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

Author:
Jin, Feng-Jie, Han, Pei, Zhuang, Miao, Zhang, Zhi-Min, Jin, Long, Koyama, Yasuji
Source:
Applied microbiology and biotechnology 2018 v.102 no.1 pp. 14-332
ISSN:
0175-7598
Subject:
Aspergillus oryzae, alpha-glucosidase, alpha-mannosidase, beta-galactosidase, carbohydrate metabolism, carbon metabolism, genes, liquids, mammals, manufacturing, microorganisms, phenotype, protein degradation, protein synthesis, proteome, proteomics, reverse transcriptase polymerase chain reaction, transcription factors
Abstract:
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 fermentative manufacturing and commercial enzyme production in A. oryzae because of its effect on growth. 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. A total of 5447 proteins were identified, and 568 were differentially expressed proteins (DEPs). Of the DEPs, 251 proteins were increased by 1.5-fold, and 317 proteins were decreased by 1.5-fold in an sclR-disrupted strain compared to the control. The comparison of the quantitative TMT results revealed that SclR was mainly involved in carbon metabolism, especially carbohydrate metabolism. In addition, an enzyme profile by a semi-quantitative method (API-ZYM) indicated that three enzymes (β-galactosidase, α-glucosidase, and α-mannosidase) were significantly less active in the ∆sclR strain than in the control. Moreover, quantitative RT-PCR showed that the expression of certain genes was changed similarly to their corresponding proteins. These results suggested that a possible function of SclR during growth of A. oryzae is its important involvement in carbohydrate metabolism.