What is enzyme engineering and different techniques to engineer enzymes?

Answer 1

Enzyme engineering is when you attempt to change the properties of an enzyme, sometimes even introducing novel properties.

There are a lot of different things you can do.

One is to fuse domains (parts) of different proteins together making an enzyme with additional catalytic activities or regulatory domains/inputs. This is done by cloning the coding sequences appropriately. Usually you have to try several fusing possibilities to be able to get a functional enzyme (and it doesn't always work as you hope)

Another is to change the properties of pre-existing parts in the enzyme. By doing so you could be changing the catalytic activity, the binding affinity, the effect of some inhibitors/activators, pH/temperature dependence, stability, even change the substrate.

There are 2 main ways to do this.

One is like Nicegal said, to introduce randomnly mutations in the enzyme and then select for the mutant that has the desired properties. Usually you allow mutations to occur only in regions of the protein that you know they are involved in the fucntion you are interested, but you could also do completely random mutagenesis.

The other is rational design. In order to this you need the crystal structure of the enzyme or of its domains that you want to change. Then you can use computer programmes to predict which specific mutations could give you the desried result and then you make them with PCR site-directed mutagenesis. You actually get a set of possible mutations, so you try more than one construct, but the number of total constructs is by far much smaller than when you are doing random mutagenesis. The drawback is that computer programmes and available crystal structures are not perfect and you might "lose" very good mutations which would come up with the random mutagenesis approach.

Answer 2

Enzyme engineering is a way/method to engineer (change) the enzyme toward your interest. For example, say you want to engineer enzyme, such as proteases, to have better activity when it's cold. This is what detergent company is doing, so that the enzyme proteases can work well even when it's used in cold cycle (laundry washer cycle). There are many different techniques to engineer an enzyme. Personally, I have been using two techniques called regio-specific mutagenesis and error prone PCR. Essentially, both worked in similar ways by introducing mutation into DNA sequence of the enzyme, hoping these mutations will somewhat change the biology of the enzyme and make it better/more efficient to do whatever their purpose ...

Hope it helps!

Answer 3

See here:

http://www.lsbu.ac.uk/biology/enztech/engineering.html

http://www.che.utexas.edu/georgiou/Research/Enzyme_Evolution.htm