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The heat-transfer method (HTM): a new thermal principle to evaluating enzymatic reactions and biocatalysis

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  • Full or part time
    Dr Peeters
  • Application Deadline
    No more applications being accepted
  • Funded PhD Project (European/UK Students Only)
    Funded PhD Project (European/UK Students Only)

Project Description

The Engineering and Materials Research Centre (MMU-EMRC) at Manchester Metropolitan University offers up to five fully funded PhD studentships (RCUK-matched stipend of £14,057 and MMU UK/EU PhD student fees of £4,052, both per annum for 2015/16, plus some research expenses), for an April 2016 start.

For full details of the PhD studentship projects available for an April 2016 start: http://www2.mmu.ac.uk/research/research-study/studentships/engineering-and-materials/

Interested applicants should liaise directly with the PhD project Director of Studies to obtain further details on the project.

PROJECT SUMMARY

Development of a thermal device in order to measure catalytic activities of enzymes. This combines engineering (development of flow cells, pumps, etc) with chemistry/biology (the study of DNA restriction enzymes). The applicant will have the possibility to work with new technology.

PROJECT AIMS AND OBJECTIVES

Because of environmental and economic concerns, there is a continuous drive to study alternative energy sources. Out of all available nature-derived renewable energy sources, the main focus is now on enzymatic biofuel cells (EBFCS) [1]. Enzymes are relatively inexpensive compared to traditional precious metal catalysts, and there are a wide range of enzymes available with superior catalytic activity compared to traditional metals. It is complicated to study enzyme activity; therefore, we propose to use a novel thermal method, the heat-transfer method (HTM), which is more straightforward and low-cost compared to traditional methods. The concept of the HTM is based on the analysis of thermal transport through functional interfaces [2]. The central element through which the heat flux will pass consists of a chip serving as an immobilization platform onto which a functional interface is applied (here: enzymes). The internal temperature of the copper block, T1, is measured by a thermocouple and steered via a proportional-integral-derivative controller (PID controller), connected to a power resistor. The front side of the functionalized chip is exposed to the liquid, where T2 is measured at the solid−liquid interface. To extract the heat-transfer resistance Rth (°C/W) quantitatively, the ratio of the temperature difference ΔT = T1 − T2 and the input power P according to Rth = ΔT/P, is analysed.

As a first proof-of-principle, we will evaluate the catalytic activity of two DNA sequences which only vary in one or two nucleotides. This difference makes one of the sequences a substrate for a restriction enzyme (EcoR1), while the other sequence lacks a recognition site and is not a suitable substrate for digestion with EcoR1 [3]. In previous experiments, HTM discriminated quantitatively between DNA strengths with different lengths [2]. The system we propose here contains a minimum of variables since the reaction products in the solution with catalytic activity are practically identical in chemical content to the components present in the mixture without catalytic activity. Therefore, we suggest that any differences in HTM responses (shorter DNA strand = lower thermal resistance) will be interpreted as the reflection of the catalytic activities. Another important point is that it can be used to study the catalytic activity of restriction enzymes in vivo, when using cell extracts of wild type and cdc13-1 mutant.

Aims
Demonstrate the potential to use the heat-transfer method for the evaluation of enzymatic reactions and biocatalysis under flow conditions. The goals of the project include:

-adjust HTM set up with peristaltic pump and develop new flow cells to study enzymatic activity
-measure activity of restriction enzymes towards similar DNA sequences with HTM
-evaluate restriction enzyme activity under in vivo conditions
-surface functionalization of enzymes and study their catalytic activity

SPECIFIC REQUIREMENTS OF THE PROJECT

This is a multidisciplinary project combining engineering, biology and chemistry. The applicant will have the chance to work with novel technology and will be able to measure the effect of enzymes on DNA lengths by monitoring changes at the solid-liquid interface.

We expect the applicant to have:

-A 2.1 Bsc degree in either Engineering, Chemistry or Biology or better
-Since the project is multidisciplinary, preference will be given to applicants with experience in different disciplines (for instance, an engineer who has experience with DNA/enzymes).
-The candidate needs to have strong practical skills.
-It is crucial that the candidate possesses excellent communication skills so he/she can communicate with different departments and present work at international conferences.
-Ability to push research forward
-Analytical skills: critical evaluation of obtained results
-Motivation to solve complex problems
-Ability to work unsupervised (after adequate training)
-Enthusiastic and self-motivated

STUDENT ELIGIBILITY

Project is only open to Home/EU students only

SUPERVISORY TEAM
Informal enquiries can be made to:

Marloes Peeters

+44 (0)161 247 1450
[email protected]

or



David Sawtell

+44 (0)161 247 4642
[email protected]

HOW TO APPLY
Please quote the studentship reference: EMRC-MP-2016-5-PhD.

Applications should be completed using the Postgraduate Research Degree Application Form - http://www2.mmu.ac.uk/media/mmuacuk/content/documents/research/PGR-application-form.doc

Application Form should be emailed to: [email protected]

PLEASE NOTE that Section 9 of the application should be used to write a personal statement outlining your suitability for the study, what you hope to achieve from the PhD and your research experience to date.

CLOSING DATE
1st February 2016

INTERVIEWS
TBC

Funding Notes

Funded studentships will cover tuition fees at the home/EU rate and an RCUK matched bursary of £14,057 per annum. Fully funded PhD studentships at MMU are only available to home and EU students

References

[1] M.S. Dresselhaus, I.L. Thomas, Nature, 2001, 414, 332-337

[2] B. van Grinsven, K. Eersels, M. Peeters et al., ACS AMI Mater. Interf. 2014, 6, 13309-13318

[3] M.K. Zubko, D. Lyall, Nature Cell Biol. 2006, 8, 734-740.

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