Document Type

Honors Capstone Project

Date of Submission

Spring 5-1-2012

Capstone Advisor

Professor Robert P. Doyle

Honors Reader

Professor James Hougland

Capstone Major

Biology

Capstone College

Arts and Science

Audio/Visual Component

no

Capstone Prize Winner

no

Won Capstone Funding

yes

Honors Categories

Sciences and Engineering

Subject Categories

Biochemistry | Biochemistry, Biophysics, and Structural Biology

Abstract

Coenzyme Q10 (CoQ10) plays a critical role in energy production in humans. Low levels of CoQ10 have been linked to diseases such as Parkinson's and Huntingtons's disease. CoQ10 levels decrease as we age, and use of statin drugs also lowers CoQ10 levels. Oral supplementation however increases CoQ10 levels. However, the lack of industrial sources of CoQ10 and the difficulty in CoQ10 purification has resulted the need to address these problems. Sporidiobolus johnsonii (S. johnsonii) has been reported as a natural producer of CoQ10. This work is aim to increase the production of CoQ10 in S. johnsonii through genetic engineering. Current purification methods for CoQ10 are difficult and expensive. A protein based purification method may alleviate the current problems associated with typical LC purification. Saposin B (Sap B) has been shown to bind with CoQ10 at selective pH's. We hypothesized that utilizing a Sap B coated support resin that an affinity purification method for CoQ10 could e produced. To this end, the recombinant Sap B was expressed and bound via a HIS-Tag to a sepharose IMAC bead. This work demonstrates that CoQ10 can be bound and released by a Sap B based affinity resin by manipulating the pH.

Creative Commons License

Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License.

Included in

Biochemistry Commons

Share

COinS
 
 

To view the content in your browser, please download Adobe Reader or, alternately,
you may Download the file to your hard drive.

NOTE: The latest versions of Adobe Reader do not support viewing PDF files within Firefox on Mac OS and if you are using a modern (Intel) Mac, there is no official plugin for viewing PDF files within the browser window.