Production of detergent stable thermophilic alkaline protease by Bacillus licheniformis ALW1

Emran, Mohamed A.; Ismail, S. I.; Hashem, Ahmed M.

doi:10.1016/j.bcab.2020.101631

Cited by 36 publications

(15 citation statements)

References 35 publications

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“…These values mostly rely on the enzyme source (microorganism) and the type of substrate, thereby indicating the sensitive nature of the enzyme against its substrate, which is believed to increase as the K M value increases while the V max value decreases [ 100 ]. Values of K M (11.71 mg mL −1 ) and V max (357.14 µmol mL −1 min −1 ) were computed with good correlation (R2 = 0.9986), which was higher than many other previously reported values of K M and V max for several bacterial species as 3.846 mg mL −1 and 76.923 U mL −1 min −1 [ 101 ]; 3.99 mg mL −1 and 41.49 U mL −1 [ 102 ].…”

Section: Discussionmentioning

confidence: 58%

Biochemical Characterization and Functional Analysis of Heat Stable High Potential Protease of Bacillus amyloliquefaciens Strain HM48 from Soils of Dachigam National Park in Kashmir Himalaya

et al. 2021

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A novel temperature stable alkaline protease yielding bacteria was isolated from the soils of Dachigam National Park, which is known to be inhabited by a wide variety of endemic plant and animal species of Western Himalaya. This high-potential protease producing isolate was characterized and identified as Bacillus amyloliquefaciens strain HM48 by morphological, Gram’s staining and biochemical techniques followed by molecular characterization using 16S rRNA approach. The extracellular protease of B. amyloliquefaciens HM48 was purified by precipitating with ammonium sulfate (80%), followed by dialysis and Gel filtration chromatography increasing its purity by 5.8-fold. The SDS–PAGE analysis of the purified enzyme confirmed a molecular weight of about ≈25 kDa. The enzyme displayed exceptional activity in a broad temperature range (10–90 °C) at pH 8.0, retaining its maximum at 70 °C, being the highest reported for this proteolytic Bacillus sp., with KM and Vmax of 11.71 mg/mL and 357.14 µmol/mL/min, respectively. The enzyme exhibited remarkable activity and stability against various metal ions, surfactants, oxidizing agent (H2O2), organic solvents and displayed outstanding compatibility with widely used detergents. This protease showed effective wash performance by exemplifying complete blood and egg-yolk stains removal at 70 °C and efficiently disintegrated chicken feathers making it of vital importance for laundry purpose and waste management. For functional analysis, protease gene amplification of strain HM48 yielded a nucleotide sequence of about 700 bp, which, when checked against the available sequences in NCBI, displayed similarity with subtilisin-like serine protease of B. amyloliquefaciens. The structure of this protease and its highest-priority substrate β-casein was generated through protein modeling. These protein models were validated through futuristic algorithms following which protein–protein (protease from HM48 and β-casein) docking was performed. The interaction profile of these proteins in the docked state with each other was also generated, shedding light on their finer details. Such attributes make this thermally stable protease novel and suitable for high-temperature industrial and environmental applications.

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Section: Discussionmentioning

confidence: 58%

Biochemical Characterization and Functional Analysis of Heat Stable High Potential Protease of Bacillus amyloliquefaciens Strain HM48 from Soils of Dachigam National Park in Kashmir Himalaya

et al. 2021

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“…For example, alkaline protease was prepared from Bacillus licheniformis ALW1 with the productivity of 22.90 U mL −1 . 22 Besides, a novel extracellular alkaline protease called SAPHM was prepared from Bacillus licheniformis strain K7A, optimally active at pH 10 and 70 °C. 23 In addition, chitinase and ⊍-amylase have also been successfully prepared from Bacillus licheniformis, 24,25 demonstrating Bacillus licheniformis and its products can be widely used in many fields related to chemistry, medicine, food and feed.…”

Section: Resultsmentioning

confidence: 99%

“…At last, a phylogenetic tree was constructed using MEGA 7.0. 15 Growth curve and microbial growth kinetics After inoculation with 2 mL of the earlier seed culture into LB liquid medium (50 mL) contained in 66 shake flasks (250 mL) marked from A 1 to A 22 , B 1 to B 22 and C 1 to C 22 , respectively, the mixtures were incubated at 60 °C and 160 rpm, and their absorbance values were determined at 600 nm at intervals of 30 min (0-5 h), 1 h (5-8 h) and 2 h (8-24 h) and the growth curve was plotted accordingly. Strain incubated at 30 °C for a total period of 16 h was used as a control.…”

Section: Determination Of Protein Contents and Trypsin Inhibitor Acti...mentioning

confidence: 99%

Identification of a thermophilic protease‐producing strain and its application in solid‐state fermentation of soybean meal

