scholarly journals Screening and Characterization of Thermostable Amylase-Producing Bacteria Isolated from Soil Samples of Afdera, Afar Region, and Molecular Detection of Amylase-Coding Gene

2021 ◽  
Vol 2021 ◽  
pp. 1-14
Author(s):  
Semira Nureddin Yassin ◽  
Tamene Milkessa Jiru ◽  
Meera Indracanti
Keyword(s):  
Maximum Activity ◽  
Soil Samples ◽  
Effect Of Temperature ◽  
Bacterial Isolates ◽  
Original Activity ◽  
Zone Formation ◽  
Afar Region ◽  
Wide Range ◽  
Thermostable Amylase

Studying thermostable amylase-producing bacteria in extreme environments has a crucial role to overcome different industrial challenges. Afar Region is one of the hottest and salty areas, making it the home of extremophiles. This study aimed at screening and characterizing amylase-producing bacteria isolated from soil samples of Afdera, Afar Region, and detection of their amylase-coding genes. Thus, a total of 49 bacterial isolates were obtained from the collected soil samples. Out of these, three isolates (M2, M8, and M13) were selected on the basis of diameter of the average clear zone formation and time taken to decolorize iodine solution. Based on their morphological and biochemical characteristics, the isolates were identified as genus Bacillus. PCR amplification and detection of the amylase-coding gene confirmed the presence of the amylase gene in the three bacterial isolates. Optimum amylase production time for these isolates was 48 hrs (M13 and M8) and 72 hrs (M2) corresponding to the amylase activity of 0.67 U/mL for M13, 0.74 U/mL for M8, and 0.73 U/mL for M2 with an optimum temperature of 55°C. Studies on the effect of temperature revealed that the crude enzyme had a maximum activity and stability at 75°C, 70°C, and 65°C for isolates M13, M8, and M2, respectively. Additionally, amylase produced from all isolates retained more than 66.41% of their original activity after incubating them at a temperature range from 55 to 80°C for 50 min. Optimum pH for the activity of all crude amylases was in the range from 5 to 9 with a peak activity at pH 8. Their activity decreased significantly by the presence of Zn+2 and Mg2+; however, their activity increased by the presence of Ca+2. Moreover, the three crude amylases were stable (0–3 M) with NaCl concentration. Amylases of this finding with thermophilic and halophilic characteristics offer a wide range of applications in food, brewing, textile, starch, paper, and deterrent industries. Thus, identification of these Bacillus isolates at a molecular level and purification as well as detailed characterization of the types of amylases are recommended for effective utilization in different industries.

Purification and Characterization of Limit Dextrinase from Malted Barley

2012 ◽  
Vol 550-553 ◽  
pp. 1747-1754
Author(s):  
Ya Li Peng ◽  
Fei Hu
Keyword(s):  
Metal Ion ◽  
Ph Value ◽  
Maximum Activity ◽  
Effect Of Temperature ◽  
Purification And Characterization ◽  
Crude Extracts ◽  
Extraction And Purification ◽  
Sds Page ◽  
Limit Dextrinase

Limit dextrinase is one of three main amylases in malted barley, which plays a significant role during the mashing stage of brewing. Due to very low content and similar properties compared to other amylases in malted barley, limit dextrinase is hard to separate effectively. Our work had been directed towards the extraction and purification of limit dextrinase from malted barley. Final products were obtained through fraction precipitation with ammonium sulfate and column chromatography, and purified limit dextrinase acquired a high purity of 31.23 times as much as that of crude extracts. The previous results were also confirmed by sodiumdodecyl sulphate poly-acrylamide gel electrophoresis (SDS-PAGE) revealing a single band of protein (~97KDa). Effect of temperature, pH value, and metal ion on hydrolysis characterization of limit dextrinase was investigated. The results indicated that the maximum activity of purified samples changed significantly compared with that of crude extracts. The activity of purified limit dextrinase could be activated by lower concentration of Mg2+、Ca2+、Mn2+ and inhibited by the action of Zn2+、Fe2+. But this influence was not so obvious for K+.

