Cloning and Heterologous Expression of Three Type II PKS Gene Clusters from <i>Streptomyces bottropensis</i>

Yan, Xiaohui; Probst, Katharina; Linnenbrink, Anton; Arnold, Moritz; Paululat, Thomas; Zeeck, Axel; Bechthold, Andreas

doi:10.1002/cbic.201100574

Cited by 33 publications

(41 citation statements)

References 61 publications

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“…The rishirilide biosynthetic gene cluster contains two luciferase-like monooxygenase genes [4]. Inactivation of rslO6 did not influence rishirilide production as described above.…”

Section: Galvaquinone a Is Produced By Mutants Lacking Rslo1mentioning

confidence: 66%

“…Rishirilide biosynthesis is controlled by four regulatory genes (rslR1, rslR2, rslR3 and rslR4) that belong to the family of SARP-regulatory genes (rslR1 and rslR2), the family of LAL regulatory genes (rslR3) and the family of MarR transcriptional regulatory genes (rslR4), respectively [4]. We observed that S. albus J1074 containing cos4 and pUWL-H-rslR1 + rslR2 + rslR3 (pUWLR1R2R3) produced more rishirilide B than S. albus J1074 containing only cos4.…”

Section: Overexpression Of the Regulatory Genes Rslr1 Rslr2 And Rslrmentioning

confidence: 99%

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Early Steps in the Biosynthetic Pathway of Rishirilide B

et al. 2020

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The biological active compound rishirilide B is produced by Streptomyces bottropensis. The cosmid cos4 contains the complete rishirilide B biosynthesis gene cluster. Its heterologous expression in the host Streptomyces albus J1074 led to the production of rishirilide B as a major compound and to small amounts of rishirilide A, rishirilide D and lupinacidin A. In order to gain more insights into the biosynthesis, gene inactivation experiments and gene expression experiments were carried out. This study lays the focus on the functional elucidation of the genes involved in the early biosynthetic pathway. A total of eight genes were deleted and six gene cassettes were generated. Rishirilide production was not strongly affected by mutations in rslO2, rslO6 and rslH. The deletion of rslK4 and rslO3 led to the formation of polyketides with novel structures. These results indicated that RslK4 and RslO3 are involved in the generation or selection of the starter unit for rishirilide biosynthesis. In the rslO10 mutant strain, two novel compounds were detected, which were also produced by a strain containing solely the genes rslK1, rslK2, rslK3, rslK4, and rslA. rslO1 and rslO4 mutants predominately produce galvaquinones. Therefore, the ketoreductase RslO10 is involved in an early step of rishirilide biosynthesis and the oxygenases RslO1 and RslO4 are most probably acting on an anthracene moiety. This study led to the functional elucidation of several genes of the rishirilide pathway, including rslK4, which is involved in selecting the unusual starter unit for polyketide synthesis.Molecules 2020, 25, 1955 2 of 15 the initiation module, can be found, consisting of a ketosynthase, a malonyl CoA:ACP transferase and an acyl carrier protein (ACP). The initiation module selects the starter unit and catalyzes the first elongation of the growing polyketide, before the molecule is loaded onto the minPKS for further elongation reactions. Additional enzymes, like ketoreductases, cyclases and aromatases complement the minPKS and determine the folding of the respective polyketide [1]. The aromatic polyketide rishirilide B (Figure 1) was first discovered by Iwaki et al. in 1984. Rishirilide B was isolated from Streptomyces rishiriensis OFR-1056 in the course of screening for new α2-macroglobulin inhibitors [2]. This compound was also described as a product of S. olivaceus SCSIO T05, which also produced rishirilide C, lupinacidin A and galvaquinone A and B [3] (Figure 1). The biosynthetic gene cluster of this strain responsible for the formation of all four compounds was cloned and biosynthetic studies were performed [3]. S. bottropensis (formerly S. sp. Gö C4/4) also produces the compounds rishirilide A, B, D and lupinacidin A (Figure 1). The corresponding rishirilide biosynthetic gene cluster was cloned on a cosmid (cos4), which was subsequently expressed in the heterologous host S. albus J1074. Rishirilide B was produced as a major compound. Moreover, rishirilide A, D and lupinacidin A were also produced in small amounts [3] (Figure 1)....

