Separation and Purification of Protease from Bacillus licheniformis 2709

doi:10.7506/spkx1002-6630-201011031

FOOD SCIENCE ›› 2010, Vol. 31 ›› Issue (11): 141-146.doi: 10.7506/spkx1002-6630-201011031

• Bioengineering • Previous Articles Next Articles

Separation and Purification of Protease from Bacillus licheniformis 2709

MA Yong-qiang1，ZHANG Hao1，YANG Chun-hua1，LIU Ying1，SUN Bing-yu1，ZHANG Yi-fang2，SHI Yan-guo1,*

1. College of Food Engineering, Harbin University of Commerce, Harbin 150076, China；
2. Harbin Hi-Tech Soybean Food Co. Ltd., Harbin 150078, China

Received:2009-09-03 Revised:2010-03-18 Online:2010-06-01 Published:2010-12-29
Contact: SHI Yan-guo E-mail:zhanghaocomeon@126.com

Abstract

Abstract:

A procedure comprising alcohol precipitation, ammonium sulfate precipitation, DEAE ion exchange chromatography and gel filtration chromatography was used to prepare electrophoretically pure alkaline protease from the fermentation supernatant of Bacillus licheniformis 2709. Deamidation degree and alkaline protease activity were evaluated to optimize the procedure. The results indicated that the activity of purified protease was 61069 U/mg; final purification factor was 38.7; recovery rate of activity was 19.3%; and deamidation rate was 20.9%. The enzymological properties of alkaline protease were also studied. The optimal reaction conditions for alkaline protease were pH 10.0 and 50 ℃. After incubation at 40 ℃ for 2 h, the protease remained more than 80% activity and was relatively stable at pH 8 － 11.

Key words: alkaline protease, purification, deamidation

CLC Number:

TQ925.2

MA Yong-qiang1，ZHANG Hao1，YANG Chun-hua1，LIU Ying1，SUN Bing-yu1，ZHANG Yi-fang2，SHI Yan-guo1,*. Separation and Purification of Protease from Bacillus licheniformis 2709[J]. FOOD SCIENCE, 2010, 31(11): 141-146.

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Separation and Purification of Protease from Bacillus licheniformis 2709

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