Neuregulin 4 alleviates hepatic steatosis via activating AMPK/mTOR-mediated autophagy in aged mice fed a high fat diet.

Neuregulin 4 (Nrg4) is a brown fat-enriched endocrine factor that exerts beneficial metabolic effects on insulin resistance and hepatic steatosis. Autophagy is a mechanism that is essential for preventing hepatic steatosis. The aim of this study was to explore whether Nrg4 ameliorates hepatic steatosis by inducing autophagy. Aged C57BL/6 mice were maintained on a high fat diet with or without Nrg4 intervention for 3 months. Lipid accumulation in the liver was investigated. Autophagy related protein levels along with related signaling pathways that regulate autophagy were evaluated. In addition, the effects of Nrg4 on autophagy were also determined in cultured L-02 cells. Nrg4 decreased high-fat induced intrahepatic lipid content both in vivo and in vitro. Autophagy level in the liver also decreased in obese mice and Nrg4 intervention reactivated autophagy. Further, Nrg4 intervention was found to have activated autophagy via the adenosine monophosphate-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) signaling pathway. Moreover, when the AMPK/mTOR pathway was suppressed or autophagy was inhibited, the beneficial effects of Nrg4 intervention on hepatic steatosis were diminished. These results indicated that Nrg4 intervention attenuated hepatic steatosis by promoting autophagy in the liver of aged obese mice. Additionally, Nrg4 induced autophagy via the AMPK/mTOR signaling pathway.

Desired soy sauce characteristics and autolysis of Aspergillus oryzae induced by low temperature conditions during initial moromi fermentation.

This is the first report on the effect of low temperature stress applied during initial moromi fermentation on the quality and taste of soy sauce. Koji was prepared to yield initial moromi under three comparative fermentation conditions over 9 days: (1) 4 °C and 0% brine (i.e., water) (LTSF); (2) 4 °C and 16% w/w brine (LTSH); (3) 25 °C and 16% w/w brine (the control, CRTH). Greater extent of autolysis in samples was found under low temperature stress conditions (i.e., at 4 °C, a temperature much lower than the normal temperature range like 25 °C for natural microbial growth and performance). Compared to CRTH, LTSF had a two-fold increase of glutaminase activity in dregs and 65.17% increase in supernatant, and after 60 days of moromi fermentation, a 5.73% and 3.47% increase, respectively, in the contents of glutamic acid (Glu) and aspartic acid (Asp). LTSF had the highest total free amino acid content due to both the low temperature stress and absence of salt. The intensity ranking of umaminess and kokumi sensation (LTSF > LTSH > CRTH) revealed by sensory analysis followed the changing trends of their umami and sweet amino acid contents with a trend reversal in the bitter amino acid content. Low temperature (4 °C) without brine for initial moromi fermentation seemed beneficial, leading to a soy sauce product with desired taste and amino acid contents.

Modification of soy protein isolate by glutaminase for nanocomplexation with curcumin.

Soy protein isolate (SPI) was first treated with glutaminase to yield modified SPI (E-SPI) before its complexation with curcumin. Comparisons were made between SPI and E-SPI concerning their characteristics and effectiveness in complexing with curcumin, along with changes in physicochemical properties, 2,2-diphenyl-1-picryhydrazyl (DPPH)-scavenging capacity and storage stability of curcumin upon complexation. The action of glutaminase did not markedly change SPI structure, but modified some secondary structures, causing greater protein unfolding with more hydrophobic clusters and amino acids exposed. Glutaminase treatment increased DPPH-scavenging capacity, foaming capacity and stability of SPI. The protein concentration played a role in the changes induced by glutaminase treatment and complexation with curcumin, and 1% protein seemed beneficial (highest DPPH scavenging activity; highest loading amount 10.07%; 98% and >90% curcumin retained in 1%-E-SPI at 4 °C for 2 h and 12 h, respectively). Therefore, glutaminase treatment of SPI before complexation with curcumin appeared to be appropriate.

Synthesis and Sensory Characteristics of Kokumi γ-[Glu]n-Phe in the Presence of Glutamine and Phenylalanine: Glutaminase from Bacillus amyloliquefaciens or Aspergillus oryzae as the Catalyst.

The transpeptidase activity of glutaminase from Bacillus amyloliquefaciens (GBA) and Aspergillus oryzae (GAO) to yield γ-[Glu]n-Phe peptides were verified for the first time. In the presence of Gln and Phe, γ-Glu-Phe and γ-Glu-γ-Glu-Phe were synthesized by GAO, and γ-Glu-Phe, γ-Glu-γ-Glu-Phe, γ-Glu-γ-Glu-γ-Glu-Phe, γ-Glu-γ-Glu-γ-Glu-γ-Glu-Phe, and γ-Glu-γ-Glu-γ-Glu-γ-Glu-γ-Glu-Phe were synthesized by GBA. The Km values for the transpeptidation catalyzed by GBA and GAO were 47.88 and 153.92 mM (Phe as the acceptor), 84.89 and 236.47 mM (γ-Glu-Phe as the acceptor), indicating that GBA had a greater affinity than GAO for Phe and γ-Glu-Phe in the transpeptidation reaction. The Km values for the transpeptidation catalyzed by GBA against acceptors, Phe and γ-[Glu](1≤n<5)-Phe (47.88-206.47 mM), increased with an elevated number of γ-glutamyl residue within the acceptor. The optimal conditions for γ-[Glu]n-Phe synthesis were pH 10 and 37 °C for 3 h, 300 mM Gln, 100 mM Phe, 0.05 U/mL GBA. All the γ-[Glu](1≤n≤5)-Phe exhibited astringency in water and imparted a kokumi taste to commercial soy sauce and model chicken broth. The astringent threshold values (2.5-3.92 mM) were approximately 3-fold of the kokumi threshold concentrations (0.78-1.53 mM). γ-[Glu]n-Phe or the post-enzymatic reaction mixture enhanced the umami intensity of commercial soy sauce and model chicken broth.