scholarly journals Isolation and Characterization of Thermophilic Bacteria Indigenous to Al-Ahsa Desert

2020 ◽  
Vol 14 (3) ◽  
pp. 2157-2163
Author(s):  
Ahmed Anwar Al-Mulla ◽  
Ashraf Khalifa
Keyword(s):  
Standard Method ◽  
Thermophilic Bacteria ◽  
Soil Samples ◽  
Biotechnological Applications ◽  
Bacterial Isolates ◽  
Phenotypic Characteristics ◽  
Thermostable Enzymes ◽  
Biological Resources ◽  
Isolation And Characterization

Deciphering the biological resources across the Saudi niches is highly recommended for the prosperity. To this end, the aim of the current work was to isolate thermophilic bacteria from unexplored areas of Al-Ahsa region, and investigate their phenotypic characteristics. Three soil samples were collected from different desert sites of Al-Ahsa region. Thermophilic bacteria were isolated directly for soil samples into Thermus medium broth as a standard method. Single colonies of the actively growing bacterial isolates were preserved in 20% glycerol then kept at -80°C. The isolates were screened for production of thermostable enzymes using the commercially available kit API20E strip (bioMerieux, Marcy l’Etoile, France). Incubation were carried out at 50°C. It can be concluded that thermophilic bacteria in Al-Ahsa region harbor novel thermostable enzymes that might have biotechnological applications, in future.

ISOLATION, SCREENING AND CHARACTERIZATION OF AMYLASE AND CELLULASE PRODUCING BACTERIAL ISOLATES FROM DIFFERENT SOIL SAMPLES.

2017 ◽  
Vol 5 (10) ◽  
pp. 1385-1396
Author(s):  
Manthena Navabharath ◽  
◽  
Soumya a ◽  
Prajwala d ◽  
Venkatesh s ◽  
...  
Keyword(s):  
Soil Samples ◽  
Bacterial Isolates

Isolation and Characterization of Extracellular Enzyme (Glutaminase and Urease) producing Bacteria isolated from Soil Samples of Different Regions of Gwalior, Madhya Pradesh, India

2021 ◽  
Vol 16 (7) ◽  
pp. 122-129
Author(s):  
Neha Sharma ◽  
Shuchi Kaushik ◽  
Rajesh Singh Tomar
Keyword(s):  
Extracellular Enzyme ◽  
Soil Samples ◽  
Dilution Method ◽  
Bacterial Isolates ◽  
Alternative Source ◽  
Isolation And Characterization ◽  
Microbiological Laboratory ◽  
Biochemical Testing ◽  
Industrial Level

Microbial glutaminase and urease have demonstrated their benefits in various fields like medicinal, pharmaceutical and biotechnological industries. Keeping this viewpoint, the aim of the present study was the isolation and characterization of extracellular enzyme-producing bacteria from soil samples collected from different regions of Gwalior (M.P.). The isolated bacterial cultures were processed by serial dilution method and maintained on nutrient agar medium following standard microbiological laboratory practices for maintenance and preservation of bacteria. We screened out three enzyme producing strains of Salmonella sp., Proteus vulgaris and Bacillus subtilis. The screening was based on biochemical testing and enzyme assays. To accomplish this work, we used differential as well as selective media. All the selected isolates were able to produce enzymes like L-Glutaminase and Urease with different specific enzymatic activity. These bacterial isolates were not reported to show any type of allergenicity when their sequences were checked by bioinformatics tool Algpred. So, these bacterial isolates can be considered as an alternative source for the production of enzymes and can be used for largescale production of enzymes at the industrial level.

scholarly journals Characterization of Thermostable Amylase From Halophilic Lactobacillus Plantarum TS1

2021 ◽  
Author(s):  
Sania Riaz ◽  
Anum Fareed ◽  
Habiba Zaffar ◽  
Shafique Ur Rehman ◽  
Muhammad Jamil ◽  
...  
Keyword(s):  
Lactobacillus Plantarum ◽  
Saline Soil ◽  
Salt Range ◽  
Sds Page ◽  
Wide Range ◽  
Two Temperatures ◽  
Harsh Conditions ◽  
Thermostable Amylase ◽  
Important Enzyme

