Expression, Purification and Enzymological Properties of Recombinant Elastase

doi:10.7506/spkx1002-6630-200919052

FOOD SCIENCE ›› 2009, Vol. 30 ›› Issue (19): 227-231.doi: 10.7506/spkx1002-6630-200919052

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Expression, Purification and Enzymological Properties of Recombinant Elastase

GU Xin-xi1，XU Wen-tao2，HUANG Kun-lun2，LUO Yun-bo2，LIN Xi-jin2，CHEN Zhuo-jun2，TIAN Hong-tao1,2,*

1. College of Food Science and Technology, Hebei Agricultural University, Baoding 071001, China ；
2. College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China

Received:2008-10-14 Revised:2009-07-11 Online:2009-10-01 Published:2010-12-29
Contact: TIAN Hong-tao E-mail:xuwentao1111@sina.com;cauxwt@yahoo.cn

Abstract

Abstract:

Elastase was expressed in Pichia pastoris KM7 with the transformation of a recombinant plasmid pPIC3.5K/PAE encoding a full-length lasB gene of PAE (Pseudomonas aeruginosa elastase) under the induction of methanol. The optimal expression conditions were 1% methanol and 4 d induction. The best purification efficiency was achieved through DEAESepharose FF anion exchange chromatography. The optimal pH and temperature range for catalytic reaction of this elastase were 6.0 8.5 and 25 40 , respectively. Its activity was revealed an enhancement effect from Ca2+ and Mg2+, and an inhibition effect from Zn2+, Cd2+, Mn2+, Fe2+, Co2+, Cu2+ and EDTA.

Key words: elastase, induction, purification , enzymatic property

CLC Number:

GU Xin-xi1，XU Wen-tao2，HUANG Kun-lun2，LUO Yun-bo2，LIN Xi-jin2，CHEN Zhuo-jun2，TIAN Hong-tao1,2,*. Expression, Purification and Enzymological Properties of Recombinant Elastase[J]. FOOD SCIENCE, 2009, 30(19): 227-231.

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Expression, Purification and Enzymological Properties of Recombinant Elastase

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