Comparative Characterization and Amylase Activity Assessment of Certain Garden Bacterial Isolates

This study aimed to isolate and identify bacteria that can produce amylase enzyme from the unexplored Nasinuan Forest, Kantarawichai District, Mahasarakham Province, Thailand. Thirteen bacterial isolates with amylase-producing capacity on 1% starch agar were identified using 16S rRNA sequencing. Twelve bacteria were gram-positive, rod shaped and identified as Bacillus spp. and one bacterium with gram-negative and rod shaped character was Enterobacter cloacae. Their closest relatives were found in India, China, Korea, Indonesia, Argentina, Italy, Israel, USA, Argentina and South Africa. These bacteria were tested for specific amylase activity after 1-3 days enzyme induction with 1% starch at 37°C. The results showed the highest specific activity at day 2 incubation in the order: Bacillus cereus 3.5AL2 > 3.4AL1 > 1.4AL3 and thus 2-day enzyme induction was chosen for further analysis. Bacillus sp. 3.5AL2 was found to exhibit the highest specific amylase enzyme activity of 1.97 ± 0.41 U/mg protein at the optimal conditions of 60°C and pH 7.0 after 30 min incubation with 1% starch in 0.05 M PBS buffer. This amylase–producing bacterial strain offers great potential for applications in food and agricultural industries in Thailand.

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The potential of amylase enzyme activity against bacteria isolated from several lakes in East Java, Indonesia

Biodiversitas Journal of Biological Diversity ◽

10.13057/biodiv/d220106 ◽

2020 ◽

Vol 22 (1) ◽

Author(s):

INDAH KHOIRUN NISA ◽

Prabaningtyas Sitoresmi ◽

BETTY LUKIATI ◽

RINA TRITURANI SAPTAWATI ◽

ACHMAD RODIANSYAH

Keyword(s):

Enzyme Activity ◽

Molecular Identification ◽

Bacterial Isolate ◽

Amylase Activity ◽

Sequence Similarity ◽

Rrna Gene ◽

Neighbor Joining ◽

Bacterial Isolates ◽

Bacillus Cereus Group ◽

The 16S Rrna Gene

Abstract. Nisa IK, Prabaningtyas S, Lukiati B, Saptawati RT, Rodiansyah A. 2021. The potential of amylase enzyme activity against bacteria isolated from several lakes in East Java, Indonesia. Biodiversitas 21: 42-49. Indonesia is one country that has water resources having an abundance of microbial diversity, but not explored massively. This study aims to measure the amylase activity quantitatively from 53 amylolytic bacterial isolates from Ranu Pani, Ranu Regulo, Ranu Grati, and Ngebel Lake; also it identifies the isolate with the highest amylase enzyme activity. The amylase enzyme activity test calculates with DNS (Dinitrosalycylic acid) method, molecular identification of the highest bacterial isolate is based on the 16S rRNA gene. Its relationship is determined through the phylogenetic tree with the Neighbor-Joining (NJ) method. The results showed that the fifty-three bacterial isolates have amylase activity about 0.000-0.016 units/mL. The KN bacterial isolate from Ranu Ngebel was the highest amylase activity, producing enzyme around 0.016 units/mL, while isolate G20 from Ranu Grati was the lowest, reaching about 0.0001 Unit/mL. Based on the morphological and molecular identification, the KN bacterial isolate is classified as the Bacillus cereus group with 99.4-100% sequence similarity, closely related to Bacillus paramycoides (NR 157734.1).

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Aktivitas Enzim Amilase Isolat Bakteri Amilolitik dari Tepung Sagu Basah dan Lingkungan Tempat Penyediaannya Secara Tradisional di Jayapura

JURNAL BIOLOGI PAPUA ◽

10.31957/jbp.457 ◽

2018 ◽

Vol 6 (2) ◽

pp. 47-52

Author(s):

Suprapto Surapto ◽

Tri Gunaedi ◽

Basa T. Rumahorbo

Keyword(s):

