Dataset Information

Proteomic analyses reveal metabolic responses to 3-hydroxypropionic acid synthesized internally in Synechocystis sp. PCC 6803

ABSTRACT: Using an engineered 3-HP producing SM strain, in this project, the cellular responses to 3-HP internally produced were first determined using a quantitative iTRAQ-LC-MS/MS proteomics approach. A total of 2,264 unique proteins were identified, which represented about 63% of all predicted protein in Synechocystis in the proteomic analysis of the 3-HP producing strain SM and wild type. Among all identified proteins, 204 proteins were found up- regulated and 123 proteins were found down-regulated, respectively. The proteins related to oxidative phosphorylation, photosynthesis, ribosome, central carbon metabolism, two-component systems and ABC-type transporters were up-regulated. The results suggested that the supply of ATP and NADPH was increased significantly, and the precursor malonyl-CoA and acetyl-CoA may also be supplemented when 3-HP was produced at a high level in Synechocystis.

INSTRUMENT(S): 4700 Proteomics Analyzer

ORGANISM(S): Synechococcus Cyanophage Syn

SUBMITTER: Yunpeng Wang

LAB HEAD: Weiwen Zhang

PROVIDER: PXD004974 | Pride | 2016-09-26

REPOSITORIES: Pride

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Publications

Proteomic and metabolomic analyses reveal metabolic responses to 3-hydroxypropionic acid synthesized internally in cyanobacterium <i>Synechocystis</i> sp. PCC 6803.

Wang Yunpeng Y Chen Lei L Zhang Weiwen W

Biotechnology for biofuels 20161006

<h4>Background</h4>3-hydroxypropionic acid (3-HP) is an important platform chemical with a wide range of applications. In our previous study, the biosynthetic pathway of 3-HP was constructed and optimized in cyanobacterium <i>Synechocystis</i> sp. PCC 6803, which led to 3-HP production directly from CO<sub>2</sub> at a level of 837.18 mg L<sup>-1</sup> (348.8 mg/g dry cell weight). As the production and accumulation of 3-HP in cells affect cellular metabolism, a better understanding of cellular ...[more]

PMID: 27757169

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Project description:In the present study, we have used a genomic approach to understand how the classical mutate-and-screen method actually generated an improved rifamycin B producer. Compared with the reference strains Amycolatopsis mediterranei S699 (rifamycin B producer) and U32 (rifamycin SV producer), a total of 250 variations affecting a total of 227 coding sequences (CDS) were found in rifamycin B overproducing strain HP-130. One hundred nine CDS variations were specific to HP-130 as they were absent in both S699 and U32. A lot of variations affected genes coding for fatty acid (an lipid) metabolism, which is tightly interconnected with rifamycin biosynthesis by common metabolic precursors (malonyl-CoA, methylmalonyl-CoA). Interestingly, a nonsense mutation was mapped within mutB coding for methylmalonyl-CoA mutase suggesting the importance of metabolic re-direction of carbon flow toward rifamycin biosynthesis in the over-producing phenotype of HP-130. Other key mutations were: i.) a missense mutation affecting ppk, which encodes a polyphosphate kinase and was previously shown to play a negative role in the control of antibiotic biosynthesis in Streptomyces lividans; ii.) a missense mutation in argS2 affecting one of the two arginyl tRNA synthetases of A. mediterranei; iii.) a missense mutation in rifN coding for a protein belonging to the ROK family of transcriptional regulators with kanosamine kinase activity. Microarray analysis of the transcriptome of HP-130 as compared to that of S699 was useful to understand the effects of several mutations on global gene expression profile. Genomic and transcriptomic data were used to improve rifamycin B production in S699 by genetic engineering thus proving the causative effect of the above-mentioned mutations on the overproducing phenotype. In this study by using comparative genomic analysis we have identified all genetic changes that have occurred during development of a rifamycin B overproducer (HP-130), which was obtained by the traditional mutate-and-screen method. To gain insight about the mechanisms underlying improved rifamycin B production in HP-130, DNA microarray of A. mediterranei were manufactured and used for comparative analysis of the gene expression profile in HP-130 and S699. In DNA microarray experiments the two strains were cultivated under standard batch-culture conditions in modified RFB 2244 fermentation medium, and samples were collected at different time points (24-72 h) for RNA extraction. Gene expression data were analyzed to identify transcripts modulated during the growth curve. Amycolatopsis mediterranei S699 (WT) and the improved rifamycin B producer (HP-130) were profiled at 5 time points. For each time point two biological replicates are provided.

Dataset Information

Proteomic analyses reveal metabolic responses to 3-hydroxypropionic acid synthesized internally in Synechocystis sp. PCC 6803

Publications

Proteomic and metabolomic analyses reveal metabolic responses to 3-hydroxypropionic acid synthesized internally in cyanobacterium <i>Synechocystis</i> sp. PCC 6803.

Similar Datasets

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