Biosynthesis of plant-specific phenylpropanoids by construction of an artificial biosynthetic pathway in Escherichia coli.

Choi, Oksik,Wu, Cheng-Zhu,Kang, Sun Young,Ahn, Jong Seog,Uhm, Tai-Boong,Hong, Young-Soo Published by Stockton Press on behalf of the Socie 2011 Journal of industrial microbiology & biotechnology Vol.38 No.10

<P>Biological synthesis of plant secondary metabolites has attracted increasing attention due to their proven or assumed beneficial properties and health-promoting effects. Phenylpropanoids are the precursors to a range of important plant metabolites such as the secondary metabolites belonging to the flavonoid/stilbenoid class of compounds. In this study, engineered Escherichia coli containing artificial phenylpropanoid biosynthetic pathways utilizing tyrosine as the initial precursor were established for production of plant-specific metabolites such as ferulic acid, naringenin, and resveratrol. The construction of the artificial pathway utilized tyrosine ammonia lyase and 4-coumarate 3-hydroxylase from Saccharothrix espanaensis, cinnamate/4-coumarate:coenzyme A ligase from Streptomyces coelicolor, caffeic acid O-methyltransferase and chalcone synthase from Arabidopsis thaliana, and stilbene synthase from Arachis hypogaea.</P>

Construction of Artificial Biosynthetic Pathways for Resveratrol Glucoside Derivatives(s)

( Oksik Choi ),( Jae Kyoung Lee ),( Sun Young Kang ),( Ramesh Prasad Pandey ),( Jae Kyung Sohng ),( Jong Seog Ahn ),( Young Soo Hong ) 한국미생물 · 생명공학회 2014 Journal of microbiology and biotechnology Vol.24 No.5

Resveratrol, which is a polyphenolic antioxidant, is dose-dependent when used to provide health benefits, to enhance stress resistance, and to extend lifespans. However, even though resveratrol has therapeutic benefits, its clinical therapeutic effect is limited owing to its low oral bioavailability. An Escherichia coli system was developed that contains an artificial biosynthetic pathway that produces resveratrol glucoside derivatives, such as resveratrol-3-Oglucoside (piceid) and resveratrol-4`-O-glucoside (resveratroloside), from simple carbon sources. This artificial biosynthetic pathway contains a glycosyltransferase addition (YjiC from Bacillus) with resveratrol biosynthetic genes. The produced glucoside compounds were verified through the presence of a product peak(s) and also through LC/MS analyses. The strategy used in this research demonstrates the first harnessing of E. coli for de novo synthesis of resveratrol glucoside derivatives from a simple sugar medium.

6-Alkylsalicylic Acid Analogues Inhibit In Vitro ATPase Activity of Heat Shock Protein 90

Cheng-Zhu Wu,An Na Moon,Oksik Choi,Sun-Young Kang,Jung Joon Lee,황방연,김영호,Hong-Sub Lee,홍영수,이동호 대한약학회 2010 Archives of Pharmacal Research Vol.33 No.12

The molecular chaperone heat shock protein 90 (Hsp90) is responsible for maintaining the correct folding and stability of many signaling proteins. It is a promising target of cancer therapeutics and several other diseases, including neurodegenerative disease, nerve injuries, inflammation, and infection. In an effort to identify new Hsp90 inhibitors from natural sources using an in vitro ATPase inhibition assay, two 6-alkylsalicylic acid analogues, salaceyin A and B were identified from the culture extract of Streptomyces. Salaceyin A and B exhibited moderate ATPase inhibitory activities with IC50 values of 68.3 and 65.2 μM, respectively. Binding of salaceyins to human Hsp90α was examined by competition binding experiments with ATPSepharose beads. However, the compounds exhibited no degradation activity of Hsp90 client proteins, Her2, c-Raf, or Akt.

Colorectal Cancer Therapy Using a Pediococcus pentosaceus SL4 Drug Delivery System Secreting Lactic Acid Bacteria-Derived Protein p8

정명준,안병철,Yongku Ryu,Yeo-Sang Yoon,Oksik Choi,박호진,김태엽,김송인,김봉규 한국분자세포생물학회 2019 Molecules and cells Vol.42 No.11

Despite decades of research into colorectal cancer (CRC), there is an ongoing need for treatments that are more effective and safer than those currently available. Lactic acid bacteria (LAB) show beneficial effects in the context of several diseases, including CRC, and are generally regarded as safe. Here, we isolated a Lactobacillus rhamnosus (LR)-derived therapeutic protein, p8, which suppressed CRC proliferation. We found that p8 translocated specifically to the cytosol of DLD-1 cells. Moreover, p8 down-regulated expression of Cyclin B1 and Cdk1, both of which are required for cell cycle progression. We confirmed that p8 exerted strong anti-proliferative activity in a mouse CRC xenograft model. Intraperitoneal injection of recombinant p8 (r-p8) led to a significant reduction (up to 59%) in tumor mass when compared with controls. In recent years, bacterial drug delivery systems (DDSs) have proven to be effective therapeutic agents for acute colitis. Therefore, we aimed to use such systems, particularly LAB, to generate the valuable therapeutic proteins to treat CRC. To this end, we developed a gene expression cassette capable of inducing secretion of large amounts of p8 protein from Pediococcus pentosaceus SL4 (PP). We then confirmed that this protein (PP-p8) exerted anti-proliferative activity in a mouse CRC xenograft model. Oral administration of PP-p8 DDS led to a marked reduction in tumor mass (up to 64%) compared with controls. The PP-p8 DDS using LAB described herein has advantages over other therapeutics; these advantages include improved safety (the protein is a probiotic), cost-free purification, and specific targeting of CRC cells.

