Puji Lestari, Nur Richana, Rosmimik Rosmimik
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The limited stability of enzyme during long-term storage attributes to its reduced function. In this study, a-amylase from Bacillus licheniformis TVII.6 were formulated with different kind of additives for storage stabilization and better performance. Simultaniously, some minerals and calcium ion were applied to elucidate the inhibition and activation effects to a-amylase. Crude enzyme which was prepared by aceton precipitation was used for this stability test. It demosntrated that 10% of mannitol in citrate phosphate buffer gave the highest residual activity after 3 months of storage (98.5%). Calcium ion influenced the thermal stability of a-amylase and it gave optimum activity at 5 mM CaCl , thus the stability increased from 76.0%/90 C/2 hours to 114.8%/90 C/22 hours in comparison without calcium ions. Calcium ions (5 mM CaCl ) on the stability of a-amylase at 4 C also produced the 2highest residual activity, which remained 100% during 48 hours of incubation. Chemical compounds like FeSO , Na CO and EDTA 4 2 3acted as inhibitors, while (NH ) SO , CuSO , CoSO , MgCl and K HPO did not inhibit activity of a-amylase. NaOH and MnCl 4 2 4 4 4 2 2 4 2 compounds at low concentrations (2 mM) did not inhibit the enzyme but at 10 mM became a-amylase inhibitors. This amylase stability information is very important as a consideration when applying and storing the enzyme, thereby reducing the degradation a-amylase activity.


Additives, enzyme, a-amilase, Bacillus licheniformis TVII.6

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Bernfeld P. 1955. Amylase a and b. In: Methods in Enzymology and Related of Biochemistry, 149-155. SP Colowick and NO Kaplan (Eds.). Acad. Press. New York.

Blanchard JS. 1984. Buffers for enzyme. Meth. Enzymol. 104, 404-414. Bollag DM and SJ Edelstein. 1991. Protein Methods. John Wiley and Sons. New York.

Bose K and D Das. 1996. Thermostable alpha-amylase production using Bacillus licheniformis NRRL B14368. Indian J. Exp. Biol. 34, 1279-1282.

Costa SA, T Tzanov, AF Carneiro, A Paar, GM Gubitz and A Cavaco-Paulo. 2002. Studies of stabilization of native catalase using additives. Enzyme and Microbial Technol. 30, 387-391.

Damardjati DS, U Murdiyatmo, N Richana, Pujoyuwono, N Azizah, D Andriani, P Lestina dan D Kusdiningsih. 1997. Kloning gen-gen amilase dari isolat bakteri indigenous untuk proses biokonversi bahan berpati. Laporan Hasil Penelitian. Balai Penelitian Bioteknologi Tanaman Pangan. Bogor.

Farez-Vidal ME, A Fernandez-Vivas, F Gonzales and JM Arias. 1995. Properties and significance of an a-amylase produced by Myxococcus coralloides D. J.Appl. Bacteriol. 78, 14-19.

Freer SN. 1993. Purification and characterization of the extracellular a-amylase from Streptococcus bovis JB1.Appl. Environ. Microbiol. 59, 1398-1402.

Gupta R, P Gigras, H Mohapatra, VK Goswami, and B Chauhan. 2003. Microbial á-amylases: a biotechnological perspective. Process Biochem. 38, 1599-1616.

Ivanova VN, EP Dobreva, and EI Emanuilova. 1993. Purification and characterization of a thermostable alpha-amylase from Bacillus licheniformis. J. Biotechnol. 28, 277-289.

Kennedy JF, VM Cabalde, and CA White. 1988. Enzymic starch utilization and genetic engineering. Tib. Tech. 6, 84-189.

Kim TV, BG Gu, JY Jeong, SM Byun, and YC Shin. 1995. Purification and characterization of a maltotetraose-forming alkalin a-amylase from an alkalophilic Bacillus Strain GM 8901. Appl. Environ. Microbiol. 61, 3105-3112.

Lee, S., H. Oneda, M. Minoda, A.Tanaka, and K. Inouye. 2006. Comparison of starch hydrolysis activity and thermal stability of two Bacillus licheniformis alpha amylases and insights into engineering á -amylase variants active under acidic conditions. J. Biochem. 139, 997-1005.

Lestari P, N Richana, AA Darwis, K Syamsu, U Murdiyatmo. 2011. Purifikasi dan karakterisasi áamylase termostabil dari Bacillus strearothermophilus TII-12. J. Agrobiogen 7, 56-62.

Longo MA and D Combes. 1999. Thermostability of modified enzymes: a detailed study. J. Chem. Technol. Biotechnol.74, 25-32.

Michiaki M, K Koji, and K Kazuo. 1997. Effects of polyols and organic solvents on thermostability of lipase. J.Chem. Technol. Biotechnol.70, 188-192.

Muchtadi D, SD Nurheri, dan A Made. 1992. Enzim dalam Industri Pangan. PAU Pangan dan Gizi, IPB. Bogor.

Negi S and R Banerjee. 2009.Characterization of amylase and protease produced by Aspergillus awamori in a single bioreactor. Food Research International 42, 443-448

Niaz MT ftikhar, R Tabassum,MA Zia, H Saleem, SQ Abbas, and IU Haq.2010. á-Amylase production by Bacillus licheniformis under solid state fermentation conditions and its cross linking with metalosalts to confer thermostability. International J. Agric. Biol. 12, 793-795

Noriko MK, K Miok, and K Tada. 1999. Stabilization of LAscorbic acid by superoxide dismutase and catalase.Bios. Biotechnol. Biochem. 63, 54-57.

Reddy NS, A Nimmagadda, and KRSS Rao. 2003. An overview of the microbial á-amylase family. African J. Biotechnol. 2, 645-648.

Richana N, GM Yusuf, P Lestari, dan DS Damardjati. 1999. Perilaku kultivasi isolat bakteri termofil penghasil a-amilase. J. Mikrobiol. Indo. 4, 35-39.

Stauffer CE.1989. Enzyme Assays for Food Scientist. van Nostrand Reinhold Publ. New York. Ulger C and C Curakoglu. 2001. Alpha amylase production by Bacillus subtilis and B. amyloliquefaciens in different PEG solutions. World J. Microbiol. Biotechnol. 17, 93-94.

Vihinen M and P Mantsala. 1989. Microbial amylolitic enzymes. Critical Review. Biochem. Molec. Biol. 24, 329-418.

Yang W, L Fan, G Chao-hui, and ZG Ying-Jiu. 2009.Characterization of a Novel Mesophilic Bacterial Amylase Secreted by ZW2531-1, a Strain Newly Isolated from Soil. Chem. Res. Chinese Univ. 25, 198-202.


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