Lactobacillus fermentum AR497 that can evidently relieve intestinal inflammation in mice was used as the main subject. Based on the Plackett-Burman tests, glycine concentrations, OD600, electric field strength, and sorbitol concentrations were selected as independent variables, the electroporation efficiency was the response value. Further optimization was made by using Box-Behnken response surface methodology. The optimum values for electric field strength,sorbitol content,OD600, and glycine content were 15 kV/cm,0.6 mol/L,0.2 and 26 g/L, respectively. The transformants reached 2.78×105 CFU/μg DNA under the above conditions, which was improved about 22-fold compared with that of unoptimized condition. The highly frequent genetic transformation of AR497 was realized in this work, which laid the foundation for its genetic editing and elucidating its anti-inflammation mechanism.
WEI Yunying
,
ZHAO Yan
,
ZHUANG Jinwei
,
LI Jiang
,
Pamila NAIZEMUDING
,
LU Yisheng
,
AI Lianzhong
,
XIONG Zhiqiang
. Optimization of electroporation method of Lactobacillus fermentum AR497 using response surface methodology[J]. Food and Fermentation Industries, 2019
, 45(7)
: 52
-58
.
DOI: 10.13995/j.cnki.11-1802/ts.018922
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