scholarly journals Effect of Cultural Conditions on Protease Production by a Thermophilic Geobacillus thermoglucosidasius SKF4 Isolated from Sungai Klah Hot Spring Park, Malaysia

Molecules ◽  
2020 ◽  
Vol 25 (11) ◽  
pp. 2609
Author(s):  
Allison D. Suleiman ◽  
Nor’Aini Abdul Rahman ◽  
Hidayat Mohd Yusof ◽  
Fairolniza Mohd Shariff ◽  
Nur Adeela Yasid
Keyword(s):  
Incubation Time ◽  
Hot Spring ◽  
Nitrogen Sources ◽  
Protease Production ◽  
Rrna Gene ◽  
16S Rrna Gene Analysis ◽  
Geobacillus Thermoglucosidasius ◽  
Cultural Conditions ◽  
Protease Enzyme ◽  
Nacl Concentration

Major progress in the fields of agriculture, industry, and biotechnology over the years has influenced the quest for a potent microorganism with favorable properties to be used in scientific research and industry. This study intended to isolate a new thermophilic-protease-producing bacterium and evaluate its growth and protease production under cultural conditions. Protease producing bacteria were successfully isolated from Sungai Klah Hot Spring Park in Perak, Malaysia, and coded as SKF4; they were promising protease producers. Based on microscopic, morphological, and 16S rRNA gene analysis, isolate SKF4 was identified as Geobacillus thermoglucosidasius SKF4. The process of isolating SKF4 to grow and produce proteases under different cultural conditions, including temperature, pH, NaCl concentration, carbon and nitrogen sources, and incubation time, was explored. The optimum cultural conditions observed for growth and protease production were at 60 to 65 °C of temperature, pH 7 to 8, and under 1% NaCl concentration. Further, the use of casein and yeast extract as the nitrogen sources, and sucrose and fructose as the carbon sources enhanced the growth and protease production of isolate SKF4. Meanwhile, isolate SKF4 reached maximum growth and protease production at 24 h of incubation time. The results of this study revealed a new potent strain of thermophilic bacterium isolated from Sungai Klah Hot Spring Park in Perak, Malaysia for the first time. The high production of thermostable protease enzyme by G. thermoglucosidasius SKF4 highlighted the promising properties of this bacterium for industrial and biotechnological applications.

scholarly journals Keratinolytic Protease Production by Bacillus Cereus Strain Ps03 under Submerged Fermentation: Optimization and Characterization

2016 ◽  
Vol 4 (3) ◽  
pp. 397-401
Author(s):  
M.D. BalaKumaran ◽  
R. Santhi
Keyword(s):  
Bacillus Subtilis ◽  
Scale Up ◽  
Nitrogen Sources ◽  
Maximum Activity ◽  
Protease Production ◽  
Subtilis Strain ◽  
Physical Factors ◽  
Chicken Feather ◽  
Protease Enzyme ◽  
Keratinolytic Protease

In the present study, chicken feather powder was screened for its application as the substrate for the production of keratinolytic protease by Bacillus subtilis strain PS03. Bacillus subtilis produced a high level of keratinolytic protease using chicken feather powder as substrate. With feather powder as substrate, physical factors such as incubation time, pH and temperature were optimized for increased keratinolytic protease production by Bacillus subtilis. The enzyme production was enhanced when using maltose as carbon source and yeast extract as nitrogen sources. SDS-PAGE analysis indicated the molecular weight of 46 kDa of the partially purified keratinolytic protease. The keratinolytic protease enzyme was stable over a pH range of 6 – 9 and temperature range of 35 - 50°C with maximum activity at pH 9 and 40°C. Based on the results, the use of feather powder as substrate for keratinolytic protease production is cost effective and is easy to scale up. Considering the availability and cost, chicken feather powder is considered as an ideal substrate for keratinolytic protease production in an industrial point of view. Int J Appl Sci Biotechnol, Vol 4(3): 397-401

scholarly journals A Novel Thermostable and Alkaline Protease Produced from Bacillus stearothermophilus Isolated from Olive Oil Mill Sols Suitable to Industrial Biotechnology

Molecules ◽  
2021 ◽  
Vol 26 (4) ◽  
pp. 1139
Author(s):  
Aida Karray ◽  
Mona Alonazi ◽  
Habib Horchani ◽  
Abir Ben Bacha
Keyword(s):  
Cell Growth ◽  
Olive Oil ◽  
Incubation Time ◽  
Alkaline Protease ◽  
Bacillus Stearothermophilus ◽  
Nitrogen Sources ◽  
Reversed Phase ◽  
Culture Conditions ◽  
Protease Production ◽  
Industrial Biotechnology

