scholarly journals Isolation and Characterisation of Thermophilic Bacillus licheniformis SUNGC2 as Producer of α-Amylase from Malaysian Hot Spring

2020 ◽  
Vol 28 (S2) ◽  
Author(s):  
Marwan Jawad Msarah ◽  
Ayesha Firdose ◽  
Izyanti Ibrahim ◽  
Wan Syaidatul Aqma
Keyword(s):  
Bacillus Licheniformis ◽  
Specific Activity ◽  
Hot Spring ◽  
Ammonium Phosphate ◽  
Fold Increase ◽  
Relative Activity ◽  
Biochemical Properties ◽  
Culture Conditions ◽  
Thermophilic Microorganisms ◽  
Amylase Production

Screening of new source of novel and industrially useful enzymes is a key research pursuit in enzyme biotechnology. The study aims to report the characteristics of novel thermophilic microorganisms isolated from Sungai Klah (SK) Hot Spring, Perak, Malaysia, that can produce α-amylase. The morphological and biochemical properties were examined for SUNGC2 sample. The isolate was further screened for amylase, followed by 16S rRNA and analytical profile index (API) test. This isolate was further subjected to pH optimisation for α-amylase production. It was found that SUNGC2 was an α-amylase producer and was identified as Bacillus licheniformis SUNGC2 with NCBI accession numbers MH062901. The enzyme was found to exhibit an optimum temperature of 50°C and a pH of 7.0. The relative activity of the enzyme was obtained based on the improvement of the culture conditions. The highest amount of amylase production was 24.65 U/mL at pH 7.0, consecutively the growth was also highest at pH 7.0 with a 9.45-fold increase in specific activity by ammonium phosphate precipitation of 80% (w/v). The results showed that the bacteria isolated from the hot spring are a significant source of thermophilic enzymes that are highly promising in biotechnology.

scholarly journals Studies on the production of glucose isomerase by Bacillus licheniformis

2015 ◽  
Vol 17 (3) ◽  
pp. 84-88 ◽  
Author(s):  
Ogbonnaya Nwokoro
Keyword(s):  
Enzyme Activity ◽  
Bacillus Licheniformis ◽  
Organic Nitrogen ◽  
Specific Activity ◽  
Inorganic Nitrogen ◽  
Glucose Isomerase ◽  
Culture Conditions ◽  
Enzyme Productivity ◽  
Carbon Substrates ◽  
Optimum Ph

Abstract This work reports the effects of some culture conditions on the production of glucose isomerase by Bacillus licheniformis. The bacterium was selected based on the release of 3.62 mg/mL fructose from the fermentation of glucose. Enzyme was produced using a variety of carbon substrates but the highest enzyme activity was detected in a medium containing 0.5% xylose and 1% glycerol (specific activity = 6.88 U/mg protein). Media containing only xylose or glucose gave lower enzyme productivies (specific activities= 4.60 and 2.35 U/mg protein respectively). The effects of nitrogen substrates on glucose isomerase production showed that yeast extract supported maximum enzyme activity (specific activity = 5.24 U/mg protein). Lowest enzyme activity was observed with sodium trioxonitrate (specific activity = 2.44 U/mg protein). In general, organic nitrogen substrates supported higher enzyme productivity than inorganic nitrogen substrates. Best enzyme activity was observed in the presence of Mg2+ (specific activity = 6.85 U/mg protein) while Hg2+ was inhibitory (specific activity = 1.02 U/mg protein). The optimum pH for best enzyme activity was 6.0 while optimum temperature for enzyme production was 50ºC.

scholarly journals Chloride Activated Halophilic α-Amylase from Marinobacter sp. EMB8: Production Optimization and Nanoimmobilization for Efficient Starch Hydrolysis

2015 ◽  
Vol 2015 ◽  
pp. 1-9 ◽  
Author(s):  
Sumit Kumar ◽  
S. K. Khare
Keyword(s):  
Immobilized Enzyme ◽  
Fold Increase ◽  
Culture Conditions ◽  
Production Optimization ◽  
Cross Linking ◽  
Functionalized Silica ◽  
Potential Source ◽  
Amylase Production ◽  
Repeated Use ◽  
Functionalized Silica Nanoparticles

