Optimization, Production, and Partial Purification of Thermostable -Amylase Produced by Marine Bacterium Bacillus sp. NRC12017

2017 ◽  
Vol 9 (08) ◽  
Author(s):  
Ebtsam M. El-Kady ◽  
Mohsen S. Asker ◽  
Saadia M. Hassanein ◽  
Eman A. Elmansy ◽  
Fawkia M. El-Beih
Keyword(s):  
Yeast Extract ◽  
Fermentation Broth ◽  
Maximum Yield ◽  
Thermophilic Bacterium ◽  
Inoculum Size ◽  
Ph Stability ◽  
Partial Purification ◽  
Bacillus Sp ◽  
Main Band ◽  
Moderately Thermophilic

α-Amylase biosynthesis was illustrated under stress circumstances of high temperature and high salinity in aerobically cultivated culture of a newly isolated moderately thermophilic bacterium of spore-forming Bacillus sp. NRC12017 in medium containing starch, peptone and yeast extract. Maximum yield (18.41 U/ml) was took place at pH 6.5 with 200 µl inoculum size at 45°C and an incubation time of 3 days. The ideal volume of the fermentation broth was found to be 15 ml in 100 ml Erlenmeyer flask, supplementation of starch at 2.5% and peptone plus yeast extract at 0.7% brought about the greatest production of α-amylase. Two fractions of α-amylase activities, designated FIII and FIV, were refined from culture filtrate utilizing ammonium sulfate (60 and 80%) and showed the main band at 30 KDa. Both fractions had the highest activity at 250 µl starch in a reaction mixture and pH 6.0-7.0. FIII and FIV showed that a temperature of 50 and 55°C and a reaction time of 20 and 30 min were the best available conditions for their activities, respectively, both fractions were stable up to 65°C and the activity was decreased drastically to 3.22 and 3.26 % when heated at 70°C. Concerning pH stability, a broad range of pH stability (5.0-11.0) was obtained by FIII and FIV.

scholarly journals First Insights into the Genome of the Moderately Thermophilic Bacterium Clostridium tepidiprofundi SG 508 T

2016 ◽  
Vol 4 (3) ◽  
Author(s):  
Anja Poehlein ◽  
Ines Friedrich ◽  
Larissa Krüger ◽  
Rolf Daniel
Keyword(s):  
Yeast Extract ◽  
Hydrothermal Vent ◽  
Thermophilic Bacterium ◽  
Gram Positive ◽  
Moderately Thermophilic ◽  
Beef Extract ◽  
Clostridial Cluster

The moderately thermophilic bacterium Clostridium tepidiprofundi is Gram-positive and belongs to clostridial cluster I. It was isolated from a hydrothermal vent chimney. Substrates utilized by C. tepidiprofundi include casein, peptone, tryptone, yeast extract, beef extract, starch, maltose, and glucose. The genome consists of one replicon (3.06 Mb).

scholarly journals Phenylobacterium lituiforme sp. nov., a moderately thermophilic bacterium from a subsurface aquifer, and emended description of the genus Phenylobacterium

2004 ◽  
Vol 54 (6) ◽  
pp. 2141-2146 ◽  
Author(s):  
Sungwan Kanso ◽  
Bharat K. C. Patel
Keyword(s):  
Yeast Extract ◽  
Optimal Growth ◽  
Thermophilic Bacterium ◽  
Wall Structure ◽  
Rrna Gene ◽  
Thin Sections ◽  
Gram Negative ◽  
Moderately Thermophilic ◽  
Registration Number ◽  
Flowing Waters

A facultative anaerobic bacterium, strain FaiI3T, was isolated from samples collected from the free-flowing waters of a bore well (Fairlea Bore, registration number 3768) which taps into the Australian Great Artesian Basin subsurface thermal aquifer. Strain FaiI3T developed yellow to pale-yellow colonies (0·5–1·5 mm) after 48 h. The non-spore forming rods (0·5×1–3 μm) were slightly curved, occurred singly and as pairs and were motile with a single polar flagellum. Cells tended to form clumps in liquid medium and rosettes were commonly observed. The cells stained Gram-negative and electron micrographs of thin sections revealed a multi-layered complex Gram-negative cell wall structure. Strain FaiI3T grew optimally at 40–41 °C, with growth observed at 45 °C but not at 50 °C. The pH growth range was between pH 6 and 9 and optimal growth occurred between pH 6 and 6·5. Strain FaiI3T grew best with yeast extract as the sole carbon and energy source. Peptone, yeast extract, acetate, xylose, sucrose, glucose, glycerol, succinate, butyrate, lactate, fumarate, citrate, l-phenylalanine, cellobiose and gelatin supported growth but maltose, fructose, glycine, ethanol, benzoate and oxalate did not. Tyrosine was produced from l-phenylalanine. Strain FaiI3T was catalase-positive and oxidase-negative and did not hydrolyse starch. Growth was inhibited by neomycin, tetracycline, streptomycin, chloramphenicol, ampicillin, vancomycin and spectinomycin. The G+C content was determined to be 66·5±0·5 mol%. On the basis of the 16S rRNA gene sequence analysis, strain FaiI3T was assigned as a novel species of the genus Phenylobacterium, Phenylobacterium lituiforme sp. nov. in the order Caulobacterales, subclass α-Proteobacteria, class Proteobacteria. The type strain is FaiI3T (=ATCC BAA-294T=DSM 14363T).

