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Aspergillus oryzae LC-MSMS - Comparative proteomic analysis: SclR is importantly involved in carbohydrate metabolism in Aspergillus oryzae

ABSTRACT: We previously identified the bHLH transcription factor SclR and found that the loss of SclR function led to significant phenotypic changes, such as rapid protein degradation and cell lysis in dextrin-polypeptone-yeast extract liquid medium. Therefore, this study presents a comparative assessment at the proteome level of the intracellular differences between an sclR-disrupted strain and a control strain using isobaric tandem mass tag (TMT) labeling for quantification.

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Aspergillus Oryzae Rib40

SUBMITTER: Feng-Jie Jin

LAB HEAD: Feng-Jie Jin

PROVIDER: PXD007829 | Pride | 2017-11-07

REPOSITORIES: Pride

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Comparative proteomic analysis: SclR is importantly involved in carbohydrate metabolism in Aspergillus oryzae.

Jin Feng-Jie FJ Han Pei P Zhuang Miao M Zhang Zhi-Min ZM Jin Long L Koyama Yasuji Y

Applied microbiology and biotechnology 20171102 1

The helix-loop-helix (HLH) family of transcriptional factors is a key player in a wide range of developmental processes in organisms from mammals to microbes. We previously identified the bHLH transcription factor SclR in Aspergillus oryzae and found that the loss of SclR function led to significant phenotypic changes, such as rapid protein degradation and cell lysis in dextrin-polypeptone-yeast extract liquid medium. The result implied that SclR is potentially important in both traditional ferm ...[more]

PMID: 29098410

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Project description:Fungal endo-M-NM-2-mannanases (M-NM-2-mannanases) are widely used as industrial enzymes; however, no transcriptional regulator of M-NM-2-mannanases has been identified in fungi or other eukaryotic cells to date. To identify a transcriptional regulator of M-NM-2-mannanases in Aspergillus oryzae, a gene-disruptant library of transcriptional regulators was screened for mutants exhibiting reduced M-NM-2-mannanase activity by using konjac glucomannan as the substrate, and ManR, a Zn(II)2Cys6 type DNA binding protein was identified. Moreover, a manR-overexpressing strain showed significantly increased M-NM-2-mannanase activity. DNA microarray analysis of the manR-disruptant strain and the manR-overexpressing strain further indicated that when konjac glucomannan is used as the carbon source, ManR positively regulates the gene expression of not only M-NM-2-mannanase, but also the enzymes involved in the degradation of galactomannans and glucomannans such as M-NM-1-galactosidase, M-NM-2-mannosidase, acetylmannan esterase, and M-NM-2-glucosidase. Therefore, we conclude that ManR is a positive regulator of the M-NM-2-mannan utilization system in A. oryzae. manR disruptant, manR-overexpressing strain and A. oryzae RkuptrP2-1M-bM-^HM-^FAF/P (derivative of A. oryzae RIB40) were cultivated in minimal medium containing 1% konjac glucomannan as the sole carbon source. After 6 h cultivation, total RNAs from the mycelia were extracted, and DNA microarray analysis was carried out. The analysis of manR disruptant was conducted with 4 biological replications, the analysis of manR overexpressing strain was conducted with 3 biological replications.

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