Halophiles have been perceived as potential source of novel enzymes in recent years. The interest emanates from their ability to catalyze efficiently under high salt and organic solvents. Present work encompasses production optimization and nanoimmobilization of an α-amylase from moderately halophilic Marinobacter sp. EMB8. Media ingredients and culture conditions were optimized by “one-at-a-time approach.” Starch was found to be the best carbon source at 5% (w/v) concentration. Glucose acted as catabolic repressor for amylase production. Salt proved critical for amylase production and maximum production was attained at 5% (w/v) NaCl. Optimization of various culture parameters resulted in 48.0 IU/mL amylase production, a 12-fold increase over that of unoptimized condition (4.0 IU/mL). α-Amylase was immobilized on 3-aminopropyl functionalized silica nanoparticles using glutaraldehyde as cross-linking agent. Optimization of various parameters resulted in 96% immobilization efficiency. Starch hydrolyzing efficiency of immobilized enzyme was comparatively better. Immobilized α-amylase retained 75% of its activity after 5th cycle of repeated use.

scholarly journals Production optimization using Plackett-Burman and Box-Behnken designs with partial characterization of amylase from marine actinomycetes

2021 ◽  
Author(s):  
Sarah I Bukhari ◽  
Mohamed H Al-Agamy ◽  
Mahmoud S Kelany ◽  
Mohammad R Al Hazani ◽  
Moaz M Hamed
Keyword(s):  
Molecular Weight ◽  
Specific Activity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Fold Increase ◽  
Fermentation Medium ◽  
Culture Conditions ◽  
Production Optimization ◽  
Activity Measurement

Abstract Amylase is an industrial enzyme that is used in the food and biofuel industries. We screened four actinomycetes strains for amylase biosynthesis. The Streptomyces rochei strain had a larger hydrolytic zone (24 mm) on starch agar plates, than the other isolates. Plackett-Burman’s experimental design was implemented to optimize the conditions for amylase production by the selected strains. Growth under optimized culture conditions led to 1.7, 9.8, 7.7, and 3.12 -fold increases for the isolates S. griseorubens, S. rochei, S. parvus, and Streptomyces sp., respectively, in the specific activity measurement in comparison with growth under primary conditions. When applying the Box-Behnken design on S. rochei using the most significant parameters starch, K2HPO4, pH, and temperature, there was a 12.22-fold increase in the specific activity measurement: 7.37 U/mg. The optimal fermentation medium formula was kept at 30.6°C for seven days. The amylase from S. rochei was partially purified, and its molecular weight was determined using Sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The molecular weight was found to be 45, 43, and 53 kDa. Amylase was particularly active at pH 6 and 65°C. The purified enzyme was most active at 65°C and a pH of 6, thermal stability of 70°C for 40 min and salt concentration of 1 M with a Km and Vmax of 6.58 mg/ml and 21.93 mg/ml/min, respectively. The amylase improved by adding Cu + 2, Zn + 2, and Fe + 2 (152.21%, 207.24%, and 111.89%). Increased production of amylase enzyme by Streptomyces rochei KR108310 attracts the production of industrially significant products.

scholarly journals Production of thermostable extracellular α-amylase by a moderate thermophilic Bacillus licheniformis isolated from Qinarje Hot Spring (Ardebil prov. of Iran)

2017 ◽  
Vol 118 (4) ◽  
Author(s):  
Ali Deljou ◽  
Iman Arezi
Keyword(s):  
Carbon Source ◽  
Nitrogen Source ◽  
Bacillus Licheniformis ◽  
Bacterial Growth ◽  
Enzyme Production ◽  
Hot Spring ◽  
Basal Medium ◽  
Thermophilic Bacterium ◽  
Inoculum Size ◽  
Amylase Production

Background and Purpose: Amylases are most important industrial enzymes that account for about 30% of the world’s food, feed, fermentation, textile, detergent and cellulosic industries. This study aimed at optimum production of thermostable α-amylase via moderate thermophilic bacterium (Bacillus licheniformis) which was recently isolated from Qinarje Hot spring.Material and Methods: Initially, ability of bacterium for amylase activity was determined by starch hydrolysis test using Gram’s iodine staining. Then bacterial growth pattern and amylase production curves in basal production medium were graphically determined at different time intervals. Finally, effect of different temperature, pH, carbon source, nitrogen source, minerals and inoculum size were studied on bacterial growth and amylase production using turbidimetric and DNS method, respectively.Results: Optimum enzyme production achieved after 84 hours of inoculation from cultures growing at 40 ˚C and pH 9.0 in a medium containing 0.03% (w/v) of CaCl2, compared to the basal medium, results showed that the best enzyme production happened with inoculum size of 4% (v/v). The addition of 1% (w/v) rice husk (as a Carbon source) enhanced enzyme productivity up to 160% and substitution of the peptone and yeast extract with 1% (w/v) of tryptone (as a Nitrogen source) increased the α-amylase production up to 160%.Conclusion: Our findings show that B. licheniformis-AZ2 strain has an ability to produce the thermostable α-amylase which is suitable in starch processing and food industries. To be commercialized, further investigation is required for enhancement of the enzyme production.Keywords: Bacillus licheniformis; Optimization; Basal medium; Agricultural by-products.

