A State-of-the-Art Laboratory for Extremophilic Protein Purification

Abstract

Psychrophiles are organisms that thrive below 10C and make up a large fraction of life on Earth. Psychrophilic (cold-adapted) enzymes are notable for their catalytic efficiency, often outperforming their mesophilic and thermophilic (medium and high temperature respectively) cousins. Cold-adapted proteins accomplish this by remaining flexible at low temperatures, affording low energy binding of substrates and release of products. We hypothesize that this flexibilty is accompanied by the ability to bind non-native substrates (reactants) and convert them into novel products with useful biotechnological properties that can, for example, increase enzymatic reaction rates or serve as novel biosensors. However, psychrophilic enzymes by their nature are highly heat-sensitive, and this thermolability shows up notoriously in their low overexpression and purification yields. To overcome this limitation, we are asking the AFOSR DURIP program to equip a state-of-the-art psychrophilic protein expression and purification instrumentation upgrade, consisting of a modern Fast Protein Liquid Chromatography system supported by low temperature cell lysis and protein lyophilization equipment so that yields of these enzymes can be increased to useful levels to realize the biotechological promise of these systems. Experience with state-of-the-art extremophile protein purification will prepare a new generation of students for jobs in pharma, biotech, and chemical industries, and strengthen the U.S. workforce for challenges we face in the 21st century.

Document Details

Document Type
DoD Grant Award
Publication Date
Feb 29, 2024
Source ID
FA95502310147

Entities

People

  • Robert Stanley

Organizations

  • Air Force Office of Scientific Research
  • Temple University
  • United States Air Force

Tags

Fields of Study

  • Biology
  • Engineering

Readers

  • Microbial Pathology
  • Molecular and Cellular Biochemistry
  • Research Science/Academic Research

Technology Areas

  • Biotechnology