Enzyme Activity ◽

Agar Medium ◽

Amylase Activity ◽

Soluble Starch ◽

Bacterial Isolates ◽

Clear Zone ◽

Plate Method ◽

Crude Enzyme Extract ◽

Nutrient Agar Medium ◽

Bacillus Subtilis Atcc

The study about the activity of the enzyme amylase from amylolytic bacterial isolates from wet sagoo starch and its traditional provision environment had been done in Jayapura. The purposes of this study were to determine the activity of amylase enzyme and to identify the bacteria isolated from wet sagoo starch and its processing environment in Jayapura district. The method used was an experimental laboratorium in which isolation of amylolytic bacteria was performed by using nutrient agar medium with 1% soluble starch on spreed pour plate method. The enzyme activity was detected with 0.2% iodine in 2% potassium iodide which were able to form a clear zone. The protein content of the crude enzyme extract was determined by the Bradford method using bovine serum albumin (BSA). Amylase enzyme activity was determined by the formula: DUN/ml = [(R0-R1)/R0] [dilution factor] DUN/ml (dextrinizing units per ml). The results showed that there were 15 isolates amylolytic bacteria. Four (4) bacterial isolates have amylolytic power of more than 30 mm. The amilase activity of amylolytic bacterial of all isolates were quite high: which were 35 577, 18 903, 32 106 and 46 600 U/mg for SU4, SU13, SU23 and SU40 respectively. The identification of isolates indicated that the three isolates are members of the Bacillus cereus ATCC 14 579 types with a similarity value of 71.70% to 81.10%, and one isolate is Bacillus subtilis ATCC 6501 members with a similarity value of 94.30%. Keywords: Amylolytic bacteria, amylase activity, characterization, sago flour.

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Amylolytic ability of bacteria isolated from termite (Coptotermes sp.) gut

Indonesian Journal of Biotechnology ◽

10.22146/ijbiotech.32445 ◽

2018 ◽

Vol 23 (1) ◽

pp. 14 ◽

Cited By ~ 1

Author(s):

Putri Dwi Mulyani ◽

Radhiyah Mardhiyah Hamid ◽

Rifqi Zahroh Janatunaim ◽

Yekti Asih Purwestri

Keyword(s):

Amylase Activity ◽

Rrna Gene ◽

Optimum Temperature ◽

Bacterial Isolates ◽

16S Rrna Gene Sequences ◽

Clear Zone ◽

Optimum Ph ◽

Iodine Test ◽

Brevibacillus Parabrevis

BSR 2, BSR 3, BSR 8, and BSR 9, different bacteria isolated from the termite gut, have been shown to possess cellulolytic activities, but their amylolytic ability has heretofore been unknown. This study attempted to fill in this knowledge gap. The formation of a clear zone using the iodine test showed that the bacteria were able to produce and secrete amylase. Based on the results, the best cultivation times for strains BSR 2, BSR 3, BSR 8, and BSR 9 were 6, 3, 2, and 2 d, respectively, yielding amylase activities of 2.59 ± 0.13 U/mg, 2.00 ± 0.08 U/mg, 1.67 ± 0.10 U/mg, and 1.55 ± 0.12 U/mg, respectively. BSR 2 had the highest amylase activity compared with the other bacterial isolates. The optimum ph for bacterial amylase activity of BSR 2 was 7.0, and the optimum temperature was 40°C. The molecular characterization of isolates BSR 2, BSR 3, BSR 8, and BSR 9 was based on 16S rRNA gene sequences. Isolates BSR 8 and BSR 9 were thus identified as Brevibacillus parabrevis and Brevibacillus sp. With similarities amounting to 92.48% and 95.91%, while the BSR 3 isolate was identified as Pseudomonas alcaligenes with a similarity of 94.29%, and the BSR 2 isolate could not be identified yet.

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

Jahangirnagar University Journal of Biological Sciences ◽

10.3329/jujbs.v5i2.32511 ◽

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)

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OPTIMASI WAKTU INKUBASI PRODUKSI PROTEASE DAN AMILASE ISOLAT BAKTERI ASAL TERASI IKAN TERI Stolephorus sp.

Jurnal Ilmu dan Teknologi Kelautan Tropis ◽

10.29244/jitkt.v10i3.18840 ◽

2018 ◽

Vol 10 (3) ◽

pp. 719-725

Author(s):

Yulia Oktavia ◽

Shanti Dwita Lestari ◽

Susi Lestari ◽

. Herpandi ◽

Miftahul Jannah

Keyword(s):

