Thermostable amylase activity produced by thermophilic bacteria isolated from Pulu Hotspring, Central Sulawesi

AbstractAmylase is an enzyme that has been widely used as a biocatalyst in foodand bioethanol industries. The availability of thermostable amylase will further expand the market and extend the shelf life of this enzyme. Amylase is produced by amylolytic bacteria using media with high-costnitrogen sources, such as pepton. Black soldier fly (BSF) is a potential source of amylolitic bacteria since its ability to degrade organic matters rapidly. This research aimedtoexploreamylolitic bacteria from the larvae of BSF with highest amylase activity that can be produced using low-cost media. The screening ofamylase activity was conducted by culturing the bacteria on starch containing media.Bacteria with the highest amylase activity were cultured in liquid media with twodifferentnitrogensources (urea and nitrate). Determinations of the optimum pH and temperature for this enzyme activity were carried out in the pH range 4to 7 and temperature 35to 65 ºC. Three amylase-producing isolates were obtained in this study. M1 isolate which has the highest activity was characterized based oncatalase activity and Gram staining. The results showed that the M1 isolate mightbelong togenus Proteussp. At the optimum condition (45ºC and pH 7), amylase activityin nitrate mediawas0.791U/mL, which was about 18-folds higher than that in ureamedia (0,041U/mL). Thus, amylase isolated from BSF larvae can be classified as a mesophilic enzyme and has the potential to be developed commercially at lower production costs.[Keywords:crude extract enzyme,Proteus sp.,thermostable] AbstrakAmilase merupakan salah satu enzim yangtelah digunakan secara luas sebagai biokatalis dalam industri pangan dan bioetanol.Ketersediaan amilase termostabil akan semakin memperluas pasar dan memperpanjang daya simpan enzim ini. Selama ini, produksi amilase dilakukan dengan memanfaatkan bakteri amilolitik menggunakan media dengan sumber nitrogen yang mahal, misalnya pepton. Black soldier fly (BSF) merupakan sumber bakteri amilolitik yang potential karena BSF memiliki kemampuan mendegradasi bahan organik dengan cepat. Penelitian ini bertujuan untuk mengeksplorasi bakteri amilolitik dengan kemampuan amilase tinggi yang dapat diproduksi menggunakan media yanglebihmurah.Skrining bakteri penghasil amilase dilakukan dengan menumbuhkan bakteri pada media yang mengandung pati. Bakteri dengan aktivitas amilase tertinggi dikulturkan dalam media cair dengan dua sumber nitrogenyang berbeda, yaitu urea dan nitrat. PenentuanpH dan suhu optimum aktivitas enzim ini dilakukan pada rentang pH 4sampai 7 dan suhu 35sampai 65 ºC.Tiga isolat penghasil amilase diperoleh dalam penelitian ini. Isolat M1 yang memiliki aktivitas tertinggi dikarakterisasi berdasarkan uji katalasedan uji pewarnaan Gram. Hasilnya menunjukkan bahwa isolat M1 termasukgenus Proteus sp. Pada kondisi optimum (suhu 45oC dan pH 7), aktivitas amilase pada media nitrat adalah 0,791 U/mL, lebih kurang 18 kali lebih tinggi dibanding aktivitas pada media urea (0,041 U/mL). Dengan demikian, amilase yang dihasilkan oleh bakteri asallarva BSF merupakan enzim mesofilik dan berpotensi untuk dikembangkan secara komersial dengan biaya produksi yang lebih murah.[Kata kunci: enzim ekstrak kasar, Proteussp.,termostabil]

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SKRINING BAKTERI TERMOFILIK POTENSIAL AMILOLITIK DARI SUMBER AIR PANAS WAY BELERANG KALIANDA LAMPUNG SELATAN

Journal of Aquatropica Asia ◽

10.33019/aquatropica.v6i1.2458 ◽

2021 ◽

Vol 6 (1) ◽

pp. 1-7

Author(s):

Sefi Desfeni Mawati ◽

Esti Harpen ◽

Hilma Putri Fidyandini

Keyword(s):

