Source: UNIV OF MASSACHUSETTS submitted to NRP
ENHANCING ENZYME STABILITY AND SOLUBILITY IN EXTREME ENVIRONMENTS BY INCORPORATION INTO POROUS COAXIAL ELECTROSPUN NANOFIBER
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
AFRI COMPETITIVE GRANT
Reporting Frequency
Annual
Accession No.
1004885
Grant No.
2015-67011-22820
Cumulative Award Amt.
$79,000.00
Proposal No.
2014-01868
Multistate No.
(N/A)
Project Start Date
Nov 15, 2014
Project End Date
Nov 14, 2016
Grant Year
2015
Program Code
[A7101]- AFRI Predoctoral Fellowships
Recipient Organization
UNIV OF MASSACHUSETTS
(N/A)
AMHERST,MA 01003
Performing Department
Food Science
Non Technical Summary
Enzymes enable vast improvements in catalytic efficiency of a number of important food processing reactions. However, native form enzymes often suffer from low activity, stability, and poor miscibility in processing environments. Enzyme immobilization methods can alter properties without direct modification of the enzyme. By creating a material to introduce enzymes into extreme hydrophobic, ionic, or unideal pH environments, enzymes can have increased storage stability and better activity retention at expanded optimal working conditions. A process requiring enzymatic activity in hydrophobic, ionic, and wide pH range environments in the food industry is protease initiated protein hydrolysis (e.g. nucleotide based flavor production, improved emulsification/foaming/gelation properties). Specifically, chymotrypsin modified pre-digested soy hydrosylates enables hydrolysis of peptide bonds for functionalization and improved emulsifying and foaming properties in foods like beverages and baked goods. Currently the rise in enzymatic modifications of proteins explores using protease's high specificity for individual protein substrates. Incorporation of protease into a porous coaxial nanofiber will allow for access between substrate and enzyme, create a reusable and recoverable material, and allow for increased enzyme solubility in non-aqueous environments with sustained stability. The overall goal of this project is to electrospin coaxial porous hollow nanofibers for improving enzyme solubility and stability in food and bioprocessing applications. To demonstrate this system we will stabilize chymotrypsin into an insoluble electrospun polymer nanofiber to enable controlled peptide modification for improved solubility, emulsion, and foaming capabilities in foods. This method explores the benefits nano-scale materials have over traditional immobilization techniques, and can be incorporated into bioprocessing equipment. Nanofibers can further the storage stability and optimum environments of enzymes used in sustainable agriculture and food as this is a novel application of emerging technology that is multidisciplinary, involving enzymology, bioprocessing, material science, and food chemistry that also may potentially reduce processing waste. The development of nanomaterials for agriculture can improve food processing efficiency aimed at feeding future large populations of the world. The target audience will be subsequently informed by presentation of results at both regional and national conferences that accompany publications in peer reviewed journals. The PD will be available for industry and academic demonstrations, as well as participate in science and engineering educational programs. The long-term outcome of this research would support USDA goals for improving food processing technologies, promoting safe applications of nanotechnology in food systems, improving economic and environmental sustainability of domestic food and agricultural processing. This applied research aims to expand the available technology of electrospinning for use in practical ways in which research can be advanced to benefit society by improving cost effectiveness, catalytic efficiency, and accessibility of beneficial peptides in foods.
Animal Health Component
55%
Research Effort Categories
Basic
5%
Applied
55%
Developmental
40%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
50174101000100%
Knowledge Area
501 - New and Improved Food Processing Technologies;

Subject Of Investigation
7410 - General technology;