This study was conducted to identify a new alkaline and thermophilic protease (Ba.St.Pr) produced from Bacillus stearothermophilus isolated from olive oil mill sols and to evaluate its culture conditions, including temperature, pH, carbon and nitrogen sources, and incubation time. The optimum culture conditions for cell growth (10 g/L) and protease production (5050 U/mL) were as follows: temperature 55 °C, pH 10, inoculation density 8 × 108 CFU/mL, and incubation time 24 h. The use of 3% yeast extract as the nitrogen sources and galactose (7.5 g/L) as the carbon sources enhanced both cell growth and protease production. Using reversed-phase analytical HPLC on C-8 column, the new protease was purified with a molecular mass of approximately 28 kDa. The N-terminal sequence of Ba.St.Pr exhibited a high level of identity of approximately 95% with those of Bacillus strains. Characterization under extreme conditions revealed a novel thermostable and alkaline protease with a half-life time of 187 min when incubated with combined Ca2+/mannitol. Ba.St.Pr demonstrated a higher stability in the presence of surfactant, solvent, and Ca2+ ions. Consequently, all the evaluated activity parameters highlighted the promising properties of this bacterium for industrial and biotechnological applications.

scholarly journals CULTIVATION CONDITIONS FOR PROTEASE PRODUCTION BY A THERMO-HALOSTABLE BACTERIAL ISOLATE PLS A

2019 ◽  
Vol 19 (1) ◽  
pp. 18-23
Author(s):  
Teuku M. Iqbalsyah ◽  
Malahayati Malahayati ◽  
Atikah Atikah ◽  
Febriani Febriani
Keyword(s):  
Protease Activity ◽  
Hot Springs ◽  
Skim Milk ◽  
Dry Weight ◽  
Protease Production ◽  
Cultivation Conditions ◽  
Thermostable Enzymes ◽  
Cultivation Period ◽  
Protease Enzyme ◽  
Nacl Concentration

Polyextremophiles have increasingly been utilised to produce thermostable enzymes with better stability in multiple extreme conditions. This study reports the screening results of four new bacterial isolates (PLS A, PLS 75, PLS 76 and PLS 80), isolated from an under water hot springs, in producing thermo-halostable protease enzyme. Optimum cultivation conditions for the protease production were also studied. Screening of protease-producing isolates was conducted using Thermus solid medium enriched with 3% skim milk and 0.5% casein. The growth of the isolates showing protease activity was monitored by measuring the cell dry weight and protease activity during 24 h cultivation period. The activity was also measured at various cultivation conditions, i.e. temperature, pH and salt concentrations. Amongst the four isolates, only PLS A showed the ability to produce protease. The optimum cultivation conditions for protease production were observed at 65°C, pH 7 for 18 h incubation. The activity increased with the addition of 1% NaCl concentration (0.085 Unit/mL). The ability of PLS A isolate to produce thermo-halostable protease was encouraging as they could potentially be used in industries requiring the enzyme with multiple extremes. 

scholarly journals Protease Production from UV Mutated Bacillus subtilis

2012 ◽  
Vol 47 (1) ◽  
pp. 69-76 ◽  
Author(s):  
MG Sher ◽  
M Nadeem ◽  
Q Syed ◽  
M Irfan ◽  
S Baig
Keyword(s):  
Bacillus Subtilis ◽  
Inorganic Nitrogen ◽  
Carbon Sources ◽  
Nitrogen Sources ◽  
Fermentation Medium ◽  
Maximum Activity ◽  
Inoculum Size ◽  
Protease Production ◽  
Protease Enzyme ◽  
Uv Mutation

UV mutation of the strain has significant contributation to enhance the yield of protease enzyme from Bacillus subtilis bacteria under the  cultivation conditions in submerged fermentation. The fermentation medium used for the production of protease composed of carbon  sources 1%, organic 1% or inorganic nitrogen sources 0.5% , K2HPO4 0.2 %, CaCl2 0.04% and MgSO4 0.02 % by mutated Bacillus subtilis  G-4 under the optimum parameters which are important to induce the mutated strain to produce high units of the protease, which were  temperature 37.5°C, pH 9, inoculum size 3 % v/v, glucose 1% as carbon source and peptone 1% as nitrogen source were give the maximum  455.25 + 1.66 units of protease. The results of stability studies revealed that protease of B. subtilis G-4 was stable over a broad range  of temperature (30 to 60°C) and pH (8 to 12). However, maximum activity (155.45U/ml) was observed at temperature 50°C and pH 10.  These characteristics render its potential use in detergent industries for detergent formulation.DOI: http://dx.doi.org/10.3329/bjsir.v47i1.10725 Bangladesh J. Sci. Ind. Res. 47(1), 69-76, 2012

scholarly journals Screening of Nutritional Parameters for the Production of Protease fromAspergillus Oryzae

