Identification of transporter proteins for PQQ-secretion pathways by transcriptomics and proteomics analysis in Gluconobacter oxydans WSH-003

Wan, Hui; Xia, Yu; Li, Jianghua; Kang, Zhen; Zhou, Jingwen

doi:10.1007/s11705-016-1580-4

Cited by 21 publications

(14 citation statements)

References 47 publications

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“…First, we reacted PqqB with L-DOPA (lacking the Cys moiety), observing a UV−vis absorbance feature similar to the OH-Cys-DOPA quinone product (Figure 3e); LC-HRMS analyses showed a product mass consistent with OH-DOPA quinone (addition of 14 Da (Figure S8)). Next, tandem MS (MS/MS) analyses were performed on the product of the PqqB reaction with Cys-DOPA under an 16 O 2 atmosphere. Collision-induced dissociation (CID) of the hydroxylated Cys-DOPA (m/z = 331.0596, [M + H] + ) produces a major fragment ion at m/z = 313.05 ([M + H] + ) ascribed to a neutral loss of a molecule of water (−18 Da) (Figure 4b).…”

Section: ■ Results and Discussionmentioning

confidence: 99%

“…Without a conclusive demonstration of robust PqqB activity, proposals of function remain speculative. 8,14,16 Herein, we present a combination of metal and ligand binding studies, X-ray crystallography, and detailed activity assays to show that PqqB is a previously uncharacterized hydroxylase stepwise insertions of oxygen into the tyrosine ring of a PqqAderived cross-linked diamino acid substrate, along with a down-pathway reaction also detected herein, permit a close-tocomplete description of this elusive biosynthetic pathway.…”

Section: ■ Introductionmentioning

confidence: 85%

“…PqqB has been reported to hydrolyze amide and thioester bonds of the substrates of MBL and glyoxalase, respectively; however, the turnover rates for these processes are extremely low and suggest residual ancestral activity. Without a conclusive demonstration of robust PqqB activity, proposals of function remain speculative. ,, Herein, we present a combination of metal and ligand binding studies, X-ray crystallography, and detailed activity assays to show that PqqB is a previously uncharacterized hydroxylase. The aggregate data indicate that PqqB is capable of generating a hydroxyquinone intermediate that possesses the final quinone moiety of the mature PQQ cofactor.…”

Section: Introductionmentioning

confidence: 99%

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Discovery of Hydroxylase Activity for PqqB Provides a Missing Link in the Pyrroloquinoline Quinone Biosynthetic Pathway

et al. 2019

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Understanding the biosynthesis of cofactors is fundamental to the life sciences, yet to date a few important pathways remain unresolved. One example is the redox cofactor pyrroloquinoline quinone (PQQ) which is critical for C1 metabolism in many microorganisms, a disproportionate number of which are opportunistic human pathogens. While the initial and final steps of PQQ biosynthesis, involving PqqD/E and PqqC, have been elucidated, the precise nature and order of the remaining transformations in the pathway are unknown. Here we show evidence that the remaining essential biosynthetic enzyme PqqB is an iron-dependent hydroxylase catalyzing oxygen insertion reactions that are proposed to produce the quinone moiety of the mature PQQ cofactor. The demonstrated reactions of PqqB are unprecedented within the metallo β-lactamase protein family and expand the catalytic repertoire of non-heme iron hydroxylases. These new findings also generate a nearly complete description of the PQQ biosynthetic pathway.

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Section: ■ Results and Discussionmentioning

confidence: 99%

Section: ■ Introductionmentioning

confidence: 85%

Section: Introductionmentioning

confidence: 99%

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Discovery of Hydroxylase Activity for PqqB Provides a Missing Link in the Pyrroloquinoline Quinone Biosynthetic Pathway

et al. 2019

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“…Here we would like to mention six Chinese groups from which you could get a common sense of the main research group on Synbio in China. They are Prof. Yang's group of Shanghai Institutes for Biological Sciences [20], Prof. Zhou's group of Jiangnan University [21], Prof. Xiao's group of Tianjin University [22], Prof. Su's group of Beijing University of Chemical Technology [23], Prof. Dai's group of Tsinghua University before 2018 (now of Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences [24]) and Prof. Yu's group of Zhejiang University [25].…”

Section: Bio-and Medical Are the New Trendsmentioning

confidence: 99%

Steps of fronts in chemical engineering: An overview of the publications of FCSE

Zhu

Huang

Wang

2018

Front. Chem. Sci. Eng.

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“…Liao et al (2015) identified a strong promoter, P r2 , from the RNA-Seq data of Bacillus amyloliquefaciens and verified it by measuring betagalactosidase activity. Several studies about the transcriptome analysis of G. oxydans has been reported to reveal the secretion pathways of PQQ (Wan et al, 2017) and the response to osmotic and oxidative stress of 2-keto-L-gulonic acid (2-KLG) (Fang et al, 2020). Kranz et al (2018) provided deep insights into the transcriptional landscapes of G. oxydans including promoters and other regulatory elements.…”

Section: Introductionmentioning

confidence: 99%

Identification of Gradient Promoters of Gluconobacter oxydans and Their Applications in the Biosynthesis of 2-Keto-L-Gulonic Acid

Chen

Liu

et al. 2021

Front. Bioeng. Biotechnol.

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The acetic acid bacterium Gluconobacter oxydans is known for its unique incomplete oxidation and therefore widely applied in the industrial production of many compounds, e.g., 2-keto-L-gulonic acid (2-KLG), the direct precursor of vitamin C. However, few molecular tools are available for metabolically engineering G. oxydans, which greatly limit the strain development. Promoters are one of vital components to control and regulate gene expression at the transcriptional level for boosting production. In this study, the low activity of SDH was found to hamper the high yield of 2-KLG, and enhancing the expression of SDH was achieved by screening the suitable promoters based on RNA sequencing data. We obtained 97 promoters from G. oxydans’s genome, including two strong shuttle promoters and six strongest promoters. Among these promoters, P3022 and P0943 revealed strong activities in both Escherichia coli and G. oxydans, and the activity of the strongest promoter (P2703) was about threefold that of the other reported strong promoters of G. oxydans. These promoters were used to overexpress SDH in G. oxydans WSH-003. The titer of 2-KLG reached 3.7 g/L when SDH was under the control of strong promoters P2057 and P2703. This study obtained a series of gradient promoters, including two strong shuttle promoters, and expanded the toolbox of available promoters for the application in metabolic engineering of G. oxydans for high-value products.

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Identification of transporter proteins for PQQ-secretion pathways by transcriptomics and proteomics analysis in Gluconobacter oxydans WSH-003

Cited by 21 publications

References 47 publications

Discovery of Hydroxylase Activity for PqqB Provides a Missing Link in the Pyrroloquinoline Quinone Biosynthetic Pathway

Discovery of Hydroxylase Activity for PqqB Provides a Missing Link in the Pyrroloquinoline Quinone Biosynthetic Pathway

Steps of fronts in chemical engineering: An overview of the publications of FCSE

Identification of Gradient Promoters of Gluconobacter oxydans and Their Applications in the Biosynthesis of 2-Keto-L-Gulonic Acid

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