Optimisasi Produksi α-Amilase dari Saccharomycopsis fibuligera R64 dengan Response Surface Method-Central Composite Design (RSM-CCD)

Agus Safari, Ahsanul Chaliqin Gayo, Saadah Diana Rachman, Muhammad Yusuf, Safri Ishmayana

Abstract


α-Amilase (EC 3.2.1.1) is an endoamylase that hydrolyze α-1,4 glycosidic bond in amylose and amylopectin molecules yielding simpler carbohydrate. This enzyme is used to replace starch acid hydrolysis in industrial process. α-Amilase is used in many industrial processes including food, paper, bioethanol, and desizing in textile industries. This enzyme can be found in human, plants and microbes. Most industries use microbes due to their flexibility and require less space and time to culture. In the present study we use Saccharomycopsis fibuligera R64 since this yeast has high amylolytic activity and known as food borne microorganisms. Extrinsic factors are easy to manipulate in the process of enzyme production. The present study intended to determine the optimum condition for α-amylase production by S. fibuligera R64 with agitation speed, pH, and time of incubation as independent variable in the present experiment. The experiment was initiated by rejuvenation of S. fibuligera culture, design of experiment using response surface method, analysis of amylase activity, and determination of protein content. The result of the present study showed that optimum condition for amylase production using S. fibuligera R64 is at pH, agitation speed and incubation time of 3.82, 156 rpm and 49 hours, respectively. The specific activity achieved was 423.8 U/mg based on Fuwa assay.

Keywords


α-amilase; fermentation; optimization; response surface method; Saccharomycopsis fibuligera

Full Text:

PDF

References


Ahmed, S.A., Mostafa, F.A., Helmy, W.A., & Abdel-Naby, M.A. (2017) Improvement of Bacterial - α-amylase Production and Application Using Two Steps Statistical Factorial Design. Biocatalysis and Agricultural Biotechnology. 10, 224–233.

Arzita & Agustien, A. (2013) Potensi Bacillus sp. PA-05 Termofilik Obligat Untuk Produksi Amilase. Prosiding SEMIRATA 2013. 1(1), 85–90.

Clementi, F., Rossi, J., Costamagna, L., & Antonie, E. (1980) Production of Amylase(s) by Schwanniomyces castellii and Endomycopsis fibuligera. Antonie van Leeuwenhoek. 46(4), 399–405.

Dar, G.H., Kamili, A.N., Nazir, R., Bandh, S.A., Jan, T.R., & Chishti, M.Z. (2015) Enhanced production of α-amylase by Penicillium chrysogenum in liquid culture by modifying the process parameters. Microbial Pathogenesis. 88, 10–15.

Dinarvand, M., Malahat, R., & Foroughi, M. (2016) Optimizing culture conditions for production of intra and extracellular inulinase and invertase from Aspergillus niger ATCC 20611 by response surface methodology (RSM). Brazilian Journal of Microbiology. 48, 427–441.

Fuwa, H. (1954) A new method for microdetermination of amylase activity by the use of amylose as the substrate. Journal of Biochemistry. 41(5), 583–603.

Hasan, K., Tirta Ismaya, W., Kardi, I., Andiyana, Y., Kusumawidjaya, S., Ishmayana, S., Subroto, T., & Soemitro, S. (2008) Proteolysis of α-amylase from Saccharomycopsis fibuligera: Characterization of digestion products. Biologia. 63(6), 1044–1050.

Ishmayana, S., Kamara, D.S., Rachman, S.D., Kardi, I., & Fadhlillah, M. (2008) Amylase Production from The Yeast Saccharomycopsis fibuligera and Its Potency for Glucose Production from Raw Starch. In Proceeding of The International Seminar on Chemistry. pp. 688–691.

Ismaya, W.T., Hasan, K., Kardi, I., Zainuri, A., Rahmawaty, R.I., Permanahadi, S., El Viera, B.V., Harinanto, G., Gaffar, S., Natalia, D., Subroto, T., & Soemitro, S. (2013) Chemical Modification of Saccharomycopsis fibuligera R64 α-amylase to Improve Its Stability Against Thermal, Chelator, and Proteolytic Inactivation. Applied Biochemistry and Biotechnology. 170(1), 44–57.

