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Immobilization of As(V) in Rhizopus oryzae Investigated by Batch and XAFS Techniques.

ABSTRACT: Arsenic (As) contamination in aqueous solutions has become an increasing public concern due to the immense harm to human health. Herein, bioaccumulation of arsenate (As(V)) by Rhizopus oryzae in aqueous systems was investigated under different environmental conditions, such as different pH's, ionic strengths, mycelia dosages, mycelia growths, and temperatures. The results showed that As(V) could be bioaccumulated efficiently by R. oryzae, and the maximum bioaccumulation capacity of As(V) in R. oryzae was 52.4 mg/g at T = 299 K, which was much higher than that for other biomaterials under similar conditions. R. oryzae generated a higher content of thiol compounds under As(V) stress to immobilize As(V) from aqueous solutions. X-ray absorption near-edge spectroscopy analysis indicated that As(V) was partly reduced to As(III) with increasing contact time, which increased As(V) bioaccumulation in mycelia. In addition, extended X-ray absorption fine structure analysis showed that the As-S complex played an important role in As(V) immobilization by mycelia. This study provided an in-depth investigation of intracellular As speciation and coordination in R. oryzae on the molecular scale, which was crucial to understand the interaction mechanisms of As(V) with fungi during environmental cleanup.

SUBMITTER: Song W

PROVIDER: S-EPMC6044679 | biostudies-literature | 2016 Nov

REPOSITORIES: biostudies-literature

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Immobilization of As(V) in Rhizopus oryzae Investigated by Batch and XAFS Techniques.

Song Wencheng W Wang Xiangxue X Wen Tao T Yu Shujun S Zou Yidong Y Sun Yubing Y Hayat Tasawar T Wang Xiangke X

ACS omega 20161111 5

Arsenic (As) contamination in aqueous solutions has become an increasing public concern due to the immense harm to human health. Herein, bioaccumulation of arsenate (As(V)) by Rhizopus oryzae in aqueous systems was investigated under different environmental conditions, such as different pH's, ionic strengths, mycelia dosages, mycelia growths, and temperatures. The results showed that As(V) could be bioaccumulated efficiently by R. oryzae, and the maximum bioaccumulation capacity of ...[more]

PMID: 30023494

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Project description:BACKGROUND: The analysis of transcriptional levels of the genes involved in protein synthesis and secretion is a key factor to understand the host organism's responses to recombinant protein production, as well as their interaction with the cultivation conditions. Novel techniques such as the sandwich hybridization allow monitoring quantitatively the dynamic changes of specific RNAs. In this study, the transcriptional levels of some genes related to the unfolded protein response (UPR) and central metabolism of Pichia pastoris were analysed during batch and fed-batch cultivations using an X-33-derived strain expressing a Rhizopus oryzae lipase under control of the formaldehyde dehydrogenase promoter (FLD1), namely the alcohol oxidase gene AOX1, the formaldehyde dehydrogenase FLD1, the protein disulfide isomerase PDI, the KAR2 gene coding for the BiP chaperone, the 26S rRNA and the R. oryzae lipase gene ROL. RESULTS: The transcriptional levels of the selected set of genes were first analysed in P. pastoris cells growing in shake flask cultures containing different carbon and nitrogen sources combinations, glycerol + ammonium, methanol + methylamine and sorbitol + methylamine. The transcriptional levels of the AOX1 and FLD1 genes were coherent with the known regulatory mechanism of C1 substrates in P. pastoris, whereas ROL induction lead to the up-regulation of KAR2 and PDI transcriptional levels, thus suggesting that ROL overexpression triggers the UPR. This was further confirmed in fed-batch cultivations performed at different growth rates. Transcriptional levels of the analysed set of genes were generally higher at higher growth rates. Nevertheless, when ROL was overexpressed in a strain having the UPR constitutively activated, significantly lower relative induction levels of these marker genes were detected. CONCLUSION: The bead-based sandwich hybridization assay has shown its potential as a reliable instrument for quantification of specific mRNA species in P. pastoris cells grown in fed-batch cultures. As a proof-of-principle, the influence of the carbon and nitrogen sources, the specific growth rate, as well as the ROL overexpression on the transcriptional levels of a reduced set of bioprocess-relevant genes has been quantitatively studied, revealing that ROL overexpression and secretion seems to trigger the UPR in P. pastoris, resulting in a physiological bottleneck for the production process.

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Immobilization of As(V) in Rhizopus oryzae Investigated by Batch and XAFS Techniques.

Publications

Immobilization of As(V) in <i>Rhizopus oryzae</i> Investigated by Batch and XAFS Techniques.

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