A metabolic regulation method was proposed to increase the microalgal intracellular nicotinamide adenine dinucleotide phosphate (NADPH) by lowering NADPH consumption in the Calvin cycle or accelerating NADPH regeneration in photosynthesis, thereby to promote the asymmetric reduction of prochiral ketone to chiral alcohol catalyzed by microalgae cell. The NADPH consumption was efficiently decreased by inhibiting glyceraldehyde 3-phosphate dehydrogenase with inhibitors, such as iodoacetic acid and Angeli's Salt, which increased the microalgal intracellular NADPH by 80% and 96%, respectively. Correspondingly, the yields of the asymmetric reduction reaction were enhanced by 12.9% and 63.3%, respectively. On the other hand, NADPH regeneration was accelerated by promoting electron transfer and ferredoxin-NADP+ reductase activity with addition of sodium cholate, sodium acetate and sodium thiosulfate; the intracellular NADPH was increased 73.1%, 60.2% and 70.4%, respectively. The yields of the asymmetric reduction reaction were correspondingly enhanced by 118.4%, 36.7% and 26.2%, respectively. This provides a practical technique to promote microalgae intracellular NADPH amount to enhance asymmetric reduction reaction by photo-biocatalytic processes. © 2016 Curtin University of Technology and John Wiley & Sons, Ltd.

Promotion of the microalgal photo-biocatalytic asymmetric reduction of prochiral ketone by NADPH metabolic regulation: Promotion of Biocatalysis by NADPH Metabolic Regulation

Thyroid hormone (TH) is essential for Paralichthys olivaceus metamorphosis. Exogenous TH treatment induces premature metamorphosis in P. olivaceus larvae and a series of studies have been conducted to identify thyroid hormone-regulated functional genes and microRNAs involved in the metamorphosis of P. olivaceus; however, the proteins involved in this process remain to be fully clarified. In this study, the differential proteomic responses of P. olivaceus larvae to exogenous TH treatment were examined using tandem mass tags (TMT) for quantitation labeling followed by liquid chromatography tandem mass spectrometry (LC-MS/MS). The expression levels of 629 cellular proteins were identified to be significantly affected by TH treatment. The reliability of our TMT-labeled LC-MS/MS analysis was verified by examining the mRNA and protein levels of four selected proteins using quantitative real-time reverse-transcription PCR and western blot analyses. The possible biological significance of these proteins was further investigated by Gene Ontology (GO) enrichment, Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment, and protein-protein interaction analyses. Notably, we identified and described five groups of proteins involved in different important life events that were significantly regulated by exogenous TH treatment. Our study provides an improved understanding of the molecular mechanisms by which TH regulates the metamorphosis of P. olivaceus.

Proteomic variation in metamorphosing Paralichthys olivaceus induced by exogenous thyroid hormone

Glyceraldehyde-3-phosphate dehydrogenase(GAPDH) is a kind of protein constitutively expressed in cells,mainly involved in cellular glycolytic pathway.But in recent years,more and more studies have showed that GAPDH is a multi-functional protein,which not only functions as a glycolytic enzyme,but also gets involved in many subcellular activities,such as catalyzing microtubule polymerization,playing a role in protein phosphorylation modification,participating in membrane fusion and membrane transfer,promoting cell apoptosis,regulating protein expression,being involved in DNA damage repair,acting as transferrin receptor,and so on.This review mainly discusses the structural characteristics of GAPDH and the research progress on its functions.

Xiaohong Huang

Papers

Progress on Research of Functions of Glyceraldehyde-3-phosphate Dehydrogenase