Keharom, S., Mahachai, R., & Chanthai, S. (2016) The Optimization Study of α-Amylase Activity Based on Central Composite Design-Response Surface Methodology by Dinitrosalicylic Acid Method. International Food Research Journal. 23(1), 10–17.

Lowry, O.H., Rosebrough, N.J., Randall, R.J., & Lewis, A. (1951) Protein Measurement with The Folin Phenol Reagent. Journal of Biological Chemistry. 193(1), 265–275.

Natalia, D., Vidilaseris, K., Ismaya, W.T., Puspasari, F., Prawira, I., Hasan, K., Fibriansah, G., Permentier, H.P., Nurachman, Z., Subroto, T., Dijkstra, B.W., & Soemitro, S. (2015) Effect of introducing a disulphide bond between the A and C domains on the activity and stability of Saccharomycopsis fibuligera R64 α-amylase. Journal of Biotechnology. 195, 8–14.

Safari, A., Kamara, D.S., Silalahi, F., Fadhlillah, M., Kardi, I., & Ishmayana, S. (2013) Partial Hydrolysis of Purple Sweet Potato Flour by Amylase from Saccharomycopsis fibuligera and Its Application for Composite Breadmaking. Journal of Microbiology, Biotechnology and Food Sciences. 2(5), 2340–2343.

Safari, A., Ishmayana, S., Aini, S.Q., Rachman, S.D., Yusuf, M., Fadhlillah, M., Wulandari, E. & Idar. (2017). Penggunaan Enzim α-Amilase dari Saccharomycopsis fibuligera R64 untuk Peningkatan Kualitas Roti Komposit Terigu-Ubi Jalar Ungu. Al-Kimia. 5(2), 193-207.

Sánchez, S., Bravo, V., Castro, E., Moya, A.J., & Camacho, F. (1997) The influence of pH and aeration rate on the fermentation of D-xylose by Candida shehatae. Enzyme and Microbial Technology. 21(5), 355–360.

Sandhu, D.K., Vilkhu, K.S., & Soni, S.K. (1987) Production of α-Amylase by Saccharomycopsis fibuligera (Syn. Endomycopsis fibuligera). Journal of Fermentation Technology. 65(4), 387–394.

Sanjivani, F. (2016) Optimisasi Produksi Enzim α-Amilase Dari Saccharomycopsis fibuligera dengan Variasi Agitasi dan pH Menggunakan Response Surface Method. Skripsi. Universitas Padjadjaran. Jatinangor.

Singh, R., Kumar, M., Mittal, A., & Mehta, P.K. (2016) Microbial Enzymes: Industrial Progress in 21st Century. 3Biotech. 6, 174.

Stanbury, P.F., Whitaker, A., & Hall, S.J. (2013) Principles of Fermentation Technology. Butterworth-Heinemann. New York.

Tanyildizi, M.S., Özer, D., & Elibol, M. (2005) Optimization of α-amylase production by Bacillus sp. using response surface methodology. Process Biochemistry. 40(7), 2291–2296.

van der Maarel, M.J.E.C., van der Veen, B., Uitdehaag, J.C.M., Leemhuis, H., & Dijkhuizen, L. (2002) Properties and applications of starch-converting enzymes of the α-amylase family. Journal of Biotechnology. 94(2), 137–155.

Wang, S. & Copeland, L. (2015) Effect of acid hydrolysis on starch structure and functionality: a review. Critical Reviews in Food Science and Nutrition. 55(8), 1081–1097.




DOI: https://doi.org/10.24252/al-kimia.v7i1.8117

Refbacks

  • There are currently no refbacks.


Al-Kimia has been indexed in :

See Google Scholar Profile for Al-Kimia by clicking here.

  Mountain View     

More...

Supported By:

Creative Commons License All Publication by Al-Kimia are licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.

Visitor Number: StatCounter - Free Web Tracker and Counter View Al-Kimia Stats

________________________________________________
Al-Kimia, p-ISSN : 2302-2736, e-ISSN : 2549-9335
Faculty of Science and Technology, Alauddin State Islamic University of Makassar, Indonesia
Jl.H.M.Yasin Limpo No. 36 Samata, Gowa, Sulawesi Selatan
Phone: +6281-355-514-523, Email: al-kimia@uin-alauddin.ac.id