The effect of temperature and ethanol treatment on Bacillus subtilis spore cortex-lytic enzyme activity and structure was studied. The activity of the enzyme was determined using UV-visible spectrophotometry, and the changes of secondary and tertiary structures of the enzyme before and after treatment were analyzed using fluorescence spectroscopy and Fourier transform infrared spectroscopy (FTIR). The results showed that when the culture was treated with 30 and 75% ethanol at 80 ℃ for 20 min, the effect of spores denaturation was the best, and the activity of cortical lyase enzyme was the lowest (24.3±2.73, 20.42±2.17 U/L, respectively). The temperature combining with ethanol treatment reduced the fluorescence intensity of cortical lyase enzyme, and the red shifted 3 nm at 80 ℃. Also, the tertiary structure changed. With the increase of temperature and ethanol concentration, the secondary structure also changed: the amount of α-helix and β-sheet were reduced, and random coils increased significantly. The change of secondary structure was the most significant at 80 ℃ with 75% ethanol, the content was 10.98%±0.03, 25.41%±0.03, 48.24%±0.06 and 15.37%±0.02 respectively. The structures of cortex-lytic enzyme after medium temperature combining with ethanol treatment was changed, resulting in the changes of its activity.
CHEN Xiang
,
ZENG Chaowei
,
SHEN Jin
,
GUO Jiajun
,
ZHANG Zhong
. Effect of medium temperature and ethanol on Bacillus subtilis cortex-lytic enzyme activity and structure[J]. Food and Fermentation Industries, 2019
, 45(23)
: 37
-42
.
DOI: 10.13995/j.cnki.11-1802/ts.022068
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