HOTSPRING, INDONESIA

Cited by 7 publications

References 28 publications

Production and Characterization of an Extracellular Acid Protease from Thermophilic Brevibacillus sp. OA30 Isolated from an Algerian Hot Spring

Production and Characterization of an Extracellular Acid Protease from Thermophilic Brevibacillus sp. OA30 Isolated from an Algerian Hot Spring

Characteristics and Substrate Specificity of Semi-Purified Bacterial Protease ofBacillusthuringiensisHSFI-12 with Potential as Antithrombotic Agent

Purification and Characterization of Transglutaminase from Streptomyces sp. TTA 02 SDS 14

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