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Farnesol production from <i>Escherichia coli</i> by harnessing the exogenous mevalonate pathway
Wang, Chonglong,Yoon, Sang‐,Hwal,Shah, Asad Ali,Chung, Young‐,Ryun,Kim, Jae‐,Yean,Choi, Eui‐,Sung,Keasling, Jay D.,Kim, Seon‐,Won Wiley Subscription Services, Inc., A Wiley Company 2010 Biotechnology and bioengineering Vol.107 No.3
<P><B>Abstract</B></P><P>Farnesol (FOH) production has been carried out in metabolically engineered <I>Escherichia coli</I>. FOH is formed through the depyrophosphorylation of farnesyl pyrophosphate (FPP), which is synthesized from isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP) by FPP synthase. In order to increase FPP synthesis, <I>E. coli</I> was metabolically engineered to overexpress <I>ispA</I> and to utilize the foreign mevalonate (MVA) pathway for the efficient synthesis of IPP and DMAPP. Two‐phase culture using a decane overlay of the culture broth was applied to reduce volatile loss of FOH produced during culture and to extract FOH from the culture broth. A FOH production of 135.5 mg/L was obtained from the recombinant <I>E. coli</I> harboring the pTispA and pSNA plasmids for <I>ispA</I> overexpression and MVA pathway utilization, respectively. It is interesting to observe that a large amount of FOH could be produced from <I>E. coli</I> without FOH synthase by the augmentation of FPP synthesis. Introduction of the exogenous MVA pathway enabled the dramatic production of FOH by <I>E. coli</I> while no detectable FOH production was observed in the endogenous MEP pathway‐only control. Biotechnol. Bioeng. 2010;107: 421–429. © 2010 Wiley Periodicals, Inc.</P>
Reassessing <i>Escherichia coli</i> as a cell factory for biofuel production
Wang, Chonglong,Pfleger, Brian F,Kim, Seon-Won Elsevier 2017 Current opinion in biotechnology Vol.45 No.-
<P>Via metabolic engineering, industrial microorganisms have the potential to convert renewable substrates into a wide range of biofuels that can address energy security and environmental challenges associated with current fossil fuels. The user-friendly bacterium, <I>Escherichia coli</I>, remains one of the most frequently used hosts for demonstrating production of biofuel candidates including alcohol-, fatty acid- and terpenoid-based biofuels. In this review, we summarize the metabolic pathways for synthesis of these biofuels and assess enabling technologies that assist in regulating biofuel synthesis pathways and rapidly assembling novel <I>E. coli</I> strains. These advances maintain <I>E. coli</I>’s position as a prominent host for developing cell factories for biofuel production.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Biofuel production has been achieved in <I>Escherichia coli</I> by construction of several metabolic pathways. </LI> <LI> The emerging technologies shows great potential in pathway engineering and stain manipulation. </LI> <LI> Tolerance engineering was required to construct an ideal biofuel producing <I>E. coli</I> host. </LI> </UL> </P>
Wang, Chonglong,Zada, Bakht,Wei, Gongyuan,Kim, Seon-Won Elsevier 2017 Bioresource technology Vol.241 No.-
<P><B>Abstract</B></P> <P>Isoprenoids comprise the largest family of natural organic compounds with many useful applications in the pharmaceutical, nutraceutical, and industrial fields. Rapid developments in metabolic engineering and synthetic biology have facilitated the engineering of isoprenoid biosynthetic pathways in <I>Escherichia coli</I> to induce high levels of production of many different isoprenoids. In this review, the stem pathways for synthesizing isoprene units as well as the branch pathways deriving diverse isoprenoids from the isoprene units have been summarized. The review also highlights the metabolic engineering efforts made for the biosynthesis of hemiterpenoids, monoterpenoids, sesquiterpenoids, diterpenoids, carotenoids, retinoids, and coenzyme Q<SUB>10</SUB> in <I>E</I>. <I>coli</I>. Perspectives and future directions for the synthesis of novel isoprenoids, decoration of isoprenoids using cytochrome P450 enzymes, and secretion or storage of isoprenoids in <I>E</I>. <I>coli</I> have also been included.</P> <P><B>Highlights</B></P> <P> <UL> <LI> This review covered production of isoprenoid in <I>E</I>. <I>coli</I> over decades. </LI> <LI> This review summarized progresses of pathway engineering for isoprenoid production. </LI> <LI> This review suggested three directions for isoprenoid production in the future. </LI> </UL> </P>
Wang, Chonglong,Park, Ju‐,Eon,Choi, Eui‐,Sung,Kim, Seon‐,Won WILEY‐VCH Verlag 2016 BIOTECHNOLOGY JOURNAL Vol.11 No.10
<P><B>Abstract</B></P><P>Farnesol is a sesquiterpenoid alcohol that has important industrial and medical potential. It is usually synthesized from farnesyl diphosphate (FPP) by farnesol synthase in plants. FPP accumulation can cause up‐regulation of phosphatases capable of FPP hydrolysis, resulting in farnesol production in <I>Escherichia coli</I>. We found that PgpB and YbjG, two integral membrane phosphatases, can hydrolyze FPP into farnesol. Overexpression of FPP synthase (IspA) and PgpB, along with a heterologous mevalonate pathway, enabled recombinant <I>E. coli</I> to produce 526.1 mg/L of farnesol. This result indicates that the phosphatases PgpB and YbjG can be used to construct a novel farnesol synthesis pathway for mass production in <I>E. coli</I>.</P>
Jingen Xu,Chonglong Wang,Erhui Jin,Youfang Gu,Shenghe Li,Qinggang Li 한국유전학회 2018 Genes & Genomics Vol.40 No.4
Intramuscular fat (IMF) content is an important trait closely related to meat quality, which is highly variable among pig breeds from diverse genetic backgrounds. High-throughput sequencing has become a powerful technique for analyzing the whole transcription profiles of organisms. In order to elucidate the molecular mechanism underlying porcine meat quality, we adopted RNA sequencing to detect transcriptome in the longissimus dorsi muscle of Wei pigs (a Chinese indigenous breed) and Yorkshire pigs (a Western lean-type breed) with different IMF content. For the Wei and Yorkshire pig libraries, over 57 and 64 million clean reads were generated by transcriptome sequencing, respectively. A total of 717 differentially expressed genes (DEGs) were identified in our study (false discovery rate < 0.05 and fold change > 2), with 323 up-regulated and 394 down-regulated genes in Wei pigs compared with Yorkshire pigs. Gene Ontology analysis showed that DEGs significantly related to skeletal muscle cell differentiation, phospholipid catabolic process, and extracellular matrix structural constituent. Pathway analysis revealed that DEGs were involved in fatty acid metabolism, steroid biosynthesis, glycerophospholipid metabolism, and protein digestion and absorption. Quantitative real time PCR confirmed the differential expression of 11 selected DEGs in both pig breeds. The results provide useful information to investigate the transcriptional profiling in skeletal muscle of different pig breeds with divergent phenotypes, and several DEGs can be taken as functional candidate genes related to lipid metabolism (ACSL1, FABP3, UCP3 and PDK4) and skeletal muscle development (ASB2, MSTN, ANKRD1 and ANKRD2).