scholarly journals Production and Partial Characterization of α-Amylase Enzyme from Marine Actinomycetes

2021 ◽  
Vol 2021 ◽  
pp. 1-15
Author(s):  
Mohamed H. Al-Agamy ◽  
Mohammad R. Alhuzani ◽  
Mahmoud S. Kelany ◽  
Moaz M. Hamed
Keyword(s):  
Specific Activity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Fold Increase ◽  
Culture Conditions ◽  
Activity Measurement ◽  
Terrestrial Environment ◽  
Box Behnken Design ◽  
Thermal Stability Of

Amylase producing actinobacteria were isolated and characterized from terrestrial environment. There are a limited number of reports investigating the marine environment; hence, in the present study, four marine enzymes were tested for their amylase production ability. On starch agar plates, the Streptomyces rochei strain showed a higher hydrolytic zone (24 mm) than the other isolates. Growth under optimized culture conditions using Plackett-Burman’s experimental design led to a 1.7, 9.8, 7.7, and 3.12-fold increase for the isolates S. griseorubens, S. rochei, S. parvus, and Streptomyces sp., respectively, in the specific activity measurement. When applying the Box-Behnken design on S. rochei using the most significant parameters (starch, K2HPO4, pH, and temperature), there was a 12.22-fold increase in the specific activity measurement 7.37 U/mg. The α-amylase was partially purified, and its molecular weight was determined using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. α-Amylase was particularly active at pH 6 and 65°C. The purified enzyme was most active at 65°C and pH 6, thermal stability of 70°C for 40 min, and salt concentration of 1 M with Km and Vmax of 6.58 mg/ml and 21.93 μmol/ml/min, respectively. The α-amylase was improved by adding Cu+2, Zn+2, and Fe+2 (152.21%, 207.24%, and 111.89%). Increased production of α-amylase enzyme by S. rochei KR108310 leads to production of significant industrial products.

scholarly journals Isolation, Biochemical Characterisation and Identification of Thermotolerant and Cellulolytic Paenibacillus lactis and Bacillus licheniformis

2021 ◽  
Vol 59 (3) ◽  
Author(s):  
Krzysztof Makowski ◽  
Martyna Leszczewicz ◽  
Natalia Broncel ◽  
Lidia Lipińska-Zubrycka ◽  
Adrian Głębski ◽  
...  
Keyword(s):  
Bacillus Licheniformis ◽  
Hot Spring ◽  
Growth Curves ◽  
Maximum Growth ◽  
Cellulolytic Activity ◽  
Bacterial Strains ◽  
Thermophilic Microorganisms ◽  
Biochemical Characteristics ◽  
Cmcase Activity ◽  
Biochemical Characterisation

Research background. Cellulose is an ingredient of waste materials that can be converted to other valuable substances. This is possible provided that, the polymer molecule will be degraded to smaller particles, used as a carbon source by microorganisms. Because of the frequently applied methods of pre-treatment of lignocellulosic materials, the cellulases derived from thermophilic microorganisms are particularly desirable. Experimental approach. We were looking for cellulolytic microorganisms able to grow at 50 °C. We described their morphological features and biochemical characteristics based on CMCase activity and the api®ZYM. The growth curves, during incubation at 50 °C, were examined using the microbioreactor BioLector®. Results and conclusions. 40 bacterial strains were isolated from fermenting hay, geothermal karst spring, hot spring and geothermal pond at 50 °C. The vast majority of the bacteria were Gram-positive and rod-shaped with the maximum growth temperature of at least 50 °C. We also demonstrated a large diversity of biochemical characteristics among the microorganism. The CMCase activity was confirmed for 27 strains. However, the hydrolysis capacities (HC) were significant in bacterial strains: BBLN1, BSO6, BSO10, BSO13 and BSO14, and reached 2.74, 1.62, 1.30, 1.38 and 8.02 respectively. Rapid and stable growth was presented, among others, by BBLN1, BSO10, BSO13 and BSO14. The strains fulfilled the selection conditions and were identified based on the 16S rDNA sequences. BBLN1, BSO10, BSO13 were classified as Bacillus licheniformis, whereas BSO14 as Paenibacillus lactis. Novelty and scientific contribution. We described cellulolytic activity and biochemical characteristics of many bacteria isolated from hot environments. We are also the first to report the cellulolytic activity of thermotolerant P.s lactis. Described strains can be a source of new thermostable cellulases, which are extremely desirable in various branches of the circular bioeconomy.

