食品科学

• 生物工程 • 上一篇    下一篇

双歧杆菌优良菌株的筛选及耐消化道逆环境性能

熊 涛,黄巧芬,杜 曼   

  1. 南昌大学生命科学与食品工程学院,食品科学与技术国家重点实验室,江西 南昌 330047
  • 出版日期:2014-07-15 发布日期:2014-07-18

Screening of Bifidobacteria with Acquired Excellent Tolerance to Human Gastrointestinal Tract

XIONG Tao, HUANG Qiao-fen, DU Man   

  1. State Key Laboratory of Food Science and Technology, School of Life Sciences and Food Engineering, Nanchang University,Nanchang 330047, China
  • Online:2014-07-15 Published:2014-07-18

摘要:

以改良MRS培养基作为选择性培养基,初筛得75 株严格厌氧且菌落呈蓝色的双歧杆菌疑似菌株。在pH3.0和含3.0 mg/mL胆盐的改良MRS培养基中培养20 h,得到吸光度较高的11 株,并选出耐酸耐胆盐较好的NCU712、NCU701、NCU708进行生理生化鉴定,NCU712基本符合长双歧杆菌的特征。经16S rDNA分子生物学鉴定菌株NCU712为长双歧杆菌。并在模拟人体消化道逆环境下对菌株NCU712的耐受力进行初步研究。结果表明:在pH 3.5以上的人工胃液中,NCU712活菌数随着作用时间的延长而略有上升;pH 2.5时菌株NCU712的D值为30.42 min;pH 3.0时D值为92.97 min,作用2.5 h活菌数保持在107 CFU/mL以上;人工肠液作用4.0 h后,活菌数保持在108 CFU/mL以上;3.0 mg/mL胆盐中培养24 h后,活菌数下降1(lg(CFU/mL))。说明NCU712对人体胃肠道耐受性良好。

关键词: 双歧杆菌, 筛选鉴定, 消化道逆环境, 耐受能力

Abstract:

Totally 75 suspected bifidobacterium were obtained in modified MRSC broth with 60 mg/mL X-Gal. Of these
strains, 11 with high viability were screened by simulating the micro-environment of the human gastrointestinal tract using
MRSC broth (pH 3.0) and then MRSC with 3 mg/mL bile salt for 20 h. Physiological and biochemical tests were conducted
on NCU712, NCU701, and NCU708. They were all identified as B. longum via 16S rDNA sequencing. NCU712 was in line
with the characteristics of B. longum. Preliminary study on its tolerance to the human digestive environment was conducted.
The results showed that the viable count rose slightly with increasing incubation time when the pH of artificial gastric
fluid equaled or exceeded 3.5. The death value (the time it takes to kill 90% of cells) of NCU712 was 92.97 min and 30.42
min in artificial gastric fluid at pH 2.5 and pH 3.0, respectively. The number of viable NCU712 cells remained more than
107 CFU/mL after 2.5 h. The number of living bacterium remained above 108 CFU/mL after 4.0 h in artificial intestinal fluid.
The number of viable NCU712 cells was reduced by one order of magnitude after being incubated in MRSC with 3 mg/mL
bile salt for 24 h. In summary, B. longum NCU712 has excellent tolerance to the human gastrointestinal tract and may be
regarded as functional probiotic with great application potential.

Key words: Bifidobacterium, screening and identification, gastrointestinal tract, tolerance