Expression and Activity Assay of Amylomaltase in Pichia pastoris

doi:10.7506/spkx1002-6630-201023057

FOOD SCIENCE ›› 2010, Vol. 31 ›› Issue (23): 258-261.doi: 10.7506/spkx1002-6630-201023057

• Bioengineering • Previous Articles Next Articles

Expression and Activity Assay of Amylomaltase in Pichia pastoris

ZHU Guo-qiang，WANG Shui-xing*，HUANG Lan，ZHU Shi-long

(State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047,China)

Received:2010-08-18 Revised:2010-10-27 Online:2010-12-15 Published:2010-12-29
Contact: WANG Shui-xing E-mail:shuixingw@yahoo.com.cn

Abstract

Abstract:

The gene of MalQ was amplified from genomic DNA of E. coli BL21(DE3) and subcloned intoα secretion signal open reading frame of pPIC9K expression vector to obtain a recombinant plasmid pPIC9K-MalQ. The recombinant plasmid was verified by DNA sequence. The resultant recombinant plasmid bearing MalQ gene was digested by Sal I and transformed into Pichia pastoris strain GS115. The cells with stable expression of amylomaltase were screened in the medium containing G418. The positive clones were induced with methanol to express amylomaltase. The activity of 4-α-glucanotransferase in the crude enzyme was validated through TLC.

Key words: amylomaltase, Pichia pastoris, gene recombination

CLC Number:

Q786

ZHU Guo-qiang，WANG Shui-xing*，HUANG Lan，ZHU Shi-long. Expression and Activity Assay of Amylomaltase in Pichia pastoris[J]. FOOD SCIENCE, 2010, 31(23): 258-261.

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Expression and Activity Assay of Amylomaltase in Pichia pastoris

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