Insights into Bacterial 6-Methylsalicylic Acid Synthase and Its Engineering to Orsellinic Acid Synthase for Spirotetronate Generation Wei Ding, Chun.

Slides:

Advertisements

Similar presentations

Volume 13, Issue 6, Pages (June 2006)

Advertisements

Volume 22, Issue 5, Pages v-vi (May 2015)

Volume 19, Issue 9, Pages (September 2012)

Volume 16, Issue 10, Pages (October 2009)

Foundations for Directed Alkaloid Biosynthesis

Volume 15, Issue 7, Pages (July 2008)

Volume 20, Issue 10, Pages (October 2013)

Volume 15, Issue 2, Pages (February 2008)

Adding Specificity to Artificial Transcription Activators

Volume 17, Issue 4, Pages (April 2010)

Volume 17, Issue 4, Pages (April 2010)

Volume 13, Issue 4, Pages (April 2006)

Volume 17, Issue 2, Pages (February 2010)

Volume 9, Issue 3, Pages (March 2002)

Biosynthesis of Actinorhodin and Related Antibiotics: Discovery of Alternative Routes for Quinone Formation Encoded in the act Gene Cluster Susumu Okamoto,

Benjamin N. Mijts, Pyung Cheon Lee, Claudia Schmidt-Dannert

Volume 19, Issue 2, Pages (February 2012)

Volume 14, Issue 5, Pages (May 2007)

No Need To Be Pure: Mix the Cultures!

Volume 17, Issue 10, Pages (October 2010)

Elucidation of the Biosynthetic Gene Cluster and the Post-PKS Modification Mechanism for Fostriecin in Streptomyces pulveraceus Rixiang Kong, Xuejiao.

Volume 19, Issue 3, Pages (March 2012)

Volume 14, Issue 2, Pages (February 2007)

Volume 12, Issue 12, Pages (December 2005)

Structure-Based Dissociation of a Type I Polyketide Synthase Module

Volume 18, Issue 4, Pages (April 2011)

PqsE of Pseudomonas aeruginosa Acts as Pathway-Specific Thioesterase in the Biosynthesis of Alkylquinolone Signaling Molecules Steffen Lorenz Drees,

Volume 20, Issue 4, Pages (April 2013)

Volume 15, Issue 11, Pages (November 2008)

Volume 10, Issue 11, Pages (November 2003)

Volume 17, Issue 1, Pages (January 2010)

Volume 16, Issue 5, Pages (May 2009)

Volume 17, Issue 7, Pages (July 2010)

Volume 21, Issue 8, Pages v-vi (August 2014)

Formation of functional heterologous complexes using subunits from the picromycin, erythromycin and oleandomycin polyketide synthases Li Tang, Hong Fu,

Volume 15, Issue 2, Pages (February 2008)

Foundations for Directed Alkaloid Biosynthesis

Volume 20, Issue 6, Pages (June 2013)

In Vivo Characterization of Nonribosomal Peptide Synthetases NocA and NocB in the Biosynthesis of Nocardicin A Jeanne M. Davidsen, Craig A. Townsend

Volume 11, Issue 10, Pages (October 2004)

Volume 17, Issue 4, Pages (April 2010)

Volume 15, Issue 8, Pages (August 2008)

Volume 15, Issue 6, Pages (June 2008)

Volume 23, Issue 2, Pages (February 2016)

Engineered Biosynthesis of Geldanamycin Analogs for Hsp90 Inhibition

Volume 16, Issue 5, Pages (May 2009)

Characterization of the Biosynthetic Gene Cluster for Benzoxazole Antibiotics A33853 Reveals Unusual Assembly Logic Meinan Lv, Junfeng Zhao, Zixin Deng,

Volume 13, Issue 3, Pages (March 2006)

Gerald Lackner, Markus Bohnert, Jonas Wick, Dirk Hoffmeister

Volume 11, Issue 3, Pages (March 2004)

Volume 11, Issue 12, Pages (December 2004)

Volume 16, Issue 2, Pages (February 2009)

Volume 12, Issue 2, Pages (February 2005)

Leinamycin Biosynthesis Revealing Unprecedented Architectural Complexity for a Hybrid Polyketide Synthase and Nonribosomal Peptide Synthetase Gong-Li.

Biosynthetic Pathway Connects Cryptic Ribosomally Synthesized Posttranslationally Modified Peptide Genes with Pyrroloquinoline Alkaloids Peter A. Jordan,

L-DOPA Ropes in tRNAPhe

Aza-Tryptamine Substrates in Monoterpene Indole Alkaloid Biosynthesis

Volume 22, Issue 8, Pages (August 2015)

Dual Carbamoylations on the Polyketide and Glycosyl Moiety by Asm21 Result in Extended Ansamitocin Biosynthesis Yan Li, Peiji Zhao, Qianjin Kang, Juan.

