Chemical mutagenesis of Fungi

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Loni
Loni's picture
Chemical mutagenesis of Fungi

I need some information about protocols for chemical mutagenesis.
I work with A. oryzae, biological producer of alfa amilaza.I have to make an attempt to increase its enzime activity by meanof chemical mutagenesis.
There are different chemical agnets, but i do not have any protocols.
Despite it is very important to know which is the cheapest way:)
Are there any spcila agnets only for Fungi?

Thank you very much for your attention!!!!!!

drtaing
drtaing's picture
you don't need to use

you don't need to use chemical mutagen to enhance enzyme production. Try with stimulants such as its substrate - in this case soluble starch. Put a bit of substrate can initiate enzyme formation.

The other way is to use UV light to enhance forming mutants. But you are not sure to get the mutants with high enzyme activity.

Good Luck.

jachmoody
jachmoody's picture
Loni wrote:I need some

Loni wrote:

I need some information about protocols for chemical mutagenesis.
I work with A. oryzae, biological producer of alfa amilaza.I have to make an attempt to increase its enzime activity by meanof chemical mutagenesis.
There are different chemical agnets, but i do not have any protocols.
Despite it is very important to know which is the cheapest way:)
Are there any spcila agnets only for Fungi?

Thank you very much for your attention!!!!!!

Just irradiate a mass of spores collected on a membrane filter or on a petri plate until the survival is a small percent of the original population and you'll get a surviving population including some morphological mutants[with color differences , frequently corellating with physiological mutations] --some of the mutants will produce more and some less of the enzyme you're interested in or if you have an x-ray or gamma ray source to produce the mutations that is also a good way.

jim achmoody

itaylor
itaylor's picture
If you're working with a

If you're working with a specific gene I think a nontargeted (brute force) method to find an up-regulated expression mutant would be very laborious.
If you can increase expression with differing media conditions, it may take a while to find, but is pretty inexpensive unless there is an expensive chemical or antibiotic that is needed.

If you've done a lot of molecular work with your species I would suggest a promoter replacement of your choice (regulatable or constitutive). There are all kinds of options here, but the easiest route is to insert another copy of your gene via plasmid or or genomic insertion.
You can replace the endogenous promoter, but better yet obtain an episomal aspergillus plasmid, clone your gene behind a promoter of your choice, transform and what you have is a clone with an extra copy.

Zumrad
Zumrad's picture
I have to increase enzyme

I have to increase enzyme activity in Appergillus sojae (Polygalacturonase) by Mutagenic method. What I have to do? If some one knows it I would like to ask protocol for that.

Zumrad
Zumrad's picture
Hello, could you solve my

Hello, could you solve my problem?