6/14/18

Dear Vijay,

I once prepared immobilized yeast cells in Ca-Alginate gels and was able to store them w/ viability retention for several days in the refrigerator in 0.1 M K-Acetate buffer @ pH 5.1.

In the case of your enzyme, I would store it in the refrigerator in a buffer that provided the optimum pH.

I hope this information helps you.

Bill Colonna Center for Crops Utilization Research, Iowa State University, Ames, Iowa, USA [email protected]

Dear Sir,

Thank you so much for your reply but here we are facing a problem I.e. when we store in phosphate buffer the cal. alginate beads dissolved in it.

So, please suggest the solution.

Thanks

6/14/18

Dear Vijay,

The PO4 is probably chelating the Ca away from the alginate, causing it to dissolve. Are you seeing a precipitate (calcium PO4) when this happens?

One solution is to use a different (PO4-free) buffer. Is this something you can do? What pH do you need for the enzyme? There are many buffers that may have the same pH range you need for good enzyme activity without causing the CaAlginate to dissolve, e.g., Tris, Bicine, Tricine, MOPS, HEPES, etc. You can do a google search or go on Sigma Chemical's website to find a list of these buffers as well as their pH ranges. If they don't somehow interfere with the assay system or cause problems w/ the alginate, this approach may work for you. I hope this information helps you.

Bill Colonna

The storage and stability of the hydrogel in the buffer depends on various parameters (e.g., degree of cross-linking, concentration and pH of the storage buffer, its type and the storage temperature). Check the following paper, if you want to use PBS to determine these conditions.

Also, 0.05 M HEPES–tris buffer can be used for the storage as in the attached paper.

Dear Vijay

You can use 0.05 M HEPES–tris buffer. I do thing this will work for you, try it.

I do not recommend storage for long time even in buffer as it affect the cross linking....etc

Freshly used alginate gel is highly recommended.

I would not recommend storage of immobilized enzyme