2007 ◽  
Vol 4 (2) ◽  
pp. 208-215 ◽  
Author(s):  
G. Srinubabu ◽  
N. Lokeswari ◽  
K. Jayaraju
Keyword(s):  
Carbon Sources ◽  
Nitrogen Sources ◽  
Fold Increase ◽  
Protease Production ◽  
Malt Extract ◽  
Culture Time ◽  
Protease Enzyme ◽  
Bean Meal ◽  
Soya Bean Meal ◽  
Fungus Growth

Production of protease enzyme by fungusAspergillus oryzaewas investigated. The proteolytic activity was observed when the fungus was grown in the medium containing glucose, malt extract, yeast extract, peptone, K2HPO4, MgSO4and FeSO4. The present paper describes the screening of media components and fermentation conditions in shake flask. The organism utilized carbon sources glucose, fructose, sucrose, lactose, dextrin and starch among them glucose was found to be the best carbon source, for nitrogen sources various inorganic and organic media components were investigated among them peptone is found to be the best nitrogen source. 1% cottonseed followed by 2% Soya bean meal was found to be the best inducer. With optimized media two-fold increase in the protease production. The fungus growth depends on the concentration of carbon, nitrogen and salt solution, where as the enzyme production was also influenced by the culture time, pH and interaction between these two variables.

scholarly journals Metallosphaera cuprina sp. nov., an acidothermophilic, metal-mobilizing archaeon

2011 ◽  
Vol 61 (10) ◽  
pp. 2395-2400 ◽  
Author(s):  
Li-Jun Liu ◽  
Xiao-Yan You ◽  
Xu Guo ◽  
Shuang-Jiang Liu ◽  
Cheng-Ying Jiang
Keyword(s):  
Hot Spring ◽  
Carbon Sources ◽  
Rrna Gene ◽  
Reduced Sulfur Compounds ◽  
Dna Relatedness ◽  
Metallosphaera Sedula ◽  
Gram Staining ◽  
Nacl Concentration ◽  
Cellular Lipids ◽  
Ph Range

A novel acidothermophilic archaeon, strain Ar-4T, was isolated from a sulfuric hot spring in Tengchong, Yunnan, China. Cells of strain Ar-4T were Gram-staining-negative, irregular cocci and motile by means of flagella. Strain Ar-4T grew over a temperature range of 55–75 °C (optimum, 65 °C), a pH range of 2.5–5.5 (optimum, pH 3.5) and a NaCl concentration range of 0–1 % (w/v). The novel strain was aerobic and facultatively chemolithoautotrophic. The strain could extract metal ions from sulfidic ore. It was also able to oxidize reduced sulfur compounds. In addition, it was able to use heterogeneous organic materials for organotrophic growth. The main cellular lipids were calditoglycerocaldarchaeol (CGTE) and caldarchaeol (DGTE). The DNA G+C content of the strain was 40.2 mol%. Analysis of 16S rRNA gene sequences showed that strain Ar-4T was phylogenetically related to members of the genus Metallosphaera and had sequence similarities of 97.7 %, 97.0 % and 96.8 % with Metallosphaera hakonensis DSM 7519T, Metallosphaera sedula DSM 5348T and Metallosphaera prunae DSM 10039T, respectively. Strain Ar-4T showed DNA–DNA relatedness values of 47.5 %, 30.8 % and 29.1 % with M. hakonensis DSM 7519T, M. sedula DSM 5348T and M. prunae DSM 10039T, respectively. The differences in cell motility, the temperature and pH ranges for growth, the ability to utilize carbon sources, the DNA G+C content, and the low DNA–DNA relatedness values distinguished strain Ar-4T from recognized species of the genus Metallosphaera. On the basis of these results, it was concluded that strain Ar-4T represents a novel species of the genus Metallosphaera, for which the name Metallosphaera cuprina is proposed. The type strain is Ar-4T ( = JCM 15769T = CGMCC 1.7082T).

