双歧杆菌冻干菌粉制备工艺的研究

张 帆，赵 敏，李来酉，鹿晓菲，王 炎，张 锐

(1.东北林业大学生命科学学院，黑龙江 哈尔滨 150040；2.河北科技师范学院生命科技学院，河北 秦皇岛 066004；3.哈尔滨工业大学基础与交叉科学研究院，黑龙江 哈尔滨 150001)

双歧杆菌冻干菌粉制备工艺的研究

张 帆1,2，赵 敏1,*，李来酉1，鹿晓菲1，王 炎3，张 锐2

(1.东北林业大学生命科学学院，黑龙江 哈尔滨 150040；2.河北科技师范学院生命科技学院，河北 秦皇岛 066004；3.哈尔滨工业大学基础与交叉科学研究院，黑龙江 哈尔滨 150001)

优化双歧杆菌冻干菌粉的制备工艺及检测冻干菌粉在模拟人体胃肠环境中的耐受性和贮存稳定性，采用3种方法制备双歧杆菌冻干菌粉。方法1：双歧杆菌在牛奶还原培养基中的扩大培养液加入甘露醇作为保护剂制备冻干菌粉；方法2：双歧杆菌在PTYG培养基中的扩大培养液经低温离心得到菌泥，向菌泥中加入保护剂甘露醇、增量赋形剂脱脂乳粉制备冻干菌粉；方法3：同方法2得到菌泥，向其中加入保护剂甘露醇、包埋剂明胶及增量赋形剂脱脂乳粉制备冻干菌粉。由方法2得到菌粉活菌数较高，为1.31×1010CFU/g；经模拟胃肠环境处理后，活菌数仍保持在4.19×108CFU/g；对胃酸、胆汁酸具有很高的耐受力。经典加速实验证明由方法2制备的菌粉室温保存1年以上活菌数仍保持在107数量级，具有较好的贮存稳定性。

双歧杆菌；菌泥；冻干保护剂；冻干菌粉；活菌数

Abstract ：This study investigated the preparation of lyophilized powder of Bifidobacterium bifidum and assayed its tolerance to simulated human gastrointestinal environment and stability during storage at 37 ℃. Three methods used to prepare lyophilized powder of Bifidobacterium bifidum were described as follows: Method 1: B. bifidum strain was amplified by culturing in reconstituted milk based medium and mannitol was then added as cryoprotectant to prepare lyophilized powder; Method 2:B. bifidum strain was amplified by culturing in PTYG medium and a bacterial mud was then obtained after hypothermal centrifugation. Finally, mannitol as cryoprotectant was added to the bacterial mud to prepare lyophilized powder; Method 3: A bacterial mud obtained as described in Method 2 was added with mannitol as cryoprotectant, gelatin as embedding medium, skim milk powder as excipient to prepare lyophilized powder. The results indicated that a higher viable count was observed in the lyophilized powder prepared by Method 2, which was up to 1.31 × 1010CFU/g and although exposed to simulated gastrointestinal environment, the lyophilized powder exhibited a viable count of up to 4.19 × 108CFU/g, which suggests that the lyophilized powder has excellent tolerance to gastric acid and bile acid. The classic accelerated experiment proved that the lyophilized power obtained Method 2 had good stability during storage at 37 ℃ and the viable count still kept at a magnitude of 107after storage for up to one year.

Key words：Bifidobacterium bifidum；bacterial mud；cryoprotectant；lyophilized powder；viable count of B. bifidum中图分类号：TS201.1 文献标识码：A 文章编号：1002-6630(2010)15-0221-04

双岐杆菌(Bifidobacterium)是人体肠道中最重要的生理性细菌，发挥着生物屏障、营养、免疫、抗肿瘤及改善人体代谢等多种生理作用，被广泛应用于医疗、保健及食品等领域[1]。……