Abstract Amylase is an important enzyme use extensively in various industrial processes. It is mainly involved in the catalysis of starch that requires harsh conditions; therefore, it is required to isolate amylases with unique properties that makes it more applicable. Extremophiles are the major resource of such enzymes; therefore, amylase positive strains were isolated from the saline soil where the temperature is also exceptionally high. Five amylase positive strains were isolated from the Karak salt range, Kohat Pakistan and were identified by phylogenetic analysis. DNS based assay was employed to compare the activities of different amylases obtained from five strains while using the starch as a substrate. The amylase obtained from Lactobacillus plantarum TS1 was found to be more efficient, which was purified and characterized. The SDS-PAGE of purified amylase showed a single band with an estimated size of 10 kDa. The kinetic parameters were measured at two temperatures i.e. at 37 °C and 50 °C. The Kcat as well as the Kcat/KM were found to be high when temperature increased from 37 °C to 50 °C. Amylase was active at wide range of temperature as well as pH and work efficiently in the presence of salts and various organic solvents.

scholarly journals Isolation and characterization of a thermostable alkaline chitinase-producing Aeromonas strain and its potential in biodegradation of shrimp shell wastes

2021 ◽  
Vol 26 (2) ◽  
pp. 2511-2522
Author(s):  
ZHENGANG MA ◽  
◽  
JINFENG TONG ◽  
YAN WANG ◽  
ZEYANG ZHOU ◽  
...  
Keyword(s):  
Morphological Analysis ◽  
Maximum Activity ◽  
Precipitation Method ◽  
Chitinolytic Activity ◽  
Isolation And Characterization ◽  
Aeromonas Strain ◽  
Shrimp Shell ◽  
Wide Range ◽  
Culturing Conditions

Chitinases are employed to the conversion of chitin and are produced by a wide range of bacteria. The objective of this study was to isolate chitinase-producing microorganisms with high chitinolytic activity. A thermostable alkaline chitinase producing isolate strain CQNU6-2 was obtained from soil samples and showed potential in biodegradation of shrimp shell wastes. The optimal culturing conditions of isolate CQNU6-2 is at 25°C and pH 7 for 24 h. The chitinase produced by strain CQNU6-2 exhibited maximum activity at pH 6.0 and 40°C and it could tolerate the treatment of high temperature (up to 80°C) and high pH (over 10). Taxonomic study, based on biochemical and morphological analysis and phylogenetic analysis of 16S rDNA, showed that strain CQNU6-2 was belongs to the genus Aeromonas sp. The isolate can effectively hydrolyze colloidal chitin with degradation rate of 100% and also can directly degrade the shrimp shells. Ammonium sulfate precipitation method can be used to preliminary purify the chitinase. In conclusion, strain CQNU6-2 had a promising potential for biodegradation of chitin under harsh pH or temperature conditions and could be employed to the comprehensive utilization of shrimp shell wastes.

scholarly journals Screening of ?-amylase producing bacteria from tannery wastes of Hazaribag, Bangladesh

2017 ◽  
Vol 5 (2) ◽  
pp. 1-10
Author(s):  
Md Rayhan Islam ◽  
Omit Kumer Mondol ◽  
Md Saimoon Rahman ◽  
Md Morsaline Billah ◽  
Mohammad Shahedur Rahman ◽  
...  
Keyword(s):  
Amylase Activity ◽  
Biochemical Properties ◽  
Maximum Activity ◽  
Bacterial Isolates ◽  
Bacterial Strains ◽  
16S Rrna Sequence ◽  
16S Rrna Sequence Analysis ◽  
Tannery Wastes ◽  
Simple Sugar