Cytotoxic Effect of Pseudomonas aurogenosa Protease on Cancer Cells

2019 ◽  
Vol 10 (03) ◽  
Author(s):  
Ghanyia J. Shanyoor ◽  
Fatima R. Abdul ◽  
Nehad A. Taher ◽  
Ihsan A. Raheem
Keyword(s):  
Cell Line ◽  
Normal Cell ◽  
Cytotoxic Effect ◽  
Human Cancer ◽  
Skim Milk ◽  
Exchange Chromatography ◽  
Protease Production ◽  
Normal Cell Line ◽  
Protease Enzyme ◽  
Embryonic Fibroblast

About (20) Pseudomonas rogenosa isolate were experienced for their ability of protease production by calculating the diameter of lysis area after developing on skim milk agar medium (qualitatively ) and the results exhibited that only isolate no (5), was higher isolate for protease making of (26mm) of lysis area. Then, the protein concentration also identified by Bradford method and it was found of 0.16 mg/ ml , then purification was done by using an ion- exchange chromatography with DEAE sephadex G- 100 column and the results showed the presence of 1 peak of enzyme with 50 Kd of molecular weight 2 peaks of other proteins . we tried to investigate the invitro Cytotoxic effect of purified enzyme against two human cancer lines, HeP2 (Human larynx epidermed carcinoma ) , RD ( Rabdo- Sarcoma ) , and one normal cell line Ref ( Rat embryonic fibroblast ) . The cancer and normal cells were treated with different concentrations of protease enzyme ranging from ( 0.05, 0.1, 0.2, 0.4,0.8and 0.16 mg/ml) then incubated for additional 48h at 37C0 and the results showed highest toxicity ( 80.28%) of protease enzyme on RD , moderate cytotoxicity (45.52%) on Hep andslight toxicity ( 37.12% ) on normal cell line (Ref) in a concentration (0.8mg/ml).

Constraints in Adoption ofModern Farm Machines by Tribal Farmers in Ramgarh District of Jharkhand

2019 ◽  
Vol 6 (03) ◽  
Author(s):  
PK SUNDARAM ◽  
BIKASH SARKAR ◽  
UJJWAL KUMAR ◽  
AP ANURAG ◽  
DK RAGHAV ◽  
...  
Keyword(s):  
Cell Line ◽  
Normal Cell ◽  
Human Cancer ◽  
Skim Milk ◽  
Exchange Chromatography ◽  
Protease Production ◽  
Normal Cell Line ◽  
Protease Enzyme ◽  
Tribal Farmers ◽  
Embryonic Fibroblast

About (20) Pseudomonas rogenosa isolate were experienced for their ability of protease production by calculating the diameter of lysis area after developing on skim milk agar medium (qualitatively ) andamp; the results exhibited that only isolate no (5), was higher isolate for protease making of (26mm) of lysis area. Then, the protein concentration also identified by Bradford method andamp; it was found of 0.16 mg/ ml , then purification was done by using an ion- exchange chromatography with DEAE sephadex G- 100 column andamp; the results showed the presence of 1 peak of enzyme with 50 Kd of molecular weight 2 peaks of other proteins . we tried to investigate the invitro Cytotoxic effect of purified enzyme against two human cancer lines, HeP2 (Human larynx epidermed carcinoma ) , RD ( Rabdo- Sarcoma ) , andamp; one normal cell line Ref ( Rat embryonic fibroblast ). The cancer andamp; normal cells were treated with different concentrations of protease enzyme ranging from ( 0.05, 0.1, 0.2, 0.4,0.8andamp; 0.16 mg/ml) then incubated for additional 48h at 37C 0 andamp; the results showed highest toxicity ( 80.28%) of protease enzyme on RD , moderate cytotoxicity (45.52%) on Hep andamp;slight toxicity ( 37.12% )on normal cell line (Ref) in a concentration (0.8mg/ml).

scholarly journals Microbiome of Odontogenic Abscesses

2021 ◽  
Vol 9 (6) ◽  
pp. 1307
Author(s):  
Sebastian Böttger ◽  
Silke Zechel-Gran ◽  
Daniel Schmermund ◽  
Philipp Streckbein ◽  
Jan-Falco Wilbrand ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Oral Microbiome ◽  
Minor Role ◽  
Rrna Gene ◽  
Sequencing Analysis ◽  
Bacterial Strains ◽  
16S Rrna Gene Analysis ◽  
Odontogenic Infections ◽  
Odontogenic Abscess

