Isolation and characterization of a new cold-active protease from psychrotrophic bacteria of Western Himalayan glacial soil

Saleem, Farooq; Nazir, Ruqeya; Ganai, Shabir Ahmad; Ganai, Bashir Ahmad

doi:10.1038/s41598-021-92197-w

Cited by 25 publications

(13 citation statements)

References 85 publications

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“…No effect on the activity of a serine protease from B. clausii GMBAE 42 after incubation with EDTA and a slight inhibition by Ca 2+ was also found by Kazan et al [49]. Others could show a strong activity decrease after the incubation with EDTA [58,79,80]. However, most studies found a comparable decline in activity as seen in this work [46,81,82].…”

Section: Effect Of Sds and H 2 O 2 On Enzyme Activitysupporting

confidence: 82%

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Biochemical characterization of a novel oxidatively stable, halotolerant, and high‐alkaline subtilisin from Alkalihalobacillus okhensisKh10‐101^T

et al. 2022

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Halophilic and halotolerant microorganisms represent a promising source of salt‐tolerant enzymes suitable for various biotechnological applications where high salt concentrations would otherwise limit enzymatic activity. Considering the current growing enzyme market and the need for more efficient and new biocatalysts, the present study aimed at the characterization of a high‐alkaline subtilisin from Alkalihalobacillus okhensis Kh10‐101 T . The protease gene was cloned and expressed in Bacillus subtilis DB104. The recombinant protease SPAO with 269 amino acids belongs to the subfamily of high‐alkaline subtilisins. The biochemical characteristics of purified SPAO were analyzed in comparison with subtilisin Carlsberg, Savinase, and BPN'. SPAO, a monomer with a molecular mass of 27.1 kDa, was active over a wide range of pH 6.0–12.0 and temperature 20–80 °C, optimally at pH 9.0–9.5 and 55 °C. The protease is highly oxidatively stable to hydrogen peroxide and retained 58% of residual activity when incubated at 10 °C with 5% (v/v) H 2 O 2 for 1 h while stimulated at 1% (v/v) H 2 O 2 . Furthermore, SPAO was very stable and active at NaCl concentrations up to 5.0 m . This study demonstrates the potential of SPAO for biotechnological applications in the future.

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Section: Effect Of Sds and H 2 O 2 On Enzyme Activitysupporting

confidence: 82%

“…[ 49 ]. Others could show a strong activity decrease after the incubation with EDTA [ 58 , 79 , 80 ]. However, most studies found a comparable decline in activity as seen in this work [ 46 , 81 , 82 ].…”

Section: Resultsmentioning

confidence: 99%

Biochemical characterization of a novel oxidatively stable, halotolerant, and high‐alkaline subtilisin from Alkalihalobacillus okhensisKh10‐101^T

et al. 2022

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“…Bacillus sp. HM49 was observed to yield maximum protease enzyme by incubating at 20 °C which was in accordance with a study on Bacillus pumilus isolated from glacial soils of Thajwas Glacier [ 57 ]. However, Nörnberg et al [ 58 ] reported protease production from psychrotrophic isolates at 30 °C.…”

Section: Discussionsupporting

confidence: 89%

Molecular and functional characterization of protease from psychrotrophic Bacillus sp. HM49 in North-western Himalaya

et al. 2023

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In this work, a psychrotrophic bacteria producing cold-active protease, was obtained from Dachigam National Park, an ecologically significant habitat in Western Himalayas owing to its varied endemic and endangered flora and fauna. This isolate was identified as Bacillus sp. HM49 via phenotypic, Gram staining, bio-chemical and 16S rRNA gene identification. Isolate HM49 when tested for proteolytic activity revealed prominent hydrolytic zone with the most production at 20 °C and pH, 8.0 post 72 h incubation. This enzyme was purified, enhancing its specific activity to 61.15 U/mg and its characterization studies revealed it to be a cold-alkaline protease being active in a wide pH (6.0–12) and temperature (5–40 °C) range. Amplification of CAASPR gene of HM49 was performed, followed by enzyme-substrate docking studies and MMGBSA providing details about its type, molecular weight validation as well as functional applications. The purified protease of HM49 was tested for laundry applications and the enzyme was found to be compatible with majority of the detergents tested. Its potential as an eco-friendly detergent additive was further validated by wash performance test as it effectively removed recalcitrant blood stains at a low temperature of 20 °C that could be beneficial for fine garments like silk which preferably need cold washing.