Field Of Science
1000 - Biochemistry and biophysics;
Goals / Objectives
· Improve catalytic efficiency of food processing techniques· Develop an enzyme immobilization technique to manage the enzyme interaction in optimum and harsh working environments· Apply the benefits of nano-scale materials to food processing materials· Utilize the properties of food contact materials to modify enzyme environment preferences without direct enzyme structure modification· Facilitate mentorship programs by direct training of junior researchers· Present pertinent project results by both oral presentation and publication in peer reviewed journals to refine technical communication
Project Methods
General scientific methods· Electrospinning - high voltage method to produce nanofiberso Coaxial - variation of electrospinning to control final fiber morphology· Fiber characterizationo Morphology - Scanning Electron Microscope, Transmission Electron Microscope, Confocal Laser Scanning Microscope, surface area· Protein characterizationo Protein contento Secondary structure· Activityo In optimum working environmentso Application in non-ideal enzymatic environments (solvents, pH variations, temperature abuse)Unique aspects of methods· Electrospinning - unique in agriculture and food processing· Development of material characterization and proteins on surfaces characterizationEfforts· Formal classroom instruction, course lectureship with curriculum development· Outreacho STEM outreach activities - science fairs, demonstrations for primary schools· Extension - maintenance of communication avenues with the food industry for final material application· Mentorship of junior researchers leading to further research in related fieldsEvaluation· Experimental tests leading to both quantitative and qualitative resultso Protein contento Enzyme activityo Activity retention· Completion of junior researcher milestones including presentations and coursework assistance· Dissemination of key findings at national and regional conferences· Regular collaborative meetings with co-PD's (biweekly)· Two dissertation examinations of PD within two year project timeline· Peer reviewed publications

Progress 11/15/14 to 11/14/16

Outputs
Target Audience:This project served many target audiences over the two year funding period. A graduate student developed both technical, mentorship and communication skills. Two undergraduate researchers assisting with the completion of this project gained hands-on experience working with the graduate student. Countless primary school children benefited from the outreach this project supports as well, as the graduate student has made STEM outreach a priority for this project. The graduate student gained technical writing experience and oral presentation skills while communicating the findings at three national conferences as well as increased professional network by organizing informational interviewsfunded by this project. The enzymes, biocatalytic materials, and nanotechnology scientific communities received science-based knowledge through a peer reviewed journal article (for submission) and conference presentations as a result of this work. They also received advances in new applications for nano-scale enzyme immobilized materials for food processing. The food industry received understanding of new developments and different approaches for value-added processing, and how bioactive packaging can improve food quality while reducing material and additive use. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?• Communication of findings and networking by participation in national conferences (Gordon Research Conference,American Chemical Society,PD Meeting) Increased professional network by organizing informational interviews with industry and government scientists in research and management • Mentorship of junior graduate student and undergraduate students • Developed individualized research projects • Served on Honors College Thesis Committee How have the results been disseminated to communities of interest?• Journal articles published andfor submission (6 total) • Participation in national conferences (national and regional) as well as roundtable discussions What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? · Improved catalytic efficiency of food processing techniques · Developedenzyme immobilization techniques to manage the enzyme interaction in optimum and harsh working environments · Appliedthe benefits of nano-scale materials to food processing materials · Utilized the properties of food contact materials to modify enzyme environment preferences without direct enzyme structure modification · Facilitated mentorship programs by direct training of junior researchers · Presented pertinent project results by both oral presentation and publication in peer reviewed journals to refine technical communication Change in knowledge • Increased understanding of enzymology partnered with nanotechnology • Increased knowledge in food matrix interactions with food contact materials Change in action • Increased implementation of enzyme immobilization techniques in food processing • Increased prevalence of nano-scale materials in food processing Change in condition • Decreased potential for food processing material waste by increasing biocatalytic efficiency of enzymes in food processing

Publications

  • Type: Journal Articles Status: Under Review Year Published: 2016 Citation: DE Wong, SM Andler, C Lincoln, JM Goddard, and JN Talbert. 2016. Oxygen scavenging polymer coating prepared by hydrophobic modification of glucose oxidase. Journal of Coatings Research and Technology. In Review.
  • Type: Journal Articles Status: Submitted Year Published: 2016 Citation: DE Wong, KJ Senecal, and JM Goddard. 2016. Immobilization of chymotrypsin on hierarchical Nylon 6,6 nanofiber influences enzyme performance. Process Biochemistry. In Review.
  • Type: Journal Articles Status: Other Year Published: 2016 Citation: DE Wong and JM Goddard. 2016. Chymotrypsin encapsulation by emulsion electrospinning polycaprolactone/poly(vinyl alcohol) fibers. For Submission.
  • Type: Theses/Dissertations Status: Accepted Year Published: 2016 Citation: DE Wong. "Enzyme stabilization in hierarchical biocatalytic food packaging and processing materials". Department of Food Science. University of Massachusetts, Amherst. 2016


