Mahajan R, Gupta V K, Sharma J
Department of Biotechnology, Kurukshetra University, Kurukshetra-136 119, India.
Indian J Pharm Sci. 2010 Mar;72(2):223-8. doi: 10.4103/0250-474X.65010.
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 CaCl(2) solution, size of beads was large at higher alginate concentration due to the increase in the size of droplet formation before entering into CaCl(2) 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.
为检验包埋酶珠在制药工业中的适用性,将淀粉酶包埋于琼脂/琼脂糖、聚丙烯酰胺凝胶和海藻酸钙珠中。发现浓度为1%的海藻酸钠在固定化效率方面最佳,如此得到的海藻酸钙珠不太容易破碎。当海藻酸钠-淀粉酶混合物从约20 - 30厘米的高度加入氯化钙溶液时,由于在进入氯化钙溶液之前液滴形成尺寸增大,在较高海藻酸钠浓度下珠的尺寸较大。包埋酶的聚丙烯酰胺和琼脂/琼脂糖凝胶很脆弱,无法承受反复使用,而包埋在大尺寸海藻酸钙珠中的酶在搅拌条件下成功用于淀粉转化为产物的50个循环,且珠体没有受到太大损坏。淀粉酶制剂还与脲酶、溶菌酶混合并共固定在大尺寸海藻酸钙珠中。这些珠子用于10个重复循环,以检验各自的酶将底物转化为产物的情况,我们得出结论,一种酶或两种或三种酶的混合物可以固定在相同的大尺寸海藻酸钙珠中。这将节省使用包埋在大尺寸珠子中的酶将一种药物转化为另一种药物所需的生物反应器、人力、维护条件等额外成本。