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Xiang Zou,Chang-fa Chen,Xia-chang Qi,Jiang-chao Qian,Ju Chu,Ying-ping Zhuang,Si-liang Zhang,Wei Zeng,Wan-jun Li 한국생물공학회 2010 Biotechnology and Bioprocess Engineering Vol.15 No.6
Improvement of Erythromycin A (Er-A) production and purity by metabolic engineering of the industrial erythromycin-producing strains Saccharopolyspora erythraea strians ZL1004 and ZL1007, in which the amounts of tailoring enzymes EryK (a P450 hydroxylase)and EryG (an S-adenosylmethionine-dependent O-methyltransferase)for biotransformation of Erythromycin D to Er-A were modulated, was performed in a 50 L fermentor. Addition of 15 g/L of corn steep liquor to the medium increased Er-A production; maximum Er-A production was 8,196 U/mL at 191 h, which was 81.8% higher than that of control (4,507 U/mL at 184 h). Er-B impurities were completely eliminated, whereas Er-C impurities were only 153 U/mL at 191 h. Analysis of intra- and extracellular metabolites and key enzyme activities in central carbon metabolism revealed that the pool of TCA cycle intermediates was enhanced by the addition of corn steep liquor and induced an increase in erythromycin biosynthesis. There were no significant differences between strains ZL1004 and ZL1007 regarding Er-A production and impurity accumulation. Compared to wild type strain,Er-A production was improved by 23.9% while Er-C was reduced by 83.9% and Er-B was completely eliminated. Furthermore, fermentation of recombinant strain ZL1004was successfully scaled up from laboratory scale (50 L fermentor) to industrial scale (25 and 132 m3), with similar levels of Er-A production and purity obtained.
Cloning and characterization of the cardiac-specific Lrrc10 promoter
( Xiong Wei Fan ),( Qing Yang ),( You Liang Wang ),( Yan Zhang ),( Jian Wang ),( Jia Jia Yuan ),( Yong Qing Li ),( Yue Qun Wang ),( Yun Deng ),( Wu Zhou Yuan ),( Xiao Yang Mo ),( Yong Qi Wan ),( Karen 생화학분자생물학회(구 한국생화학분자생물학회) 2011 BMB Reports Vol.44 No.2
Leucine-rich repeat containing protein 10 (LRRC10) is characterized as a cardiac-specific gene, suggesting a role in heart development and disease. A severe cardiac morphogenic defect in zebrafish morphants was recently reported but a contradictory result was found in mice, suggesting a more complicated molecular mechanism exists during mouse embryonic development. To elucidate how LRRC10 is regulated, we analyzed the 5`enhancer region approximately 3 kilo bases (kb) upstream of the Lrrc10 start site using luciferase reporter gene assays. Our characterization of the Lrrc10 promoter indicates it possesses complicated cis-and trans-acting elements. We show that GATA4 and MEF2C could both increase transcriptional activity of Lrrc10 promoter individually but that they do not act synergistically, suggesting that there exists a more complex regulation pattern. Surprisingly, knockout of Gata4 and Mef2c binding sites in the 5`enhancer region (-2,894/-2,889) didn`t change the transcriptional activity of the Lrrc10 promoter and the likely GATA4 binding site identified was located in a region only 100 base pair (bp) upstream of the promoter. Our data provides insight into the molecular regulation of Lrrc10 expression, which probably also contributes to its tissue-specific expression. [BMB reports 2011; 44(2): 123-128]