Enzyme Activity ◽

Incubation Time ◽

Specific Activity ◽

Amylase Activity ◽

Test Results ◽

Bacterial Isolates ◽

Optimum Time ◽

Time Production ◽

Protein Levels ◽

Two Stages

ABSTRAKPenelitian ini bertujuan untuk menentukan waktu optimum produksi enzim protease dan amilase isolat bakteri asal terasi ikan teri Stolephorus sp. Penelitian ini dilakukan dalam dua tahap yaitu pengukuran pertumbuhan isolat bakteri dan penentuan waktu optimum produksi enzim protease dan amilase. Pengujian yang dilakukan meliputi uji aktivitas enzim protease dan amilase dan kadar protein dari tiap-tiap enzim tersebut. Data hasil pengujian dianalisis secara deskriptif. Ada 4 isolat bakteri, 2 isolat merupakan bakteri penghasil protease yaitu isolat P2 dan P4, dan 2 isolat yang merupakan bakteri penghasil amilase yaitu A2 dan A4. Aktivitas protease optimum terjadi pada jam ke-36 untuk isolat P2 sebesar 0,073 U/mL dengan aktivitas spesifik sebesar 1,632 U/mg dan isolat P4 yaitu 0,057 U/mL dengan aktivitas spesifik 4,91U/mg. Aktivitas amilase optimum terjadi pada jam ke-36 untuk isolat A2 sebesar 0,360 U/mL dengan aktivitas spesifik sebesar 7,73 U/mg dan aktivitas amilase optimum pada isolat A4 sebesar 0,239 U/mL dengan aktivitas spesifik 5,24 U/mg. ABSTRACTThe purpose of this research was to know the optimization of incubation time in the production of protease and amylase by bacterial isolates of anchovy Stolephorus sp. paste origin. This research was conducted in two stages, namely the growth of bacterial isolates measurement and determination the optimum time production of the enzyme protease and amylase. Testing conducted include the test proteases and amylase enzyme activity and protein levels of each of these enzymes. The data will be analyzed test results are descriptive. There are 4 bacterial isolates, where 2 isolates, which is a protease-producing bacteria isolates that is P2 and P4, and 2 isolates, which is the amylase-producing bacteria that is A2 and A4. The activity of the protease optimum occurs at to-36 hours to isolate P2 of 0.073 U/mL with a specific activity of 1.632 U/mg and isolates P4 that is 0.057 U/mL with a specific activity of 4.91 U/mg. Whereas amylase activity, optimum occurs at to-36ohours for A2 isolates of 0.360 U/mL with a specific activity of 7.73 U/mg and activity of amylase optimum on A4 isolates of 0.239 U/mL with a specific activity of 5.224 U/mg.

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Indole-3-acetic acid (IAA) producing Pseudomonas isolates inhibit seed germination and α-amylase activity in durum wheat (Triticum turgidum L.)

Spanish Journal of Agricultural Research ◽

10.5424/sjar/2016141-8859 ◽

2016 ◽

Vol 14 (1) ◽

pp. e0802 ◽

Cited By ~ 12

Author(s):

Samira Tabatabaei ◽

Parviz Ehsanzadeh ◽

Hassan Etesami ◽

Hossein A. Alikhani ◽

Bernard R. Glick

Keyword(s):

Acetic Acid ◽

Seed Germination ◽

Durum Wheat ◽

Triticum Turgidum ◽

Amylase Activity ◽

Inhibitory Effect ◽

Bacterial Isolates ◽

Indole 3 Acetic Acid

The role of plant-associated bacteria in plant physiology and metabolism is well documented, but little has been known about the roles played by Pseudomonas in durum wheat (Triticum turgidum L. var durum) growth and development. An in vitroexperiment was conducted to observe the effect of the inoculation of four indole-3-acetic acid (IAA)-producing Pseudomonas isolates and exogenous IAA on seed germination traits and α-amylase activity of durum wheat. The results showed inoculation with all bacterial isolates led to a decrease in the germination percent, although the extent of the depression varied with the isolate. A significant relationship between concentrations of bacterial IAA and the germination inhibition percent in durum wheat seeds by different bacteria strains was observed. The results of this assay showed the effect of bacterial isolates on α-amylase activity after six and 8 days of inoculation was significant, while effect of these isolates on α-amylase activity after two and 4 days of inoculation was not meaningful. In addition, the exogenously applied IAA displayed a concentration-dependent effect on seed germination attributes and α-amylase activity, consistent with the possibility that the inhibitory effect of bacterial inoculation on seed germination was in consequence of bacteria-produced IAA. Therefore, it may suggested that the inhibitory role of IAA in seed germination and α-amylase activity should be taken into account during the screening of IAA-producing Pseudomonas isolates for durum wheat growth promoting agents.

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