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“…This pH dependency for activity makes the isolated protease remarkable for industrial uses, such as additives in washing detergents, leather processing, or other applications in comparison to other cold proteases with a narrow pH profile [38]. The detergent‐stable proteases available commercially like subtilisin, and savinase are also stable at high pH (≥9) [36, 39]. The washing effectiveness of such cold‐active proteases at a wide range of pH and with stability to detergents at normal temperature makes it more appreciated.…”

Section: Discussionmentioning

confidence: 99%

Isolation and characterization of a cold‐active, detergent‐stable protease from Serratia sp. TGS1

et al. 2023

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Psychrophiles are cold‐adapted microorganisms living in cold regions and are known to generate cold‐active enzymes such as proteases, lipases, and peptidases. These types of enzymes are a major part of the market of the food and textile sector. This study aimed to isolate and characterize the cold‐active and detergent‐stable, extracellular protease from psychotrophic bacteria Serratia sp. TGS1 (OQ654005). Protease was purified by gel permeation chromatography using Sephadex G‐75. The specific activity of the purified protease was 250 U/mg at 15°C, with a purification fold of 5.68 and a percentage yield of 60%. The cold active protease was stable within a temperature range of 5–30°C and a pH range of 6–10. Ca+2 and Mg+2 enhanced its activity while chelators like ethylenediaminetetraacetic acid inhibited cold active protease, showing it as metalloprotease in nature. The enzyme was sensitive to Cu+2, Zn+2, and Hg+2, and the proteolytic activity decreased upon treatment with heavy metals. The molecular weight of the protease was estimated to be 47 kDa using sodium dodecyl sulfate (SDS)‐polyacrylamide gel electrophoresis. Proteins within a specific range of molecular weight possess desirable properties for industrial enzyme use. By working on a specific range, the researchers intended to examine an enzyme to examine its specific characteristics. The purified protease showed high stability to detergents like SDS, Tween 20, Tween 60, and Triton X. The maximum velocity Vmax and Km values were 59.90 mg/min/mL and 1.53 mg/mL, respectively. The obtained protease exhibited an interesting activity at a broad range of pH (6–10) and stability at low temperatures (5–30°C) and detergents. Such enzymatic features of versatile and potent cold‐active enzymes enhance their industrial applications to meet food, dairy, and laundry requirements.

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Isolation and characterization of a new cold-active protease from psychrotrophic bacteria of Western Himalayan glacial soil

Cited by 25 publications

References 85 publications

Biochemical characterization of a novel oxidatively stable, halotolerant, and high‐alkaline subtilisin from Alkalihalobacillus okhensisKh10‐101^T

Biochemical characterization of a novel oxidatively stable, halotolerant, and high‐alkaline subtilisin from Alkalihalobacillus okhensisKh10‐101^T

Molecular and functional characterization of protease from psychrotrophic Bacillus sp. HM49 in North-western Himalaya

Isolation and characterization of a cold‐active, detergent‐stable protease from Serratia sp. TGS1

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Isolation and characterization of a new cold-active protease from psychrotrophic bacteria of Western Himalayan glacial soil

Cited by 25 publications

References 85 publications

Biochemical characterization of a novel oxidatively stable, halotolerant, and high‐alkaline subtilisin from Alkalihalobacillus okhensisKh10‐101T

Biochemical characterization of a novel oxidatively stable, halotolerant, and high‐alkaline subtilisin from Alkalihalobacillus okhensisKh10‐101T

Molecular and functional characterization of protease from psychrotrophic Bacillus sp. HM49 in North-western Himalaya

Isolation and characterization of a cold‐active, detergent‐stable protease from Serratia sp. TGS1

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Biochemical characterization of a novel oxidatively stable, halotolerant, and high‐alkaline subtilisin from Alkalihalobacillus okhensisKh10‐101^T

Biochemical characterization of a novel oxidatively stable, halotolerant, and high‐alkaline subtilisin from Alkalihalobacillus okhensisKh10‐101^T