Effects of lytS-L on the primary metabolism and butenyl-spinosyn biosynthesis in Saccharopolyspora pogona

He, Haocheng; Peng, Shengnan; Yuan, Shuangqin; Tang, Jianli; Liu, Zhudong; Rang, Jie; Xia, Zi-Yuan; Hu, Jinjuan; Chen, Jianming; Ding, Xuezhi; Hu, Shengbiao; Sun, Yunjun; Xia, Liqiu

doi:10.1016/j.gene.2020.145130

Cited by 4 publications

(6 citation statements)

References 41 publications

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“…The MS parameters conditions were described in the previous study, and the proteins were searched and identified by SCIEX's ProteinPilot v 5.0. 24 Total RNA Isolation and qRT-PCR Analysis. To perform transcriptional analysis of genes, the biomass samples of different strains were harvested from the fermentation broth at different growth times.…”

Section: ■ Methodsmentioning

confidence: 99%

“…The collected solution was freeze-dried and dissolved in a 20 μL aqueous solution containing 0.1% formic acid and 2% acetonitrile, which was used for analysis by on-line nanoliter LC–MS. The MS parameters conditions were described in the previous study, and the proteins were searched and identified by SCIEX’s ProteinPilot v 5.0 …”

Section: Methodsmentioning

confidence: 99%

“…According to the differentially expressed proteins associated with the butenyl-spinosyn biosynthesis and growth of S. pogona (Table 3; Figure 5A), we found that busB, busC, busH, and busK appeared to be upregulated, which were directly synthesized by the butenylspinosyn. 24 To be specific, busB and busC are polyketide synthesis-related genes, and the synthesis of aglycone involves busH and busK.…”

Section: Deletion Of Clu13mentioning

confidence: 99%

“…The forosamine structure of butenyl-spinosyn is formed by BusO, BusN, BusQ, BusR, and BusS, and is connected to the aglycon under the catalysis of BusP. Furthermore, BusH, BusI, and BusK are responsible for the methylation modification of rhamnose. − To improve the yield of butenyl-spinosyn, a series of important proteins in S. pogona has been explored, such as polynucleotide phosphorylase (PNPase), LytT/LytR two-component system, Senx3/Regx3 two-component system, and TetR family regulators, − but these studies did not cause significant changes in butenyl-spinosyn production.…”

Section: Introductionmentioning

confidence: 99%

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Flaviolin-Like Gene Cluster Deletion Optimized the Butenyl-Spinosyn Biosynthesis Route in Saccharopolyspora pogona

Tang

Chen

et al. 2021

ACS Synth. Biol.

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Reduction and optimization of the microbial genome is an important strategy for constructing synthetic biological chassis cells and overcoming obstacles in natural product discovery and production. However, it is of great challenge to discover target genes that can be deleted and optimized due to the complicated genome of actinomycetes. Saccharopolyspora pogona can produce butenyl-spinosyn during aerobic fermentation, and its genome contains 32 different gene clusters. This suggests that there is a large amount of potential competitive metabolism in S. pogona, which affects the biosynthesis of butenyl-spinosyn. By analyzing the genome of S. pogona, six polyketide gene clusters were identified. From those, the complete deletion of clu13, a flaviolin-like gene cluster, generated a high butenyl-spinosyn-producing strain. Production of this strain was 4.06-fold higher than that of the wildtype strain. Transcriptome profiling revealed that butenyl-spinosyn biosynthesis was not primarily induced by the polyketide synthase RppA-like but was related to hypothetical protein Sp1764. However, the repression of sp1764 was not enough to explain the enormous enhancement of butenyl-spinosyn yields in S. pogona-Δclu13. After the comparative proteomic analysis of S. pogona-Δclu13 and S. pogona, two proteins, biotin carboxyl carrier protein (BccA) and response regulator (Reg), were investigated, whose overexpression led to great advantages of butenyl-spinosyn biosynthesis. In this way, we successfully discovered three key genes that obviously optimize the biosynthesis of butenyl-spinosyn. Gene cluster simplification performed in conjunction with multiomics analysis is of great practical significance for screening dominant chassis strains and optimizing secondary metabolism. This work provided an idea about screening key factors and efficient construction of production strains.

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Section: ■ Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Deletion Of Clu13mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Flaviolin-Like Gene Cluster Deletion Optimized the Butenyl-Spinosyn Biosynthesis Route in Saccharopolyspora pogona

Tang

Chen

et al. 2021

ACS Synth. Biol.

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“…pogona, and the production of butenyl-spinosyn increased by 4.72-fold compared to that in the original strain . Other genes, such as lytS-L , bfr , senX3-regX3 , sp1418 , sp_1288 , rhamnose synthetic genes, and metK , can also affect butenyl-spinosyn biosynthesis. − Although these reports demonstrate that some genes play important roles in butenyl-spinosyn biosynthesis, the genetic background of S. pogona, the metabolic network of butenyl-spinosyn biosynthesis, and the important functional genes involved in butenyl-spinosyn biosynthesis have not been revealed at the system level.…”

Section: Introductionmentioning

confidence: 97%

Promoting Butenyl-spinosyn Production Based on Omics Research and Metabolic Network Construction in Saccharopolyspora pogona

Rang

Cao

Shuai

et al. 2022

J. Agric. Food Chem.

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Understanding the metabolism of Saccharopolyspora pogona on a global scale is essential for manipulating its metabolic capabilities to improve butenyl-spinosyn biosynthesis. Here, we combined multiomics analysis to parse S. pogona genomic information, construct a metabolic network, and mine important functional genes that affect the butenyl-spinosyn biosynthesis. This research not only elucidated the relationship between butenyl-spinosyn biosynthesis and the primary metabolic pathway but also showed that the low expression level and continuous downregulation of the bus cluster and the competitive utilization of acetyl-CoA were the main reasons for reduced butenyl-spinosyn production. Our framework identified 148 genes related to butenyl-spinosyn biosynthesis that were significantly differentially expressed, confirming that butenyl-spinosyn polyketide synthase (PKS) and succinic semialdehyde dehydrogenase (GabD) play an important role in regulating butenyl-spinosyn biosynthesis. Combined modification of these genes increased overall butenyl-spinosyn production by 6.38-fold to 154.1 ± 10.98 mg/L. Our results provide an important strategy for further promoting the butenyl-spinosyn titer.

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Effects of salinity on the performance, microbial community, and functional genes among 4-chlorophenol wastewater treatment

Li,

Jiang,

Chen

et al. 2023

Bioresource Technology

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Effects of lytS-L on the primary metabolism and butenyl-spinosyn biosynthesis in Saccharopolyspora pogona

Cited by 4 publications

References 41 publications

Flaviolin-Like Gene Cluster Deletion Optimized the Butenyl-Spinosyn Biosynthesis Route in Saccharopolyspora pogona

Flaviolin-Like Gene Cluster Deletion Optimized the Butenyl-Spinosyn Biosynthesis Route in Saccharopolyspora pogona

Promoting Butenyl-spinosyn Production Based on Omics Research and Metabolic Network Construction in Saccharopolyspora pogona

Effects of salinity on the performance, microbial community, and functional genes among 4-chlorophenol wastewater treatment

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