Further Studies On The Purified Phospholipase A2 From Human Platelet Plasma Membranes

1981 ◽  
Author(s):  
R Apitz-Castro ◽  
M R Cruz ◽  
M Mas ◽  
M K Jain
Keyword(s):  
Arachidonic Acid ◽  
Plasma Membrane ◽  
Phosphatidic Acid ◽  
Phospholipase A2 ◽  
Plasma Membranes ◽  
Specific Activity ◽  
Blood Platelets ◽  
Fold Increase ◽  
Biochemical Properties ◽  
Mole Percent

The activation of a phospholipase A2 has been postulated in the release of arachidonic acid from phospholipids of platelet plasma membrane. The present communication deals with the biochemical properties of a phospholipase A2 isolated from human blood platelets. Phospholipase A2 from human platelets have been isolated to homogeneity through an acid or salt extraction, followed by affinity chromatography purification. The enzyme is copurified with the platelet plasma membrane, It has a MW of about 50,000 Dalton. It shows an optimum pH at 9.4 and requires calcium for its activity. Although it attacks a variety of phospholipid substrates, there is a preference for unsaturated phospholipids. Diarachidonoy1-PC liposomes can be hydrolyzed in the absence of any additive. The specific activity is markedly affected by the quality of the interface, showing variations of more than 10 fold between different substrate forms. The purified enzyme is considerably more active on the aggregated form of the substrate than on the monomeric form. Saturation behavior, consistent with Michaelis-Menten type of kinetics is observed at higher concentration of the micellar substrate. A four-fold increase in rate is observed when the enzyme is assayed in the presence of fatty acid + lysophospholipid. Maximal rates are obtained at a 20 mole percent of products to substrate. 1,2-diglyceride and phosphatidic acid stimulates the hydrolysis of phosphatidylcholine by the purified enzyme, however, in these forms of substrate, neither the diglyceride nor phosphatidic acid is hydrolyzed. Under optimal condition, the phospholipase A2 activity corresponds to at least 13 nmol/min/109 platelets. Activation of this enzyme by some intermediate related to the phospholipase C pathway might play a role in the stimulus- linked release of platelet arachidonic acid.

XYLANASE, AN ENVIRONMENTALLY FRIENDLY BLEACHING AGENT FOR PULP AND PAPER: Characterization And Stabilization Study Of Bacillus licheniformis I-5 CRUDE Xylanase

2018 ◽  
Vol 7 (3) ◽  
Author(s):  
Budiasih Wahyuntari., dkk
Keyword(s):  
Thermal Stability ◽  
Bacillus Licheniformis ◽  
Hot Spring ◽  
Sodium Dodecyl ◽  
Luria Broth ◽  
Crude Enzyme ◽  
Half Time ◽  
Triton X100 ◽  
Sds Page ◽  
Xylanolytic Enzymes

Isolate I-5 was isolated from Ciseeng hot spring, West Java and was identified as Bacillus licheniformis I-5. The isolate produces extracellular xylanolytic enzymes on Oatspelt containing Luria broth agar medium. Optimal activity of the crude enzyme was  observed at 50ºC and pH 7. The effect of sodium dodecyl sulphate, b-mercaptoethanol and Triton-X100 were observed. Incubating the crude enzyme in 1.5% SDS and 1.5% b-mercaptoethanol at 50oC for 90 minutes then adding Triton-X100 at final concentration of 3.5% for 45 minutes only reduced 5.75% of the initial enzyme activity. SDS/PAGE and zymogram analysis showed that at least two xylanolytic enzymes presence in the crude enzyme. The molecular weight of the enzyme was estimated about 127 and 20kD. The enzyme hydrolysed xylan into xylobiose, xylotriose and other longer xylooligosaccharides. Thermal stability of the crude enzyme was observed at 50, 60, and 70oC and pH 7 and 8. The results showed that the half time of the crude enzyme incubated at 50, 60, and 70oC pH 7 was 2 hours 55 minutes; 2 hours 33 minutes and 1 hour 15 minutes respectively. The half time at 50, 60 and 70oC, pH 8 was 2 hours 48 minutes; 1 hour 22 minutes and 1 hour 9 minutes respectively.keywords: Xilanase, Bacillus licheniformis I-5, thermal stability

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