scholarly journals Effect of the medium composition on formation of amylase by Bacillus sp

2003 ◽  
Vol 46 (1) ◽  
pp. 129-134 ◽  
Author(s):  
Eliana de Oliveira. Santos ◽  
Meire Lelis Leal Martins
Keyword(s):  
Carbon Source ◽  
Yeast Extract ◽  
Soluble Starch ◽  
Medium Composition ◽  
Alpha Amylase ◽  
Bacillus Sp ◽  
Glucose Effect ◽  
Moderately Thermophilic ◽  
Exogenous Glucose ◽  
Culture Supernatants

Studies on the alpha -amylase synthesis was carried out with a moderately thermophilic, facultatively anaerobic Bacillus sp, isolated from soil samples. The cells were cultivated in a complex medium containing soluble starch or maltose as carbon source. The levels of the alpha -amylaseactivity detected in culture supernatants varied greatly with the type of carbon source used. Maltose, soluble starch and citrate stimulated alpha -amylaseformation. Addition of exogenous glucose repressed formation of alpha -amylase, demonstrating that a classical glucose effect was operative in this organism. The concentration of yeast extract was found to be important factor in the alpha -amylase synthesis bythe isolate.The activity of the enzyme increased between 2 and 5 g/L yeast extract concentration and then fell very rapidly beyond this point. The best concentration of peptone to alpha-amylase formation was found to be around 10g/L.

scholarly journals Screening and Selection of Media Components for Protease Production by Bacillus sp. EBTA6 Using Plackett–Burman Design

2020 ◽  
Vol 8 (6) ◽  
pp. 1250-1255
Author(s):  
Fikriye Alev Akçay ◽  
Ayşe Avcı
Keyword(s):  
Yeast Extract ◽  
Protease Activity ◽  
Inoculum Size ◽  
Protease Production ◽  
Bacillus Sp ◽  
Medium Components ◽  
Dipotassium Phosphate ◽  
Burman Design ◽  
Plackett Burman Design ◽  
Selection Of

In this study, effects of medium components and inoculum size on the protease production by Bacillus sp. EBTA6 that was isolated from a home-made Tarhana sample were investigated. The cell-free supernatant of bacterium cultured on a shaking incubator for 24 h was used to test protease activity as the response. With a total number of 11 factors, 12 different experiments were run and the highest experimental protease activity was measured as 2280.4 U/mL. Results were analyzed statistically by ANOVA and the most efficient factors were detected as yeast extract, dipotassium phosphate, casein, and peptone with a contribution of 93.78, 2.19, 1.96, 1.31%, respectively. For validation of the selected factors, a further experiment was performed by using of yeast extract (9.98 g/L), dipotassium phosphate (1.27 g/L), casein (8.69 g/L), and peptone (9.88 g/L) obtained from the design equation. The experimental response was found as 2411.4 U/mL which was only 5.5% higher than the predicted response showing that the model was applicable.

Convenient Partial Purification of Glucose Oxidase from Aspergillus niger A9 Fermentation Broth by Reversed Micelles

2012 ◽  
Vol 554-556 ◽  
pp. 957-961
Author(s):  
Hong An ◽  
Xi Feng He ◽  
Shu Gang Gao
Keyword(s):  
Enzyme Activity ◽  
Aspergillus Niger ◽  
Glucose Oxidase ◽  
Fermentation Broth ◽  
Volume Ratio ◽  
Reversed Micelles ◽  
Partial Purification ◽  
Ammonium Bromide ◽  
Ultrasonic Oscillation ◽  
Protein Activity

Aim of this work was to establish the optimum conditions for the extraction and recovery by cationic reversed micelles of glucose oxidase (GOX) from Aspergillus niger A9, The influence of pH, temperature, solvent/co-solvents ratio on the extraction was investigated by experiment, using the residual enzyme activity to evaluate the results. The best condition for GOX extraction were ensured using iso-octane as solvent and butanol and n-hexanol co-solvent at 76/18/6 volume ratio, pH 4.80, 200mM cetyl-trimethyl ammonium bromide (CTAB) as cationic surfactant, The enzyme activity of GOX is measured by DNS method (3,5-dinitro salicylic acid method). In the extraction process, ultrasonic oscillation was adopted to mix organic solvent and water, ultrasonic oscillation temperature is 45 °C. Protein activity recovery of GOX can reach 88.2% in extraction.

