Optimization of Ultrasonic Treatment for Recovery of Recombinant Protein from Pichia pastoris

doi:10.7506/spkx1002-6630-201210012

FOOD SCIENCE ›› 2012, Vol. 33 ›› Issue (10): 57-60.doi: 10.7506/spkx1002-6630-201210012

• Processing Technology • Previous Articles Next Articles

Optimization of Ultrasonic Treatment for Recovery of Recombinant Protein from Pichia pastoris

DAI Jia-kun1,2,3，LI Yan4，MA Qi1,2，GUAN Zheng-jun3，ZHANG Chao3，ZHANG Yan-ting3， ZHAO Rong-rong3，WEI Ya-hui3,*

(1. Institute of Enzyme Engineering, Shaanxi Academy of Sciences, Xi，an 710600, China ；2. Enzyme Engineering Technology Center of Shaanxi, Xi，an 710600, China ；3. Key Laboratory of Resource Biology and Biotechnology in Western China, College of Life Science, Northwest University, Xi，an 710069, China；4. Shaanxi Provincial Institute of Microbiology, Xi，an 710043, China)

Online:2012-05-25 Published:2012-05-07

Abstract

Abstract: In order to establish an appropriate ultrasonic treatment method for the recovery of recombinant protein from Pichia pastoris, the activity of human-adiponectin recombinant protein was evaluated by Western Blotting analysis. A L9(33) orthogonal array design based on single-factor tests was employed to optimize the conditions for ultrasonic treatment. The results showed that the optimal ultrasonic treatment conditions were ultrasonic power of 450 W, ultrasonic treatment time of 25 min and ultrasonic treatment interval of 10 s. Under the optimal ultrasonic treatment conditions, the breaking rate of cell walls was (67.8 ± 2.1)%. The activity of human-adiponectin recombinant protein could be significantly improved by adding 1 mmol/L of PMSF although the addition of PMSF had no effect on breaking rate of cell walls.

Key words: Pichia pastoris, ultrasonication, human-adiponectin protein, Western Blotting analysis

CLC Number:

Q819

DAIJia-kun,，LIYan，MAQi，GUANZheng-jun，ZHANGChao，ZHANGYan-ting，ZHAORong-rong，WEIYa-hui. Optimization of Ultrasonic Treatment for Recovery of Recombinant Protein from Pichia pastoris[J]. FOOD SCIENCE, 2012, 33(10): 57-60.

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Optimization of Ultrasonic Treatment for Recovery of Recombinant Protein from Pichia pastoris

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