Severe odontogenic abscesses are regularly caused by bacteria of the physiological oral microbiome. However, the culture of these bacteria is often prone to errors and sometimes does not result in any bacterial growth. Furthermore, various authors found completely different bacterial spectra in odontogenic abscesses. Experimental 16S rRNA gene next-generation sequencing analysis was used to identify the microbiome of the saliva and the pus in patients with a severe odontogenic infection. The microbiome of the saliva and the pus was determined for 50 patients with a severe odontogenic abscess. Perimandibular and submandibular abscesses were the most commonly observed diseases at 15 (30%) patients each. Polymicrobial infections were observed in 48 (96%) cases, while the picture of a mono-infection only occurred twice (4%). On average, 31.44 (±12.09) bacterial genera were detected in the pus and 41.32 (±9.00) in the saliva. In most cases, a predominantly anaerobic bacterial spectrum was found in the pus, while saliva showed a similar oral microbiome to healthy individuals. In the majority of cases, odontogenic infections are polymicrobial. Our results indicate that these are mainly caused by anaerobic bacterial strains and that aerobic and facultative anaerobe bacteria seem to play a more minor role than previously described by other authors. The 16S rRNA gene analysis detects significantly more bacteria than conventional methods and molecular methods should therefore become a part of routine diagnostics in medical microbiology.

scholarly journals Microbial Diversity of Terrestrial Geothermal Springs in Armenia and Nagorno-Karabakh: A Review

2021 ◽  
Vol 9 (7) ◽  
pp. 1473
Author(s):  
Ani Saghatelyan ◽  
Armine Margaryan ◽  
Hovik Panosyan ◽  
Nils-Kåre Birkeland
Keyword(s):  
16S Rrna ◽  
Microbial Diversity ◽  
High Altitude ◽  
Hot Springs ◽  
Hot Spring ◽  
Abiotic Factors ◽  
Rrna Gene ◽  
Geothermal Springs ◽  
Bacterial Phyla ◽  
Terrestrial Hot Springs

The microbial diversity of high-altitude geothermal springs has been recently assessed to explore their biotechnological potential. However, little is known regarding the microbiota of similar ecosystems located on the Armenian Highland. This review summarizes the known information on the microbiota of nine high-altitude mineralized geothermal springs (temperature range 25.8–70 °C and pH range 6.0–7.5) in Armenia and Nagorno-Karabakh. All these geothermal springs are at altitudes ranging from 960–2090 m above sea level and are located on the Alpide (Alpine–Himalayan) orogenic belt, a seismically active region. A mixed-cation mixed-anion composition, with total mineralization of 0.5 mg/L, has been identified for these thermal springs. The taxonomic diversity of hot spring microbiomes has been examined using culture-independent approaches, including denaturing gradient gel electrophoresis (DGGE), 16S rRNA gene library construction, 454 pyrosequencing, and Illumina HiSeq. The bacterial phyla Proteobacteria, Bacteroidetes, Cyanobacteria, and Firmicutes are the predominant life forms in the studied springs. Archaea mainly include the phyla Euryarchaeota, Crenarchaeota, and Thaumarchaeota, and comprise less than 1% of the prokaryotic community. Comparison of microbial diversity in springs from Karvachar with that described for other terrestrial hot springs revealed that Proteobacteria, Bacteroidetes, Actinobacteria, and Deinococcus–Thermus are the common bacterial groups in terrestrial hot springs. Contemporaneously, specific bacterial and archaeal taxa were observed in different springs. Evaluation of the carbon, sulfur, and nitrogen metabolism in these hot spring communities has revealed diversity in terms of metabolic activity. Temperature seems to be an important factor in shaping the microbial communities of these springs. Overall, the diversity and richness of the microbiota are negatively affected by increasing temperature. Other abiotic factors, including pH, mineralization, and geological history, also impact the structure and function of the microbial community. More than 130 bacterial and archaeal strains (Bacillus, Geobacillus, Parageobacillus, Anoxybacillus, Paenibacillus, Brevibacillus Aeribacillus, Ureibacillus, Thermoactinomyces, Sporosarcina, Thermus, Rhodobacter, Thiospirillum, Thiocapsa, Rhodopseudomonas, Methylocaldum, Desulfomicrobium, Desulfovibrio, Treponema, Arcobacter, Nitropspira, and Methanoculleus) have been reported, some of which may be representative of novel species (sharing 91–97% sequence identity with their closest matches in GenBank) and producers of thermozymes and biomolecules with potential biotechnological applications. Whole-genome shotgun sequencing of T. scotoductus K1, as well as of the potentially new Treponema sp. J25 and Anoxybacillus sp. K1, were performed. Most of the phyla identified by 16S rRNA were also identified using metagenomic approaches. Detailed characterization of thermophilic isolates indicate the potential of the studied springs as a source of biotechnologically valuable microbes and biomolecules.

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