一种以球形细菌纤维素为载体制备固定化酶的方法技术

本发明专利技术涉及一种以球形细菌纤维素为载体制备固定化酶的方法，包括：将活化的斜面菌种接入种子培养基培养制得液体种子，然后转接至发酵培养基中，２５～３０℃，８０～２００ｒｐｍ条件下培养４８～９６ｈ，倾滤，得细菌纤维素小球；将细菌纤维素小球用０．１％的ＮａＯＨ在８０～９０℃水浴中处理３０～１２０ｍｉｎ，去离子水漂洗，收集，用０．１％的醋酸中和小球内残留的碱液，静置，去离子水漂洗，将小球取出，吸干表面水分，冷冻，然后冷冻干燥后获得纤维素小球；最后采用物理吸附法或吸附～交联法制备固定化酶。本发明专利技术的球形细菌纤维素小球能高效吸附生物酶，最大限度地保持酶的活性，且安全环保，方法简单，易于操作，可控性强。

Method for preparing immobilized enzyme by using spherical bacterial cellulose as carrier

The invention relates to a method for preparing immobilized enzyme carrier with spherical bacterial cellulose including: slant strain access seed activation medium to prepare the liquid seeds, then transferred to the fermentation medium, 25 to 30 DEG C, 80 ~ 200rpm cultured under 48 ~ 96h, tilt filter, bacteria cellulose beads; bacterial cellulose beads with 0.1% NaOH at 80 to 90 DEG C water bath treatment of 30 ~ 120min, deionized water rinsing, collect, residue with 0.1% acetic acid pellets in lye, static, deionized water rinsing, the ball out, dry surface water, freezing, freezing and drying to obtain cellulose the ball; finally by physical adsorption method or adsorption to prepared immobilized enzyme. The spherical bacterial cellulose ball of the invention can adsorb biological enzyme efficiently, maintain the activity of the enzyme to the maximum, and is safe and environment-friendly. The method is simple, easy to operate and controllable.

【技术实现步骤摘要】

【技术保护点】
一种以球形细菌纤维素为载体制备固定化酶的方法，包括：　　（１）细菌纤维素小球的制备　　将活化的斜面菌种接入ｐＨ４．８～５．２的种子培养基，在２５～３０℃、１００～２００ｒｐｍ条件下培养１２～１６ｈ制得液体种子，以体积百分比为４～１０％接种量将液体种子转接至ｐＨ４．８～５．２的发酵培养基中，每５００ｍＬ锥形瓶中含有２００～３００ｍＬ发酵培养液，２５～３０℃，８０～２００ｒｐｍ条件下培养４８～９６ｈ，倾滤，得细菌纤维素小球；　　（２）细菌纤维素小球的处理　　将发酵收集得到的细菌纤维素小球用质量分数为０．１％的ＮａＯＨ在８０～９０℃水浴中处理３０～１２０ｍｉｎ，用去离子水漂洗３～５次，收集，用体积分...

【技术特征摘要】

【专利技术属性】
技术研发人员：洪枫，邹敏，杨光，杨雪霞，曹张军，杜明，
申请(专利权)人：东华大学，
类型：发明
国别省市：31[]