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Amit Anand

MOAC MSc mini projects:

According to the MOAC regulations I am suppose to work on three mini research projects, each of 8 weeks in duration in different departments.

My Project title:

Structure/function studies on the bifunctional enzyme Bacillus brevis acetolactate decarboxylase

My main work will be concentrated around the enzyme ALDC (Alpha-aceto-lactate de-carboxylase)

Main initiator of the project: Prof. David Crout

Acetolactate decarboxylase (ALDC) has the unique ability to decarboxylate both enantiomers of acetolactate to give a single enantiomer of the decarboxylation product, (R)-acetoin. This enzyme has application in brewing industry.

A ten week curing process is required during the brewing of beer to ensure that yeast has time to absorb diacetyl, produced during conventional brewing – otherwise the beer has a characteristic ‘off’ odour and taste, rendering it undrinkable.

Acetolactate decarboxylase (ALDC) allows brewers to bypass the reaction that produces diacetyl by catalysing non-oxidative decarboxylation. It similarly catalyses decarboxylation of many other biologically important substrates like some involved in biosynthetic pathway of the essential branch-chain amino acids valine and isoleucine.

ALDC is a two domain protein whose crystal structure has recently been determined at atomic resolution with a Zn ion identified at a likely catalytic site. Its substrates can be racemic but the product is enantiomerically pure. It has no structural homology to any other protein.

General outline:-

Experimental physical science mini project:

Expected Starting date: 14 march 2005

I will be working with my supervisor Prof Martin Wills in the chemistry department. We will be synthesising a new substrate for the enzyme during the first week. In the later weeks I will be doing enzyme kinetics using NMR and my co supervisor will be Dr. Claudia Blindauer.

Expected ending date: 16 may 2005

Experimental Biological sciences mini project:

Expected starting date: 16 may 2005-01-14

Prof Vilmos Fulop is the group leader for ALDC research. I will be working with him to co-crystallize the enzyme with the new substrate under controlled conditions to see the complex at atomic level using X-ray crystallography.

Expected ending date: 11 July 2005

Experimental Computer sciences mini project:

Expected starting date: 11 July 2005

Here I will be working with Prof. Mark Rodger. We will be characterising and modelling the active sites of the enzyme and the substrates. Main aim of the modelling will be to predict more substrates and inhibitors for the enzyme. State of art facilities is available at Centre for Scientific Computing (CSC) for this project. The software that we could use is CHARMm and or Gaussian98.

Expected ending date: 5 September 2005

Reference: Najmudin, S., Andersen, J.T., Patkar, S.A., Borchert, T.V., Crout, D.H.G. & Fülöp, V. (2003). Purification, crystallisation and preliminary X-ray crystallographic studies on acetolactate decarboxylase. Acta Cryst. D59, 1073-1075.





Amit graduated from MOAC after completing the Mathematical Biology and Biophysical Chemistry MSc in 2004/05

He is currently working with the Wilmanns Group, EMBL Hamburg.