Volume 21, Issue 9, Pages (September 2014)

Volume 18, Issue 8, Pages (August 2011)

Cloning, Heterologous Expression, and Characterization of the Gene Cluster Required for Gougerotin Biosynthesis Guoqing Niu, Lei Li, Junhong Wei, Huarong.

Volume 15, Issue 7, Pages (July 2008)

Biomimetic Thioesters as Probes for Enzymatic Assembly Lines: Synthesis, Applications, and Challenges Jakob Franke, Christian Hertweck Cell Chemical.

Volume 12, Issue 3, Pages (March 2005)

Volume 12, Issue 3, Pages (March 2005)

Volume 13, Issue 3, Pages (March 2006)

Volume 21, Issue 9, Pages (September 2014)

Volume 12, Issue 10, Pages (October 2005)

Volume 22, Issue 4, Pages vii-viii (April 2015)

Presentation transcript:

Insights into Bacterial 6-Methylsalicylic Acid Synthase and Its Engineering to Orsellinic Acid Synthase for Spirotetronate Generation Wei Ding, Chun Lei, Qingli He, Qinglin Zhang, Yurong Bi, Wen Liu Chemistry & Biology Volume 17, Issue 5, Pages 495-503 (May 2010) DOI: 10.1016/j.chembiol.2010.04.009 Copyright © 2010 Elsevier Ltd Terms and Conditions

Figure 1 6-Methylsalicylic Acid Synthase and Orsellinic Acid Synthase, and Biosyntheses of 6-MSA, Triacetic Acid Lactone, and Orsellinic Acid (A) Domain organizations of 6-methylsalicylic acid synthase (6-MSAS) and orsellinic acid synthase (OSAS). KS, ketoacylsynthase; AT, acyltransferase; DH, dehydratase; KR, ketoreductase; ACP, acyl carrier protein. (B) Pathways for 6-MSA synthesized by 6-MSASs and DH mutant ChlB1 (H947F), for triacetic acid lactone (TAL) synthesized by KR-mutated fungal 6-MSASs (AxPxxA at the NADPH binding site), and for orsellinic acid (OSA) synthesized by OSASs AviM, CalO5, and KR-mutated ChlB1 (Y1450F at the active site). For 6-MSASs, solid arrows indicate native enzyme-catalyzed reactions, and dashed arrows showed the shunt reactions catalyzed by mutant enzymes. Chemistry & Biology 2010 17, 495-503DOI: (10.1016/j.chembiol.2010.04.009) Copyright © 2010 Elsevier Ltd Terms and Conditions

Figure 2 Functional Validation of DH or KR Mutant ChlB1 Derivatives in S. albus HPLC analysis of the products from the negative control TL1006 (carrying the vector pTGV-2) (I); standard 6-MSA (II, asterisk); TL1071 for the native ChlB1 (III); TL1072 for the DH mutant with H947A (IV); TL1073 for the DH mutant with H947F (V); standard TAL (VI, dot); standard OSA (VII, triangle); TL1074 for the KR mutant with G1389A (VIII); TL1075 for the KR mutant with G1387A, G1389P, and G1392A (XI); and TL1076 for the KR mutant with Y1540F (X). Chemistry & Biology 2010 17, 495-503DOI: (10.1016/j.chembiol.2010.04.009) Copyright © 2010 Elsevier Ltd Terms and Conditions

Figure 3 Aryl Group Formation and Incorporation in CHL Biosynthesis ChlB1 or its KR mutant (Y1540F) postmodifications of the 6-MSA or OSA moiety and its transfer to DM-CHL for affording deschloro-CHL and CHL or the corresponding analogs 7 and 8. The dashed circle indicates the variable aryl group. Chemistry & Biology 2010 17, 495-503DOI: (10.1016/j.chembiol.2010.04.009) Copyright © 2010 Elsevier Ltd Terms and Conditions

Figure 4 Validation of New Spirotetronate Production in S. antibioticus HPLC analysis of fermentation cultures from the wild-type strain (I), chlB1-inactivated mutant TL1012 (II), and TL1077 by in trans complementation of the construct for expressing the KR mutant ChlB1 (Y1540F) in TL1012 (III). a, deschloro-CHL; b, CHL; c, DM-CHL; d, compound 7; and e, compound 8. Chemistry & Biology 2010 17, 495-503DOI: (10.1016/j.chembiol.2010.04.009) Copyright © 2010 Elsevier Ltd Terms and Conditions

Figure 5 Structural Elucidation of New Spirotetronates 1H-1H COSY (for 7) and selected HMBC correlations (for 7 and 8) that establish the variable substitutions on the aryl group of new spirotetronates. Chemistry & Biology 2010 17, 495-503DOI: (10.1016/j.chembiol.2010.04.009) Copyright © 2010 Elsevier Ltd Terms and Conditions