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“…The rishirilide biosynthetic gene cluster contains two luciferase-like monooxygenase genes [4]. Inactivation of rslO6 did not influence rishirilide production as described above.…”

Section: Galvaquinone a Is Produced By Mutants Lacking Rslo1mentioning

confidence: 66%

Section: Overexpression Of the Regulatory Genes Rslr1 Rslr2 And Rslrmentioning

confidence: 99%

Early Steps in the Biosynthetic Pathway of Rishirilide B

et al. 2020

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“…GTRI‐02 is also produced by Streptomyces sp. strains GW4184, ANK313, and Gö C4/4, and recently was identified in Streptomyces violaceoruber and Streptomyces coelicolor A3(2) . According to a bio‐retrosynthetic analysis (using fungal biogenesis), 2 is synthesized chemoenzymatically by use of a fungal enzyme.…”

Section: Methodsmentioning

confidence: 99%

Insights into the Role of Ketoreductases in the Biosynthesis of Partially Reduced Bacterial Aromatic Polyketides*

et al. 2019

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Partially reduced aromatic polyketides are bioactive secondary metabolites or intermediates in the biosynthesis of deoxygenated aromatics. For the antibiotic GTRI‐02 (mensalone) in different Streptomyces spp., biosynthesis involving the reduction of a fully aromatized acetyltrihydroxynaphthalene by a naphthol reductase has been proposed and shown in vitro with a fungal enzyme. However, more recently, GTRI‐02 has been identified as a product of the ActIII biosynthetic gene cluster from Streptomyces coelicolor A3(2), for which the reduction of a linear polyketide precursor by ActIII ketoreductase, prior to cyclization and aromatization, has been suggested. We have examined three different ketoreductases from bacterial producer strains of GTRI‐02 for their ability to reduce mono‐, bi‐, and tricyclic aromatic substrates. The enzymes reduced 1‐ and 2‐tetralone but not other aromatic substrates. This strongly suggests a reduction of a cyclized but not yet aromatic polyketide intermediate in the biosynthesis of GTRI‐02. Implications of the results for the biosynthesis of other secondary polyketidic metabolites are discussed.

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“…[7,8] Therefore,w e compared the spectral and rotation data, and HPLC retention time,o fo ur synthetic (2R,3R,4R)-2 with those of natural (+ +)-2,w hich was kindly provided by Professor Dr.B echthold. [2] The 1 HNMR spectrum of our synthetic (2R,3R,4R)-2 matched that of natural (+ +)-2,but the optical rotation of the natural product had aplus sign [ a ½ 25 D ¼ + 46.7 (c 0.03, EtOH)], and the HPLC retention time of natural (+ +)-2 (t = 8.3 min) was different from that of our synthetic (2R,3R,4R)-2 (t = 11.2 min) on achiral-phase column (Figure 2a…”

Section: Communicationsmentioning

confidence: 99%

“…(À)-Rishirilide A( 1)a nd (+ +)-rishirilide B[ ( + +)-2]a re anthracenone-type natural products isolated from Streptomyces rishiriensis OFR-1056 in 1984 ( Figure 1), [1] and were subsequently found in Streptomyces bottropensis. [2] (+ +)-Rishirilide B[(+ +)-2]has moderate inhibitory activities against a 2macroglobulin, glutathione S-transferase,a nd asparaginyl-tRNAs ynthetase, [3,4] which play roles in thrombosis and cancer. Key structural features of the rishirilides include three or four consecutive stereogenic centers involving quaternary carbon centers on the Aring.…”

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confidence: 99%

Total Synthesis of Rishirilide B by Organocatalytic Oxidative Kinetic Resolution: Revision of Absolute Configuration of (+)‐Rishirilide B

et al. 2017

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Described herein is the enantioselective syntheses of (+)- and (-)-rishirilide B from the corresponding optically active β-substituted tetralones, which were obtained by oxidative kinetic resolution based on α-hydroxylation in the presence of a chiral guanidine-bisurea bifunctional organocatalyst. Benzylic oxidation of the tetralones at C1 followed by regioselective isomerization of the oxabenzonorbornadiene structure led to rishirilide B. Our findings lead to the revision of the previously proposed (2R,3R,4R) absolute configuration of (+)-rishirilide B to (2S,3S,4S).

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Cloning and Heterologous Expression of Three Type II PKS Gene Clusters from Streptomyces bottropensis

Cited by 33 publications

References 61 publications

Early Steps in the Biosynthetic Pathway of Rishirilide B

Early Steps in the Biosynthetic Pathway of Rishirilide B

Insights into the Role of Ketoreductases in the Biosynthesis of Partially Reduced Bacterial Aromatic Polyketides*

Total Synthesis of Rishirilide B by Organocatalytic Oxidative Kinetic Resolution: Revision of Absolute Configuration of (+)‐Rishirilide B

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