Colorectal Cancer Therapy Using a Pediococcus pentosaceus SL4 Drug Delivery System Secreting Lactic Acid Bacteria-Derived Protein p8

An, Byung Chull,Ryu, Yongku,Yoon, Yeo-Sang,Choi, Oksik,Park, Ho Jin,Kim, Tai Yeub,Kim, Song-In,Kim, Bong-Kyu,Chung, Myung Jun Korean Society for Molecular and Cellular Biology 2019 Molecules and cells Vol.42 No.11

Despite decades of research into colorectal cancer (CRC), there is an ongoing need for treatments that are more effective and safer than those currently available. Lactic acid bacteria (LAB) show beneficial effects in the context of several diseases, including CRC, and are generally regarded as safe. Here, we isolated a Lactobacillus rhamnosus (LR)-derived therapeutic protein, p8, which suppressed CRC proliferation. We found that p8 translocated specifically to the cytosol of DLD-1 cells. Moreover, p8 down-regulated expression of Cyclin B1 and Cdk1, both of which are required for cell cycle progression. We confirmed that p8 exerted strong anti-proliferative activity in a mouse CRC xenograft model. Intraperitoneal injection of recombinant p8 (r-p8) led to a significant reduction (up to 59%) in tumor mass when compared with controls. In recent years, bacterial drug delivery systems (DDSs) have proven to be effective therapeutic agents for acute colitis. Therefore, we aimed to use such systems, particularly LAB, to generate the valuable therapeutic proteins to treat CRC. To this end, we developed a gene expression cassette capable of inducing secretion of large amounts of p8 protein from Pediococcus pentosaceus SL4 (PP). We then confirmed that this protein (PP-p8) exerted anti-proliferative activity in a mouse CRC xenograft model. Oral administration of PP-p8 DDS led to a marked reduction in tumor mass (up to 64%) compared with controls. The PP-p8 DDS using LAB described herein has advantages over other therapeutics; these advantages include improved safety (the protein is a probiotic), cost-free purification, and specific targeting of CRC cells.

Rational Biosynthetic Engineering for Optimization of Geldanamycin Analogues

Kim, Woncheol,Lee, Dongho,Hong, Seong Su,Na, Zhu,Shin, Jin Chul,Roh, Su Heun,Wu, Cheng-Zhu,Choi, Oksik,Lee, Kyeong,Shen, Yue-Mao,Paik, Sang-Gi,Lee, Jung Joon,Hong, Young-Soo WILEY-VCH Verlag 2009 Chembiochem Vol.10 No.7

<P>Tailor made: We report the rational biosynthesis of C15 hydroxylated non-quinone geldanamycin analogues by site-directed mutagenesis of the geldanamycin polyketide synthase (PKS), together with a combination of post-PKS tailoring genes. Rational biosynthetic engineering allowed the generation of geldanamycin derivatives, such as DHQ3 illustrated in the figure, which had superior pharmacological properties in comparison to the parent compound. <img src='wiley_img/14394227-2009-10-7-CBIC200800763-content.gif' alt='wiley_img/14394227-2009-10-7-CBIC200800763-content'> </P><P>A rational biosynthetic engineering approach was applied to the optimization of the pharmacological properties of the benzoquinone ansamycin, geldanamycin. Geldanamycin and its natural or semisynthetic derivatives have the potential to serve as anticancer chemotherapeutic agents. However, these first-generation Hsp90 inhibitors share an unfavorable structural feature that causes both reduced efficacy and toxicity during clinical evaluation. We report the rationally designed biosynthesis of C15 hydroxylated non-quinone geldanamycin analogues by site-directed mutagenesis of the geldanamycin polyketide synthase (PKS), together with a combination of post-PKS tailoring genes. A 15-hydroxyl-17-demethoxy non-quinone analogue, DHQ3, exhibited stronger inhibition of Hsp90 ATPase activity (4.6-fold) than geldanamycin. Taken together, the results of the present study indicate that rational biosynthetic engineering allows the generation of derivatives of geldanamycin with superior pharmacological properties.</P> <B>Graphic Abstract</B> <P>Tailor made: We report the rational biosynthesis of C15 hydroxylated non-quinone geldanamycin analogues by site-directed mutagenesis of the geldanamycin polyketide synthase (PKS), together with a combination of post-PKS tailoring genes. Rational biosynthetic engineering allowed the generation of geldanamycin derivatives, such as DHQ3 illustrated in the figure, which had superior pharmacological properties in comparison to the parent compound. <img src='wiley_img/14394227-2009-10-7-CBIC200800763-content.gif' alt='wiley_img/14394227-2009-10-7-CBIC200800763-content'> </P>