Incubation Temperature ◽

Hot Spring ◽

Thermophilic Bacteria ◽

Pseudomonas Stutzeri ◽

Optimum Temperature ◽

Bacterial Isolates ◽

Inhibitory Zone ◽

Biochemical Tests ◽

Amylolytic Enzyme ◽

Thermostable Amylase

Thermophilic bacteria that produced amylase and protease have been isolated from Way Belerang hot spring, Kalianda, South Lampung. This research aims to screen and identify thermophilic bacteria that have the potential to produce thermostable amylase and protease enzymes.The research procedures included sampling, isolation of enzyme-producing thermophilic bacteria, a series of phenotypic and biochemical tests, and molecular identification by 16s rRNA. This study used 2 treatments, namely incubation temperature 37 and 50 ºC with 3 repetitions. The results showed that the optimum temperature for growth of thermophilic bacterial isolates and thermophilic bacterial isolates producing amylase enzymes was 50ºC. The bacteria isolate that had the best amylolytic enzyme activity was Isolate A.WB.50.1 with a diameter of the inhibitory zone was 15.44 mm. Isolate A.WB.50.1 has been identified by the species Pseudomonas stutzeri.

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Molecular identification of amylase-producing thermophilic bacteria isolated from Bukit Gadang Hot Spring, West Sumatra, Indonesia

Biodiversitas Journal of Biological Diversity ◽

10.13057/biodiv/d210319 ◽

2020 ◽

Vol 21 (3) ◽

Author(s):

Aulia Ardhi ◽

Arina Nadenggan Sidauruk ◽

Nabella Suraya ◽

Nova Wahyu Pratiwi ◽

Usman Pato ◽

...

Keyword(s):

High Temperature ◽

Molecular Identification ◽

Hot Springs ◽

Hot Spring ◽

Thermophilic Bacteria ◽

Amylase Activity ◽

Hydrolytic Enzymes ◽

Molecular Technique ◽

West Sumatra ◽

Pharmaceutical Industries

Abstract. Ardhi A, Sidauruk AN, Suraya N, Pratiwi NW, Pato U, Saryono. 2020. Molecular identification of amylase-producing thermophilic bacteria isolated from Bukit Gadang Hot Spring, West Sumatra, Indonesia. Biodiversitas 21: 994-1000. Amylase is one of the hydrolytic enzymes that is widely used in a wide number of industrial processes such as food, fermentation, textile, paper, detergent, and pharmaceutical industries. Amylase produced by thermophilic bacteria may be thermostable, which is very beneficial in several applications requiring high temperature, for example, the process of gelatinization, liquefaction, and saccharification are performed in high temperature involved in the starch processing. In this study, the amylase-producing ability of thermophilic bacteria isolated from Bukit Gadang hot spring, West Sumatra, Indonesia, was checked and followed by molecular identification. Thirteen isolates that were successfully isolated from the hot springs were microscopically and macroscopically characterized, biochemically tested, and determined their amylase enzyme activity both qualitatively and quantitatively. The isolate that performed the best amylase activity was identified using the molecular technique. The DNA sequencing was carried out in 16S rRNA and continued with BLAST search for species identification. The result of molecular identification showed that the isolate with the best amylase activity was identified as Bacillus licheniformis. The optimum amylase production (231.33 U/ml) and the best enzyme-specific activity (101.79 U/mg) were obtained at the incubation time of 36 hours.

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Isolation of Thermophilic Bacteria from Bora Hot Springs in Central Sulawesi

Biosaintifika Journal of Biology & Biology Education ◽

10.15294/biosaintifika.v10i2.14905 ◽

2018 ◽

Vol 10 (2) ◽

pp. 291-297 ◽

Cited By ~ 1

Author(s):

Slamet Ifandi ◽

Muh Alwi

Keyword(s):

Hot Springs ◽

Hot Spring ◽

Thermophilic Bacteria ◽

Determinative Bacteriology ◽

Thermostable Enzymes ◽

Observational Study Design ◽

Biochemistry Test ◽

Last Stage ◽

Design Characteristics ◽

Central Sulawesi

Thermophilic bacteria can survive at high temperature, in which hot spring is one of its habitats. Indonesia has many hot springs with potential as a habitat for thermophilic bacteria. The purpose of this study was to obtain isolates thermophilic bacteria from Bora hotspring located in Central Sulawesi. This study applied a descriptive-observational study design, characteristics of bacterial properties identified using conventional methods according to the Bergey’s Manual of Determinative Bacteriology. The study was conducted in 3 stages. The first stage was bacteria cultivation on the appropriate media, followed by stage of isolated and the last stage by identified characteristics of thermophilic bacteria which included microscopic and macroscopic morphology, Physiological and biochemistry test. The results of the isolates indicted 4 representative isolated of thermophilic bacteria from Bora Hot Spring namely TM022, TM023, TM024, TM026. The bacteria isolates obtained were bacillus, coccus and Gram negative and positive, while the physiological test of all isolates were able to grow and showed changes in the medium. This study is useful in providing characteristic of indigenous thermophilic bacteria isolates that produces thermostable enzymes.

