Isolation and Characterization of Thermophilic Bacteria from Zharkent geothermal hot spring

Cellulase enzymes can be isolated from thermophile bacteria obtained from the hot spring Ie Seuum, Aceh Besar. This research aimed to recover and characterize the isolates morphologically and biochemically followed by determination of the thermophile bacterial isolates potential as cellulolytic enzyme producers,. The sampling method in this research was conducted by a purposive sampling at temperature of 70 oC, 60 oC and 50 oC. Isolation of thermophilic bacteria was carried out on nutrient agar (NA) media. There were four isolates of thermophilic bacteria isolated recovered at 70 oC, five isolates at 60 oC, and seven isolates at 50 oC. Of the 18 isolates obtained, 15 of them were able to produce cellulase enzymes. Cellulase enzyme production can be determined by the presence of clear zones around bacterial colonies on CMC media after addition of 1% congo red drops and wash with 1 M NaCl. The highest five Cellulolytic Index (CI) values were obtained from isolates ISB75; ISB64; ISB52; ISB54; ISB56 that were 1.23; 2.22; 1.39; 1.59; 1.10, respectively. Biochemical tests carried out on 5 isolates with the highest cellulolytic index values showed that the bacterial isolate were suspected to be from the genera of Bacillus sp.

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Short Communication: Isolation and characterization of thermophilic actinobacteria as proteolytic enzyme producer from Ie Seuum Hot Spring, Aceh Besar, Indonesia

Biodiversitas Journal of Biological Diversity ◽

10.13057/biodiv/d201004 ◽

2019 ◽

Vol 20 (10) ◽

Cited By ~ 1

Author(s):

Lenni Fitri ◽

KARTINI AMELIA PUTRI ◽

SUHARTONO ◽

YULIA SARI ISMAIL

Keyword(s):

Proteolytic Enzyme ◽

Sampling Method ◽

Short Communication ◽

Hot Spring ◽

Thermophilic Bacteria ◽

Skim Milk ◽

Morphological Characterization ◽

Peptide Bonds ◽

Isolation And Characterization

Abstract. Fitri L, Putri KA, Suhartono, Ismail YS. 2019. Short Communication: Isolation and characterization of thermophilic actinobacteria as proteolytic enzyme producer from Ie Seuum Hot Spring, Aceh Besar, Indonesia. Biodiversitas 20: 2802-2808. Proteases are enzymes who catalyze the breakdown of peptide bonds in proteins. This enzyme could be produced from thermophilic bacteria that are able to grow at temperatures of 45-80ºC and are stable to heat. The samples were collected at Ie Seu'um hot spring, Aceh Besar. This study aimed to isolate, to characterize and to determine actinobacteria that were capable of producing protease enzymes. The sampling method in this study was conducted by purposive sampling at temperatures of 50, 60 and 70ºC. Isolation of thermophilic actinobacteria was carried out in Humic Acid Vitamin B (HV) Agar medium and morphological characterization was carried out in Yeast Malt Agar (YMA), Yeast Starch Agar (YSA), Oatmeal Agar (OA) media. Microscopic characterization and measurement of clear zone diameter formed were carried out in skim milk medium. The results showed that one actinobacteria isolate was obtained at a temperature of 50ºC and five isolates at a temperature of 60ºC, meanwhile, no actinobacteria could be obtained at a temperature of 70ºC. A total of 4 isolates obtained were able to produce protease enzymes. The highest Proteolytic Index (IP) value was obtained from IS01 which was 3.8.

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PRODUCTION AND CHARACTERIZATION OF CHITINASE ENZYMES FROM SULILI HOT SPRING IN SOUTH SULAWESI, Bacillus sp. HSA,3-1a

Indonesian Journal of Chemistry ◽

10.22146/ijc.21470 ◽

2010 ◽

Vol 10 (2) ◽

pp. 256-260 ◽

Cited By ~ 6

Author(s):

Hasnah Natsir ◽

Abd. Rauf Patong ◽

Maggy Thenawidjaja Suhartono ◽

Ahyar Ahmad

Keyword(s):

Hot Spring ◽

Thermophilic Bacteria ◽

Extracellular Enzyme ◽

Colloidal Chitin ◽

Bacillus Sp ◽

Optimum Temperature ◽

South Sulawesi ◽

Sds Page ◽

Physiological Analysis

Chitinase is an extracellular enzyme which is capable in hydrolyzing insoluble chitin to its oligomeric and monomeric components. The enzyme produced by thermophilic bacteria was screened and isolated from Sulili hot spring in Pinrang, South Sulawesi, Indonesia. The gram positive spore forming rod shape bacteria was identified as Bacillus sp. HSA,3-1a through morphological and physiological analysis. The production of chitinase enzyme was conducted at various concentration of colloidal chitin at a pH of 7.0 and a temperature of 55 °C. The bacteria optimally was produced the enzyme at a colloidal chitin concentration of 0.5% after 72 h of incubation. The optimum temperature, pH and substrate concentration of chitinase were 60 °C, 7.0 and 0.3%, respectively. The enzyme was stable at a pH of 7.0 and a temperature of 60 °C after 2 h of incubation. The chitinase activities was increased by addition of 1 mM Mg2+, Ca2+ and Mn2+ ions, whereas the activities were decreased by 1 mM Co2+, Fe2+ and Zn2 ions. The molecular weight of the crude enzyme was determined using SDS-PAGE analysis. Five protein fractions were obtained from SDS-PAGE, with MWs of 79, 71, 48, 43 and 22 kDa. Keywords: colloidal chitin, thermophilic bacteria, chitinase

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