Progress 11/15/14 to 11/14/15

Outputs
Target Audience:This project served many target audiences in the last year. A graduate student developed both technical, mentorship and communication skills. Two undergraduate researchers assisting with the completion of this projectgained hands-on experience working with the graduate student. Countless primary school children benefitedfrom the outreach this project supports as well, as the graduate student has made STEM outreach a priority for this project. The graduate student gained technical writing experience and oral presentation skills while communicating the findings at three national conferences funded by this project. The enzymes, biocatalytic materials, and nanotechnology scientific communities receivedscience-based knowledge through a peer reviewed journal article (for submission) and conference presentations as a result of this work. They also received advances in new applications for nano-scale enzyme immobilized materials for food processing. The food industry received understanding of new developments and different approaches for value-added processing, and how bioactive packaging can improve food quality while reducing material and additive use. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Communication of findings and networking by participation in national conferences Gordon Research Conference American Chemical Society PD Meeting Mentorship of junior graduate student and undergraduate students Developed individualized research projects Served on Honors College Thesis Committee How have the results been disseminated to communities of interest? Journal article for submission Participation in national conferences Gordon Research Conference American Chemical Society What do you plan to do during the next reporting period to accomplish the goals? Continue to complete specific aims of project outline by applying nanotechnology to agriculture and food processing Participatein national conferences Continue mentorship of junior graduate students and undergraduate students Contribute to department curriculum as course lecturer Submit additional findings to peer reviewed journals Defend dissertation findings

Impacts
What was accomplished under these goals? Change in knowledge Increase understanding of enzymology partnered with nanotechnology Increase knowledge in food matrix interactions with food contact materials Change in action Increase implementation of enzyme immobilization techniques in food processing Increase prevalence of nano-scale materials in food processing Change in condition Decrease potential for food processing material waste by increasing biocatalytic efficiency of enzymes in food processing

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: TM Hancock, DE Wong, and JM Goddard. Dextran as a Cross-Linking Alternative for Immobilized Enzymes. Commonwealth Honors College First Year Research. December 10, 2014. University of Massachusetts, Amherst, MA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: DE Wong, KJ Senecal, and JM Goddard.  Immobilization of chymotrypsin on electrospun nylon 6,6 nanofibers for increased thermostability and activity retention. Gordon Research Conference: Nanoscale Science & Engineering for Agriculture & Food Systems. June 8-11, 2015. Bentley University, Waltham, MA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: DE Wong, KJ Senecal, and JM Goddard. Biocatalytic polymer nanofibers for enzyme immobilization to increase thermostability, pH working range, and activity retention. USDA AFRI NIFA Fellowship Program Project Directors Meeting. August 4-5, 2015. United States Department of Agriculture:NIFA Headquarters, Waterfront Center, Washington DC.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: DE Wong and JM Goddard. Carbohydrate aldehydes as homobifunctional cross-linker analogues for biopolymer stabilization and immobilized enzyme systems. Division of Environmental Chemistry. American Chemical Society 250th National Meeting & Exposition. August 19, 2015. Boston Park Plaza Hotel and Towers, Boston, MA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: DE Wong, KJ Senecal, and JM Goddard. Chymotrypsin immobilized onto surface functionalized macro and nanoscale Nylon 6,6 solid supports. Division of Colloid and Surface Chemistry. American Chemical Society 250th National Meeting & Exposition. August 19, 2015. Boston Convention & Exhibition Center, Boston, MA.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: DE Wong and JM Goddard. Sugar dialdehydes as glutaraldehyde analogues for cross-linked and immobilized chymotrypsin. Division of Agricultural and Food Chemistry. American Chemical Society 250th National Meeting & Exposition. August 20, 2015. Boston Convention & Exhibition Center, Boston, MA.
  • Type: Journal Articles Status: Other Year Published: 2015 Citation: DE Wong, KJ Senecal, and JM Goddard. 2015. Biocatalytic polymer nanofibers for enzyme immobilization to increase thermostability, pH working range, and activity retention. For Submission.