Based on the specific region of γ-cyclodextrin glycosyltransferase (γ-CGTase), a gene was explored from the NCBI database by sequence alignment and biological information analysis. The gene was cloned and expressed, and a recombinant enzyme was obtained. The reaction product was identified by high performance liquid chromatography-mass spectrometry (HPLC-MS), which indicated that the enzyme was a γ-CGTase. The molecular weight of the recombinant enzyme was about 80 kDa, the optimum temperature was 50 ℃, and the optimum pH was 10.0. There was no detectable α-cyclodextrin production. The enzyme activity was activated by methanol, ethanol, cyclohexane and dodecanol, but was inhibited by isopropanol and butyl alcohol. The product specificity of the enzyme was affected by the reaction time, the concentration of starch and the volume fraction of ethanol. As indicated by the HPLC analysis result, the proportion of γ-cyclodextrin was increased from 40.11% to 78.20% and the specificity increased by 94.96% in the presence of 10% ethanol. This research could not only provide a novel γ-CGTase but also provide a research basis for changing the product specificity of γ-CGTase.
ZHENG Danni
,
BAI Yuxiang
,
JI Hangyan
,
LI Xiaoxiao
,
WANG Yu
,
JIANG Tong
,
JIN Zhengyu
. Expression, characterization and product specificity of gamma-CGTasefrom Bacillus sp.[J]. Food and Fermentation Industries, 2020
, 46(5)
: 38
-45
.
DOI: 10.13995/j.cnki.11-1802/ts.022791
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