Yati Sodaryati Soeka
| Abstract views: 1960 | PDF views: 1480


Enzymes a-amylase is one of the enzymes used in process of starch degradation. This present study was aimed to characterize and identify of a-amylase producing strain isolated from bulk shrimp-paste in Samarinda, East Kalimantan was carried out. An experiment was conducted to examine influences of incubation, carbon source, substrate concentration temperature for incubation, pH of media, and addition of metal ions. Identification of strain C2 was carried out by using Wizard Genomic DNA Purification Kit (Promega). The result showed that optimum activity of a-amylase from C2 after six days incubation was 18.93 U/mL. Tests on the type of substrate , soluble starch was the best source to produce a- amylase (14.51 U/mL). However, at concentration of 2 % and incubation temperature at 40°C, enzymatic activity was decreased to 12.56 U/mL and 12.79 U/mL, respectively within residual activity of 74.75%. The enzyme activity was 16.43 U/mL and its residual activity was 39.14 % when it was assayed at pH 8.5. Addition of metal ions in the form of divalent and monovalent cations (1 mM) showed that the enzyme could be activated by ion Ca2+ while ion Cu2+, Co2+, Mg2+, Zn2+ , Na+  decrease the activity of the enzyme. Identification of strain C2 using molecular characterization demonstrated that partial sequences of 16S rDNA reffered to as Bacillus subtilis.


shrimp-paste, ?-amylase, enzyme activity, characterization, Bacillus subtilis

Full Text:



Adinarayana K, P Ellaiah and DS Prasad. 2003. Purification and partial characterization of thermostable serine alkaline protease from a newly isolated Bacillus sub-tilis PE-1. American Association of Pharmaceutical Scientists Pharmceutical Sciences Technology 4(4), 440-448.

Adugna S, LAM Alemu, T Kelemu, H Tekola, B Kibret and S Genet. 2004. Medical Biochemistry. Lecture Notes For Health Science Students. In collaboration with the Ethiopia Public Health Training Initiative, The Carter Center, the Ethiopia Ministry of Health, and the Ethio-pia Ministry of Education.

Aehle W. 2004. Enzyme in industry. Production and Applications. 17. Wiley-VCH Verlag GmbH & Co. KgaA, Wein-heim.

Alariya SS, S Sethi, Gupta and BL Gupta. 2013. Amylase activity of a starch degrading bacteria isolated from soil. Archives of Applied Science Research 5(1), 15-24.

Bernfeld P. 1955. Amylases, alpha and beta. In: Methods in Enzymology and Related of Biochemistry Vol. 1. SP Colowick and NO Kaplan (Eds.) 149-152. Academic Press, New York.

Cappuccino JG and N Sherman. 1999. Mikrobiology : A Laboratory Manual, 4th (2,7). Addison- Wesley, Longman. Publishing Company. California USA.

Cecile M. 2011. Impact of calcium on salivary ?-amylase activi-ty, starch paste apparent viscosity and thickness per-ception. Document. Chemosensory Perception 3,112-116.

De Carvalho RV, T Correa and J da Silva. 2008. Properties of an amylase from thermophillic Bacillus sp. Brazillian Journal of Microbiology. 39, 102-107.

Deb P, SA Talukdar, K Mohsina, , PK Sarker, and SMA Sayem. 2013. Production and partial characterization of extracellular amylase enzyme from Bacillus amylo-liquefaciens P-001. SpringerPlus Open Journal 2,154.

El-Tayeb O, F Mohammad, A Hashem and M Aboulwafa. 2007. Optimization of the Industrial Production of Bacterial Alpha Amylase in Egypt IV. Fermentor Production and Characterization of the Enzyme of two Strains of Bacillus subtilis and Bacillus amyloliquefaciens. African Journal of Biotechnology 7(24), 4521-4536.

Fossi BT and Tavea F. 2013. Application of Amylolytic Lactoba-cillus fermentum 04BBA19 in Fermentation for Simul-taneous Production of Thermostable ?-Amylase and Lactic Acid. Chapter 27, 633-658. R & D for Food, Health and Livestock Purposes. Creative Commons License Attribution.

Ginting J. 2009. Isolasi bakteri dan Uji Aktivitas Enzim Ami-lase Kasar Termofilik dari Sumber Air Panas Semangat Gunung Kabupaten Karo, Sumatera Utara [Tesis].

Gurung N, R Sumanta , B Sutapa and R Vivek. 2013. A Broader View: Microbial Enzymes and Their Rele-vance in Industries, Medicine, and Beyond. BioMed Research International. Volume 2013. Article ID 329121.

Hagihara H, K Igarashi, Y Hayashi, K Endo, KI Kitayama, K Ozaki, S Kawai, and S Ito. 2001. Novel ?-Amylase That Is Highly Resistant to Chelating Rea-gents and Chemical Oxidants from the Alkaliphilic Bacillus Isolate KSM-K38. Applied and Environmen-tal Microbiology 67, 1744–1750.

Holtmann G and E Bremer. 2004. RsbV-Independent Induction of the SigB-Dependent General Stress Regulon of Bacillus subtilis during Growth at High Tempera-ture. Journal of Bacteriology 186(18), 6150-6158.

Hoque MM, M Khanam and MA Sheekh. 2006. Characterization and optimization of ?- amylase activity of Streptomyces clavifer. Pakistan Journal of Biological Siences 9 (7), 1328-1332.

Ikram-ul-Haq, H Ashraf, MA Qadeer, J Iqbal. 2005. Pearl millet, a source of alpha amylase production by Bacil-lus licheniformis. Bioresource Technology 96(10), 1201–1204.

Kar S and RC Ray. 2008. Statistical optimization of ?-amylase production by Streptomyces erumpens MTCC7317 cells in calcium alginate beads using response surface methodology. Polish Journal of Microbiology. 57(1), 49-57.

Karata? H, F Uyar, V Tolan, Z Baysal. 2013. Optimization and enhanced production of ?-amylase and protease by a newly isolated Bacillus licheniformis ZB-05 under solidstate fermentation. Annals of Microbiology 63(1), 45-52.

Kiran O, U Comlekcloglu and B Arikan. 2005. Effects of Carbon Sources and Various Chemicals on the Production of a Novel Amylase from Thermophillic Bacillus sp. K-12. Turkish Journal of Biology 29, 99-103.

Mangunwardoyo W, M Takano and I Shibasaki. 1982. Preservation and Utilization of a concentrated seed culture for bacterial amylase production. Annual reports of ICME, vol.5, Osaka University, Osaka, Japan.

Mishra S and N Behera. 2008. Amylase activity of a starch degrading bacteria isolated from soil receiving kitchen wastes. African Journal of Biotechnology 7(18), 3326-3331.

Narang S and T Satyanarayana. 2001. Thermostable ?-amylase production by an extreme thermophile Bacil-lus thermooleovorans. Letters in Applied Microbiology 32(1), 31–35.

Panneerselvam, T. and S. Elavarasi. 2015. Isolation of ?-Amylase Producing Bacillus subtilis from Soil. Interna-tional Journal of Current Microbiology and Applied


  • There are currently no refbacks.