Authors: Wei Tang Sufang Zhang Haidong Tan Zongbao K Zhao
Publish Date: 2010/03/09
Volume: 45, Issue: 2, Pages: 121-128
Abstract
The malic enzymeencoding cDNA GQ372891 from the oleaginous yeast Lipomyces starkeyi AS 21560 was isolated which has an 1719bp open reading frame flanked by a 290bp 5′ untranslated sequence and a 92bp 3′ untranslated sequence The proposed gene LsME1 encoded a protein with 572 amino acid residues The protein presented 58 sequence identity with the malic enzymes from Yarrowia lipolytica CLIB122 and Aspergillus fumigatus Af293 The LsME1 gene was cloned into the vector pMALp4x to express a fusion protein MBPLsME1 in Escherichia coli TB1 The fusion protein was purified and then cleaved by Factor Xa to give the recombinant LsME1 This purified enzyme took either NAD+ or NADP+ as the coenzyme but preferred NAD+ The K m values for malic acid NAD+ and NADP+ were 085 ± 005 mM 034 ± 008 mM and 74 ± 032 mM respectively at pH 73
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