Identification and Characterization of a New Alkaline SGNH Hydrolase from a Thermophilic Bacterium Bacillus sp. K91

2016 ◽  
Vol 26 (4) ◽  
pp. 730-738 ◽  
Author(s):  
Tingting Yu ◽  
Junmei Ding ◽  
Qingxia Zheng ◽  
Nanyu Han ◽  
Jialin Yu ◽  
...  
Keyword(s):  
Thermophilic Bacterium ◽  
Bacillus Sp ◽  
Bacterium Bacillus ◽  
Identification And Characterization

scholarly journals Response Surface Methodology for the Optimization of Keratinase Production in Culture Medium Containing Feathers by Bacillus sp. UPM-AAG1

Catalysts ◽  
2020 ◽  
Vol 10 (8) ◽  
pp. 848 ◽  
Author(s):  
Aa’ishah Abdul Gafar ◽  
Mohd Ezuan Khayat ◽  
Siti Aqlima Ahmad ◽  
Nur Adeela Yasid ◽  
Mohd Yunus Shukor
Keyword(s):  
Amino Acids ◽  
Proteolytic Enzyme ◽  
Culture Medium ◽  
Essential Amino Acids ◽  
Inoculum Size ◽  
Commercial Application ◽  
Bacillus Sp ◽  
Broad Application ◽  
Feather Waste ◽  
Central Composite

Keratinase is a type of proteolytic enzyme with broad application in industry. The main objective of this work is the optimization of keratinase production from Bacillus sp. strain UPM-AAG1 using Plackett-Burman (PB) and central composite design (CCD) for parameters, such as pH, temperature, feather concentration, and inoculum size. The optimum points for temperature, pH, and inoculum and feather concentrations were 31.66 °C, 6.87, 5.01 (w/v), and 4.53 (w/v), respectively, with an optimum keratinase activity of 60.55 U/mL. The keratinase activity was further numerically optimized for commercial application. The best numerical solution recommended a pH of 5.84, temperature of 25 °C, inoculums’ size of 5.0 (v/v), feather concentration of 4.97 (w/v). Optimization resulted an activity of 56.218 U/mL with the desirability value of 0.968. Amino acid analysis profile revealed the presence of essential and non-essential amino acids. These properties make Bacillus sp. UPM-AAG1 a potential bacterium to be used locally for the production of keratinase from feather waste.

scholarly journals Production of Itaconic Acid from Jatropha curcas Seed Cake by Aspergillus terreus

2013 ◽  
Vol 5 (1) ◽  
pp. 57-61 ◽  
Author(s):  
Amina M. AHMED EL-IMAM ◽  
Muinat O. KAZEEM ◽  
Mutiat B. ODEBISI ◽  
Mushaffa A. OKE ◽  
Azeezat O. ABIDOYE
Keyword(s):  
Itaconic Acid ◽  
Jatropha Curcas ◽  
Substrate Concentration ◽  
Aspergillus Terreus ◽  
Maximum Yield ◽  
Inoculum Size ◽  
Daily Maximum ◽  
Dilute Acid Hydrolysis ◽  
Seed Cake ◽  
Jatropha Seed

Submerged substrate fermentation of Jatropha seed cake, a by-product of oil extraction from Jatropha curcas seed was carried out using Aspergillus terreus for the production of itaconic acid. The Jatropha seed cake was initially converted into fermentable sugars by dilute acid hydrolysis using 50% sulphuric acid. The rate of hydrolysis was 1.04 gL-1. The fermentation process was carried out at room temperature, agitation of 400 rpm and three physico-chemical parameters (pH, inoculum size and substrate concentration) were varied. Itaconic acid and glucose assays were carried out by spectrophotometry and Dinitrosalicylic acid methods respectively daily. Maximum yield of itaconic acid was 48.70 gL-1 at 5 ml of inoculum size, 50 g substrate concentration and pH 1.5. The residual glucose concentration increased for the first two days of fermentation after which it began to decrease as the itaconic acid concentration increased. The least concentration of itaconic acid observed was 6.00 gL-1, obtained after 24 hours of fermentation with 4 ml inoculum size, 50 g substrate concentration and at pH 1.5. The findings of this work indicate that Jatropha curcas seed cake is a suitable substrate for itaconic acid production.

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