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The inhibitory effect of the crude extract of Centella asiatica on α-amylase activity

Planta Medica ◽

10.1055/s-0030-1264380 ◽

2010 ◽

Vol 76 (12) ◽

Author(s):

N Supkamonseni ◽

A Thinkratok ◽

R Srisawat

Keyword(s):

Crude Extract ◽

Amylase Activity ◽

Centella Asiatica ◽

Inhibitory Effect

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DAYA SAING DAN PERAN PEMERINTAH DALAM MENINGKATKAN DAYA SAING KOMODITI KAKAO DI SULAWESI TENGAH

Buletin Ilmiah Litbang Perdagangan ◽

10.30908/bilp.v11i1.79 ◽

2017 ◽

Vol 11 (1) ◽

pp. 69-96

Author(s):

Siti Yuliaty Chansa Arfah ◽

Harianto . ◽

Suharno .

Keyword(s):

Policy Analysis ◽

Government Policy ◽

Secondary Data ◽

Research Area ◽

Primary Data ◽

Cocoa Beans ◽

Output Analysis ◽

The Government ◽

Central Sulawesi ◽

Policy Analysis Matrix

Penelitian ini bertujuan untuk mengkaji daya saing komoditi kakao di Sulawesi Tengah dan melihat peran pemerintah dalam meningkatkan daya saing komoditi kakao. Data primer berasal dari observasi, wawancara dan kuesioner, sementara data sekunder berasal dari instansi terkait. Metode analisis menggunakan Policy Analysis Matrix (PAM) dan analisis sensitivitas. Penelitian ini menemukan bahwa nilai PCR Kabupaten Parigi Moutong 0,589 dan Kabupaten Sigi 0,396. Sedangkan nilai DRC Kabupaten Parigi Moutong 0,387 dan Kabupaten Sigi 0,319. Hal tersebut mengindikasikan bahwa usahatani komoditi kakao di Sulawesi Tengah memiliki daya saing, namun tidak menguntungkan secara ekonomi karena Sulawesi Tengah menghasilkan biji kakao yang tidak difermentasi akibatnya petani menerima harga rendah. Berdasarkan hasil analisis, pemerintah belum memberikan proteksi terhadap harga biji kakao dalam negeri melalui harga referensi biji kakao sehingga harga biji kakao didaerah penelitian masih tergolong rendah jika dibandingkan dengan harga di pasar internasional. Sementara terhadap input, pemerintah telah memberikan kebijakan subsidi kepada petani, namun implementasinya masih perlu perbaikan terutama terkait penyaluran dan pengelolaan bantuan agar merata. Kajian ini merekomendasikan masih diperlukan kebijakan pemerintah baik terhadap input maupun output untuk meningkatkan produktivitas, menurunkan biaya produksi dan menaikkan harga jual biji kakao, sehingga dapat meningkatkan daya saing biji kakao. The purpose of this study is to assess the competitiveness of cocoa in Central Sulawesi and to investigate the role of government in improving the competitiveness of cocoa. The primary data were generated through observation, interviews and some questionnaires. The secondary data were obtained from the agency or the institution related to the research. This study uses the Policy Analysis Matrix (PAM) and sensitivity analysis. The study found that the PCR value for Parigi Moutong district was 0.589 and Sigi district was 0.396. While, the DRC value for Parigi Moutong district was 0.387 and Sigi district was 0.319. This indicates that cocoa beans farming in Central Sulawesi has competitiveness, but not economically beneficial because Central Sulawesi produces unfermented cocoa beans consequently farmers receive low prices. Based on the results of the government's impact on output analysis, the government did not provide protection for domestic cocoa seed prices through the reference price of cocoa beans, consequently the price of domestic cocoa beans, particularly in the research area, was relatively low compared to the price of cocoa beans at the international market. Seen from the government policy on inputs, the government have provided subsidies to farmers but they need to improve the distribution and management of aid to be evenly distributed. It is necessary to set up good government policy on inputs and outputs in order to increase cocoa seed productivity, decrease production cost and increase the price which simultaneously can improve its competitiveness in the research location.

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