Abstract
To check the suitability of enzyme entrapped beads for use in pharmaceutical industry, amylase enzyme was entrapped in agar/agarose, polyacrylamide gels and calcium alginate beads. Sodium alginate of 1% concentration was found to be best with respect to immobilization efficiency and calcium alginate beads so obtained were not much susceptible to breakage. When sodium alginate- amylase mixture was added from a height of about 20-30 cm. into CaCl2 solution, size of beads was large at higher alginate concentration due to the increase in the size of droplet formation before entering into CaCl2 solution. Enzyme entrapped polyacrylamide and agar/agarose gels were fragile and could not withstand repeated use whereas enzyme entrapped in large calcium alginate beads was used successfully for 50 cycles for the conversion of starch into product without much damage to the beads under stirring conditions. Amylase preparation was also mixed with urease, lysozyme and coimmobilized in large sized calcium alginate beads. These beads were used for 10 repeated cycles to check the conversion of substrates into their products by their respective enzymes and we concluded that an enzyme or mixture of two or three enzymes can be immobilized in the same large sized calcium alginate beads. This will save the additional cost of bioreactor, manpower, maintenance conditions required for the conversion of one drug into another using enzyme/s entrapped in large sized beads.
Highlights
To check the suitability of enzyme entrapped beads for use in pharmaceutical industry, amylase enzyme was entrapped in agar/agarose, polyacrylamide gels and calcium alginate beads
The beads prepared by entrapment of enzyme amylase in polyacrylamide gel, agar/agarose and calcium alginate were checked for the conversion of starch into maltose for 50 repeated cycles
In case of agar/agarose, polyacrylamide and sodium alginate, the different concentrations were tried to choose the best concentration of gel with respect to immobilization efficiency as well as working efficiency of the beads with respect to diffusion of substrate into the beads (Tables 1 and 2)
Summary
Agarose, bisacrylamide were purchased from Himedia laboratories Ltd. Mumbai, India. Amylase enzyme was immobilized by gel entrapment method in calcium alginate, polyacrylamide, agar/ agarose gels. Different concentrations of sodium alginate (0.5, 1, 2, 3 and 4%) mixed with enzyme solution were added separately from a height of nearly 1-2 cm and 20-30 cm, into excess of CaCl2 solution. The beads prepared by entrapment of enzyme amylase in polyacrylamide gel, agar/agarose and calcium alginate were checked for the conversion of starch into maltose for 50 repeated cycles. A mixture of enzymes (amylase, urease and lysozyme) was entrapped in calcium alginate beads and checked the ability of these enzymes to convert their respective substrates into their products. Urease and lysozyme enzymes were filtered separately, concentrated by ultrafiltration and phosphate buffer of pH 7.0 was added to this preparation to give final concentration of 0.1 M. The ammonia is reacted with Nessler’s reagent to form yellow colouration which is measured at 480 nm[28]
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