Identification and functional characterization of glycerol dehydrogenase reveal the role in kojic acid synthesis in Aspe
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(2020) 36:136
ORIGINAL PAPER
Identification and functional characterization of glycerol dehydrogenase reveal the role in kojic acid synthesis in Aspergillus oryzae Junxia Fan1 · Zhe Zhang1 · Chuannan Long1 · Bin He1 · Zhihong Hu1 · Chunmiao Jiang1 · Bin Zeng1 Received: 2 May 2020 / Accepted: 6 August 2020 © Springer Nature B.V. 2020
Abstract Glycerol dehydrogenase has been identified and characterized functionally in many species. However, little is known about glycerol dehydrogenase genes and their functions in Aspergillus oryzae. Here, a total of 45 glycerol dehydrogenase genes in Aspergillus oryzae were identified and renamed from AoGld1 to AoGld45 according to their chromosome distribution. They were classified into three groups based on phylogenetic analysis. Synteny analysis revealed that thirteen AoGld genes are conserved among Aspergillus species. Promoter analysis displayed that AoGld3 and AoGld13 harbored multiple binding elements of GATA-type transcription factors and zinc-finger protein msnA that were involved in nitrogen and kojic acid metabolism, respectively. Moreover, the AoGld3 deletion strain Δgld3 was generated by the CRISPR/Cas9 system, which had no visible growth defects compared with the control wild-type strain under the control and osmotic stress treatments. However, disruption of AoGld3 led to the inhibition of kojic acid production, and the expression of kojA, kojR was downregulated in the Δgld3 strain. Furthermore, when kojA or kojR was overexpressed in the Δgld3 strain, the yield of kojic acid was restored, suggesting that AoGld3 is involved in kojic acid production through affecting the expression of kojR and kojA. Taken together, these findings provide new insights into our understanding of glycerol dehydrogenase and establish foundation for further study of their roles in Aspergillus oryzae. Keywords Kojic acid · Glycerol dehydrogenase · Aspergillus oryzae · Secondary metabolism
Introduction Glycerol is one of the most predominant cryoprotectants, which is used by microorganisms to guard against various stresses, such as osmotic stress (Redkar et al. 1995; Robinson 2001; Williams et al. 2017). The synthesis of glycerol has been well described in the yeast Saccharomyces Electronic supplementary material The online version of this article (https://doi.org/10.1007/s11274-020-02912-4) contains supplementary material, which is available to authorized users. * Zhe Zhang [email protected] * Bin Zeng [email protected] 1
Jiangxi Key Laboratory of Bioprocess Engineering and Co‑Innovation Center for In‑Vitro Diagnostic Reagents and Devices of Jiangxi Province, College of Life Sciences, Jiangxi Science and Technology Normal University, Nanchang 330013, China
cerevisiae, containing two pathways, anabolic glycerol3-phosphate (G3P) pathway and anabolic dihydroxyacetone (DHA) pathway (Nevoigt and Stahl 1997; Norbeck and Blomberg 1997). The anabolic G3P pathway starts with the reduction of dihydroxyacetone phosphate (DHAP) to G3P by a NAD+-dependent glycerol-3-phosphate dehydroge
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