Enhancement of Amylase Activity by Enzymatic Immobilization on Polymer Matrices
Department
Chemistry
Major
Chemistry
Research Advisor
Ma, Yinfa
Advisor's Department
Chemistry
Funding Source
Missouri S& T Opportunities for Undergraduate Research Experiences (OURE) Program; Environmental Research Center, Department of Chemistry
Abstract
Enzyme immobilization has been achieved by a number of techniques, but none have been reported to improve durability or activity of the enzyme. Indeed, the majority of published techniques either denatures the enzyme or physically entraps it. This study serves as a proof-of-concept of a new, chemical-based attachment to a polymer netting, thereby leaving the enzyme intact and functional while being immobilized. Amylase and starch were chosen as a model enzyme-substrate pair for their well-established background as a classical enzymatic reaction. In addition to this, starch was mildly hydrolyzed by hydrochloric acid and heat to accommodate more triiodide molecules. A strong linear correlation (R2 = 0.9998) was observed between starch concentrations and absorbance at 580 nm. This newly developed technique has a method detection limit (MDL) of 1 μg/L (ppb) with a linear range of 1 ppb - 100 mg/L (ppm) in water matrices. The method was further validated through determinations of alpha-amylase activities. Starch degradation revealed typical enzyme kinetics, and enzymatic activity was found to be highly precise (RSD = 1.3%). In conclusion, these simple modifications to the traditional starch-iodine assay can greatly improve UV-VIS sensitivity toward the complex. Results of the activity and/or durability enhancement are pending at this time.
Biography
Casey is from Lake of the Ozarks, MO where he attended School of the Osage high school. His research career began there where he was involved in a number of studies that resulted in publications and presentations. He continued his research path at Missouri University of Science and Technology as a Chemistry major under the guidance of Dr. Yinfa Ma where he specializes in bioanalytical chemistry in his search for noninvasive, early cancer detection techniques. In his spare time, he owns and operates a development company, Sapentia Development, LLC.
Research Category
Sciences
Presentation Type
Oral Presentation
Document Type
Presentation
Location
Upper Atrium/Hallway
Presentation Date
03 Apr 2013, 9:00 am - 11:45 am
Enhancement of Amylase Activity by Enzymatic Immobilization on Polymer Matrices
Upper Atrium/Hallway
Enzyme immobilization has been achieved by a number of techniques, but none have been reported to improve durability or activity of the enzyme. Indeed, the majority of published techniques either denatures the enzyme or physically entraps it. This study serves as a proof-of-concept of a new, chemical-based attachment to a polymer netting, thereby leaving the enzyme intact and functional while being immobilized. Amylase and starch were chosen as a model enzyme-substrate pair for their well-established background as a classical enzymatic reaction. In addition to this, starch was mildly hydrolyzed by hydrochloric acid and heat to accommodate more triiodide molecules. A strong linear correlation (R2 = 0.9998) was observed between starch concentrations and absorbance at 580 nm. This newly developed technique has a method detection limit (MDL) of 1 μg/L (ppb) with a linear range of 1 ppb - 100 mg/L (ppm) in water matrices. The method was further validated through determinations of alpha-amylase activities. Starch degradation revealed typical enzyme kinetics, and enzymatic activity was found to be highly precise (RSD = 1.3%). In conclusion, these simple modifications to the traditional starch-iodine assay can greatly improve UV-VIS sensitivity toward the complex. Results of the activity and/or durability enhancement are pending at this time.