Dai

Hou

et al. 2021

J Sci Food Agric

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BACKGROUND: Thermophiles can thrive at 50-80 °C and produce some enzymes with special promise for biocatalysis. A thermophilic protease-producing strain YYC4 was isolated from Yunyan cigarette and employed in solid-state fermentation (SSF) of unsterilized soybean meal (SBM). RESULTS:The isolate was identified as Bacillus licheniformis based on appearance of colonies, microscopic observation and 16S rDNA sequencing. After SSF, soluble and crude protein contents in SBM increased from 49.24 to 185.73 g kg −1 and from 404.18 to 479.46 g kg −1 , respectively, under the fermentation conditions of 10 7 cfu g −1 inoculation of strain YYC4, 1:1.8 (g mL −1 ) SBM to distilled water, 1.2 g kg −1 magnesium sulphate addition, 55 °C and 48 h. During fermentation, pH of the medium increased from 6.30 to 9.09 and protease activity especially neutral protease increased significantly from 13.5 to 181.31 U g −1 . Meanwhile, trypsin inhibitor (TI) activity was decreased from 8.19 to 3.19 mg g −1 . The safety of fermented SBM (FSBM) was verified by acute toxicity animal experiment. Analysis of microbial community in FSBM showed that Bacillus licheniformis YYC4 as a dominant strain inhibited most of the other microorganisms pre-existing in the materials during fermentation.CONCLUSION: Increments of soluble and crude protein by 277.19% and 18.63% and decrement of harmful TI by 61.05% in SBM were achieved using thermophilic SSF by Bacillus licheniformis YYC4, providing a basis for the application of thermophiles in fermentation industry in an environmentally friendly and energy-saving way.

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“…For instance, thermophilic proteases can be used as a protein degrader in waste treatment processes, e.g., the solubilization of sewage sludge ( Nakamichi et al, 2010 ; Kun et al, 2018 ). They also can be used as the detergents in high-temperature conditions ( Mechri et al, 2019 ; Emran et al, 2020 ). The results in this study indicate that Anoxybacillus strain 1A02591 is likely to be developed as a good producer of thermophilic proteases that may have various biotechnological applications.…”

Section: Discussionmentioning

confidence: 99%

Characterization and Diversity Analysis of the Extracellular Proteases of Thermophilic Anoxybacillus caldiproteolyticus 1A02591 From Deep-Sea Hydrothermal Vent Sediment

Cheng

Wang²,

Zhang³

et al. 2021

Front. Microbiol.

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Protease-producing bacteria play key roles in the degradation of marine organic nitrogen. Although some deep-sea bacteria are found to produce proteases, there has been no report on protease-secreting Anoxybacillus from marine hydrothermal vent regions. Here, we analyzed the diversity and functions of the proteases, especially the extracellular proteases, of Anoxybacillus caldiproteolyticus 1A02591, a protease-secreting strain isolated from a deep-sea hydrothermal vent sediment of the East Pacific Ocean. Strain 1A02591 is a thermophilic bacterium with a strong protease-secreting ability, which displayed the maximum growth rate (0.139 h–1) and extracellular protease production (307.99 U/mL) at 55°C. Strain 1A02591 contains 75 putative proteases, including 65 intracellular proteases and 10 extracellular proteases according to signal peptide prediction. When strain 1A02591 was cultured with casein, 12 proteases were identified in the secretome, in which metalloproteases (6/12) and serine proteases (4/12) accounted for the majority, and a thermolysin-like protease of the M4 family was the most abundant, suggesting that strain 1A02591 mainly secreted a thermophilic metalloprotease. Correspondingly, the secreted proteases of strain 1A02591 showed the highest activity at the temperature as high as 70°C, and was inhibited 70% by metalloprotease inhibitor o-phenanthroline and 50% by serine protease inhibitor phenylmethylsulfonyl fluoride. The secreted proteases could degrade different proteins, suggesting the role of strain 1A02591 in organic nitrogen degradation in deep-sea hydrothermal ecosystem. These results provide the first insight into the proteases of an Anoxybacillus strain from deep-sea hydrothermal ecosystem, which is helpful in understanding the function of Anoxybacillus in the marine biogeochemical cycle.

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Production of detergent stable thermophilic alkaline protease by Bacillus licheniformis ALW1

Cited by 36 publications

References 35 publications

Biochemical Characterization and Functional Analysis of Heat Stable High Potential Protease of Bacillus amyloliquefaciens Strain HM48 from Soils of Dachigam National Park in Kashmir Himalaya

Biochemical Characterization and Functional Analysis of Heat Stable High Potential Protease of Bacillus amyloliquefaciens Strain HM48 from Soils of Dachigam National Park in Kashmir Himalaya

Identification of a thermophilic protease‐producing strain and its application in solid‐state fermentation of soybean meal

Characterization and Diversity Analysis of the Extracellular Proteases of Thermophilic Anoxybacillus caldiproteolyticus 1A02591 From Deep-Sea Hydrothermal Vent Sediment

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