scholarly journals Bioproduction of trans-10,cis-12-Conjugated Linoleic Acid by a Highly Soluble and Conveniently Extracted Linoleic Acid Isomerase and an Extracellularly Expressed Lipase from Recombinant Escherichia coli Strains

2018 ◽  
Vol 28 (5) ◽  
pp. 739-747 ◽  
Author(s):  
Meng nan Huang ◽  
Xin yao Lu ◽  
Hong Zong ◽  
Bin Zhuge ◽  
Wei Shen
Keyword(s):  
Escherichia Coli ◽  
Linoleic Acid ◽  
Conjugated Linoleic Acid ◽  
Recombinant Escherichia Coli ◽  
Linoleic Acid Isomerase

scholarly journals A New Member of the Escherichia coli fad Regulon: Transcriptional Regulation of fadM (ybaW)

2009 ◽  
Vol 191 (20) ◽  
pp. 6320-6328 ◽  
Author(s):  
Youjun Feng ◽  
John E. Cronan
Keyword(s):  
Escherichia Coli ◽  
Fatty Acids ◽  
Linoleic Acid ◽  
Conjugated Linoleic Acid ◽  
Unsaturated Fatty Acids ◽  
Negative Regulator ◽  
Receptor Protein ◽  
Conjugated Linoleic Acid Isomer ◽  
Linoleic Acid Isomer

ABSTRACT Recently, Nie and coworkers (L. Nie, Y. Ren, A. Janakiraman, S. Smith, and H. Schulz, Biochemistry 47:9618-9626, 2008) reported a new Escherichia coli thioesterase encoded by the ybaW gene that cleaves the thioester bonds of inhibitory acyl-coenzyme A (CoA) by-products generated during β-oxidation of certain unsaturated fatty acids. These authors suggested that ybaW expression might be regulated by FadR, the repressor of the fad (fatty acid degradation) regulon. We report mapping of the ybaW promoter and show that ybaW transcription responded to FadR in vivo. Moreover, purified FadR bound to a DNA sequence similar to the canonical FadR binding site located upstream of the ybaW coding sequence and was released from the promoter upon the addition of long-chain acyl-CoA thioesters. We therefore propose the designation fadM in place of ybaW. Although FadR regulation of fadM expression had the pattern typical of fad regulon genes, its modulation by the cyclic AMP (cAMP) receptor protein-cAMP complex (CRP-cAMP) global regulator was the opposite of that normally observed. CRP-cAMP generally acts as an activator of fad gene expression, consistent with the low status of fatty acids as carbon sources. However, glucose growth stimulated fadM expression relative to acetate growth, as did inactivation of CRP-cAMP, indicating that the complex acts as a negative regulator of this gene. The stimulation of fadM expression seen upon deletion of the gene encoding adenylate cyclase (Δcya) was reversed by supplementation of the growth medium with cAMP. Nie and coworkers also reported that growth on a conjugated linoleic acid isomer yields much higher levels of FadM thioesterase activity than does growth on oleic acid. In contrast, we found that the conjugated linoleic acid isomer was only a weak inducer of fadM expression. Although the gene is not essential for growth, the high basal level of fadM expression under diverse growth conditions suggests that the encoded thioesterase has functions in addition to β-oxidation.

Heterologous Expression of Production of T10, C12-CLA Linoleic Acid Isomerase Gene from Propionibacterium acnes

2013 ◽  
Vol 641-642 ◽  
pp. 765-768
Author(s):  
Xue Luo ◽  
Lan Wei Zhang ◽  
Hong Bo Li ◽  
Shu Mei Wang ◽  
Chao Hui Xue
Keyword(s):  
Escherichia Coli ◽  
Linoleic Acid ◽  
Double Bond ◽  
Propionibacterium Acnes ◽  
Expression System ◽  
Fusion Partner ◽  
His Tag ◽  
Double Bond System ◽  
Bond System ◽  
Linoleic Acid Isomerase

The linoleic acid isomerase enzyme from Propionibacterium acnes, which catalyzed the isomerization of a methylene-interrupted double bond system to a conjugated double bond system, creating trans-10, cis-12 conjugated linoleic acid (t10, c12 CLA) was cloned and overexpressed in Escherichia coli using expression system of pCold-SUMO. The gene was expressed under T7 promotor with a fusion partner of 6×His. Tag at its 5'terminal. After induction by 0.2mM IPTG for 18 h, the recombinant enzyme are expressing in the cytoplasm. The 1275 bp gene from P. acnes encode a linoleic acid isomerase protein with 424 amino acids (55KD).

Molecular Cloning and Heterologous Expression of Linoleic Acid Isomerase Gene from Lactobacillus reuteri and Lactobacillus acidophilus

2012 ◽  
Vol 554-556 ◽  
pp. 1410-1414 ◽  
Author(s):  
Xue Luo ◽  
Lan Wei Zhang ◽  
Tie Nan Luo ◽  
Shu Mei Wang
Keyword(s):  
Escherichia Coli ◽  
Amino Acids ◽  
Linoleic Acid ◽  
Heterologous Expression ◽  
Lactobacillus Acidophilus ◽  
Lactobacillus Reuteri ◽  
Expression System ◽  
Fusion Partner ◽  
His Tag ◽  
Linoleic Acid Isomerase

The linoleic acid isomerase enzyme from Lactobacillus reuteri and Lactobacillus acidophilus, which were responsible for bioconversion of linoleic acid to cis-9, trans-11 conjugated linoleic acid (c9, t11 CLA) was cloned and overexpressed in Escherichia coli using expression system of pCold-SUMO. The gene was expressed under T7 promotor with a fusion partner of 6×His. Tag at its 5'terminal. After induction by 0.2mM IPTG for 18 h, the recombinant enzyme are expressing in the cytoplasm. The 1776 bp gene from L. reuteri and L. acidophilus encode a linoleic acid isomerase protein with 591 amino acids (64KD).

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