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Chemical modification of saccharomycopsis fibuligera r64 α-amylase to improve its stability against thermal, chelator, and proteolytic inactivation
Ismaya W.T.a,c, Hasan K.a,d, Kardi I.a, Zainuri A.a, Rahmawaty R.I.a, Permanahadi S.a, El Viera B.V.a, Harinanto G.b, Gaffar S.a, Natalia D.b, Subroto T.a, Soemitro S.a
a Department of Chemistry, Faculty of Mathematics and Natural Sciences, Padjadjaran University Jalan, Indonesia
b Biochemistry Research Division, Center for Life Sciences, Bandung Institute of Technology, Indonesia
c Institute for Bioengineering and Biosciences, Georgia Institute of Technology, United States
d Department of Experimental Biology, Research Center for Toxic Compounds in the Environment, Masaryk University, Czech Republic
[vc_row][vc_column][vc_row_inner][vc_column_inner][vc_separator css=”.vc_custom_1624529070653{padding-top: 30px !important;padding-bottom: 30px !important;}”][/vc_column_inner][/vc_row_inner][vc_row_inner layout=”boxed”][vc_column_inner width=”3/4″ css=”.vc_custom_1624695412187{border-right-width: 1px !important;border-right-color: #dddddd !important;border-right-style: solid !important;border-radius: 1px !important;}”][vc_empty_space][megatron_heading title=”Abstract” size=”size-sm” text_align=”text-left”][vc_column_text]α-Amylase catalyzes hydrolysis of starch to oligosaccharides, which are further degraded to simple sugars. The enzyme has been widely used in food and textile industries and recently, in generation of renewable energy. An α-amylase from yeast Saccharomycopsis fibuligera R64 (Sfamy) is active at 50 C and capable of degrading raw starch, making it attractive for the aforementioned applications. To improve its characteristics as well as to provide information for structural study ab initio, the enzyme was chemically modified by acid anhydrides (nonpolar groups), glyoxylic acid (GA) (polar group), dimethyl adipimidate (DMA) (cross-linking), and polyethylene glycol (PEG) (hydrophilization). Introduction of nonpolar groups increased enzyme stability up to 18 times, while modification by a cross-linking agent resulted in protection of the calcium ion, which is essential for enzyme activity and integrity. The hydrophilization with PEG resulted in protection against tryptic digestion. The chemical modification of Sfamy by various modifiers has thereby resulted in improvement of its characteristics and provided systematic information beneficial for structural study of the enzyme. An in silico structural study of the enzyme improved the interpretation of the results. © 2013 Springer Science+Business Media New York.[/vc_column_text][vc_empty_space][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][vc_empty_space][megatron_heading title=”Author keywords” size=”size-sm” text_align=”text-left”][vc_column_text]Chemically modified,Cross linking agents,Enzyme engineering,Polyethylene glycol (PEG),Saccharomycopsis,Structural studies,Structure-function relationship,Tryptic digestion[/vc_column_text][vc_empty_space][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][vc_empty_space][megatron_heading title=”Indexed keywords” size=”size-sm” text_align=”text-left”][vc_column_text]α-Amylase,Chemical modification,Enzyme engineering,Saccharomycopsis fibuligera,Structure-function relationship,Tryptic digestion[/vc_column_text][vc_empty_space][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][vc_empty_space][megatron_heading title=”Funding details” size=”size-sm” text_align=”text-left”][vc_column_text]Acknowledgments The research was financed by the Indonesian Directorate General of Higher Education (DIKTI), Ministry of National Education, Republic of Indonesia (competitive grant batch IX/1-3 entitled Protein engineering of α-amylase from Saccharomycopsis fibuligera R64, for WTI). We thank Prof. J.J. Beintema for his valuable advice and discussion and Dr. H.J. Doddema for editing the manuscript.[/vc_column_text][vc_empty_space][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][vc_empty_space][megatron_heading title=”DOI” size=”size-sm” text_align=”text-left”][vc_column_text]https://doi.org/10.1007/s12010-013-0164-8[/vc_column_text][/vc_column_inner][vc_column_inner width=”1/4″][vc_column_text]Widget Plumx[/vc_column_text][/vc_column_inner][/vc_row_inner][/vc_column][/vc_row][vc_row][vc_column][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][/vc_column][/vc_row]