Alpha amylases (?-amylases) are one of the most imperative enzymes for producing simple sugar units from complex sugar molecules. Attempts were made to isolate amylolytic bacterial strains from soil samples of tannery wastes collected from Hazaribagh, Dhaka and subsequent partial characterization was performed. Bacterial isolates were primarily screened for ?- amylase activity on starch agar medium. Based on microscopic and biochemical properties of isolates, ?-amylase activity of bacterial isolates were determined to find out two best producers of the enzyme. Subsequent molecular identification of these two ?-amylase producing bacterial isolates using 16s rRNA sequence analysis showed that isolates were Bacillus amyloliquefaciens and B. subtilis respectively. In submerged fermentation the B. amyloliquefaciens showed the highest activity (2.13 U/ml) while B. subtilis showed the second highest activity (1.89 U/ml). Characterization of the enzyme produced by B. amyloliquefaciens revealed that the maximum activity demonstrated at incubation time 25 min, pH 7.0 and temperature 500C. This newly isolated B. amyloliquefaciens could be exploited for the industrial production of ?-amylase with commercial implications.Jahangirnagar University J. Biol. Sci. 5(2): 1-10, 2016 (December)

scholarly journals Characterization of SdGA, a cold-adapted glucoamylase from Saccharophagus degradans

2021 ◽  
Author(s):  
Natael M. Wayllace ◽  
Nicolas Hedín ◽  
María V. Busi ◽  
Diego F. Gomez-Casati
Keyword(s):  
Marine Bacterium ◽  
Catalytic Domain ◽  
Biochemical Properties ◽  
Maximum Activity ◽  
High Rate ◽  
Saccharophagus Degradans ◽  
Cold Adapted ◽  
Structural And Functional Properties ◽  
Wide Range

ABSTRACTWe investigated the structural and functional properties of SdGA, a glucoamylase (GA) from Saccharophagus degradans, a marine bacterium which degrades different complex polysaccharides at high rate. SdGA is composed mainly by a N-terminal GH15_N domain linked to a C-terminal catalytic domain (CD) found in the GH15 family of glycosylhydrolases with an overall structure similar to other bacterial GAs. The protein was expressed in Escherichia coli cells, purified and its biochemical properties were investigated. Although SdGA has a maximum activity at 39°C and pH 6.0, it also shows high activity in a wide range, from low to mild temperatures, like cold-adapted enzymes. Furthermore, SdGA has a higher content of flexible residues and a larger CD due to various amino acid insertions compared to other thermostable GAs. We propose that this novel SdGA, is a cold-adapted enzyme that might be suitable for use in different industrial processes that require enzymes which act at low or medium temperatures.

scholarly journals Screening and characterization of Phosphorus solubilizing Bacteria and their effect on Rice seedlings

2015 ◽  
Vol 1 (1) ◽  
pp. 27-35 ◽  
Author(s):  
Md Shahinur Rahman ◽  
Quazi Forhad Quadir ◽  
Atiqur Rahman ◽  
Moonmoon Nahar Asha ◽  
Md Abul Khair Chowdhury
Keyword(s):  
Soil Samples ◽  
Bacterial Isolates ◽  
Rice Seedlings ◽  
Gram Negative ◽  
P Content ◽  
Pure Cultures ◽  
Plant Trial ◽  
Solubilizing Capacity

An experiment was carried out to isolate, screen and characterize bacteria collected from an industrially polluted site of Bhaluka under the Mymensingh district and to evaluate their phosphorus (P) solubilizing capacity. About ten plant and soil samples from six different spots were collected from the site. Thirty four bacterial isolates were screened and pure cultures of the different bacterial isolates were prepared. Among the bacterial isolates 25 were gram negative and 9 were gram positive. About 31 bacterial isolates had catalase producing capacity and remaining 3 were negative to catalase test. Bacterial isolates were grown on a NBRIP media to determine their phosphorus solubilizing capacity. About 25 bacterial isolates were shown P solubilizing capacity. Isolate SB8 gave the highest result about 11.42 PSI (phosphorus solubilizing index), whereas other bacterial isolates showed moderate P solubilizing capacity (PSI 1.75-6.35). A plant trial with selected isolates (SB8, SB15, SB25) were also done and SB8 achieved 10% higher P content in comparison with control which supports the in vitro P solubilization assays. DOI: http://dx.doi.org/10.3329/ralf.v1i1.22353 Res. Agric., Livest. Fish.1(1): 27-35, Dec 2014

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