FOOD CONTROL
·
2023
·
IF 6
Shiqi Hu,Xinglian Xu,Wangang Zhang,Chunbao Li,Guanghong Zhou
The activity of cathepsin B in Jinhua ham was affected by various processing parameters, which were investigated and quantified using a multi-layer artificial neural network (ANN). The key factors that influence ham quality are processing temperature, pH, and sodium chloride content. The study discussed the univariate analysis and multivariate interactions between two or three factors. The established ANN model was used for further investigation, and the actual enzyme activity of cathepsin B was characterized. The results showed that high temperature (greater than 60 °C) and an alkaline environment or the addition of Na + cation significantly reduced the enzyme activity ( p < 0.05). The optimal conditions for protease activity were found to be at 40 °C and pH 6.0. The enzyme was found to be unstable at a temperature of 20 °C or pH 6.5–7.0 in the presence of NaCl, indicating significant interactions between temperature, pH value and NaCl content ( p < 0.01). Furthermore, the ANN model showed good match between true and predicted values. The correlation coefficient was 0.98468, and the model provided supplementary information about the effect of NaCl on cathepsin B activity within 0.4M–0.6M. The findings could be useful in Jinhua ham production and the study of the structure-activity relationship of cathepsin B.
研究领域:
食品科学
·
酶学
·
生物化学
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产品关联
DOI
高影响因子
NEUROTOXICOLOGY
·
2026
·
IF 3.8
Yongke Yuan,Mengchen Liu,Mengdi Zhai,Meijia Wu,Wenhui Zhang,Yongjuan Xin,Yue Ba,Yu Zhang,Lin Han,Lihua Gao,Hui Huang
Environmental lead (Pb) exposure and high-fat diet (HFD) intake are known to trigger massive neuronal apoptosis that contributes to neurologic dysfunction. Previous studies mainly focused on isolated mitochondrial dysfunction, autophagy-lysosomal pathway damage, and neuronal apoptosis from Pb or HFD. Nevertheless, the precise role of the lysosomal and proteasomal clearance processes in promoting neuronal cell death induced by Pb and HFD remains unclear. In this study, male Sprague-Dawley (SD) rats were employed to elucidate the mechanisms underlying Pb-induced neurotoxic mitochondrial apoptosis and to investigate the effects of co-exposure to HFD. In vitro , PC12 cells were treated with Pb and palmitic acid (PA) to mimic the in vivo conditions. Specifically, such exposures prompted the translocation of cathepsin B (CTSB) from lysosomes to the cytosol and downregulation of lysosomal-associated membrane protein 1 (LAMP1), ATPase H⁺ Transporting V1 Subunit A (ATP6V1A), and Bcl-2 within rat cortex. In contrast, the levels of pro-apoptotic factors including BID, Bax, mitochondrial cytochrome C (Cyt C), and caspase 3 were significantly elevated. We also observed a significant decrease in the fluorescence intensity of lysosomes while lysosomal pH was ascertained to have ascended appreciably. Concurrently, CTSB inhibition by CA-074 me prevented neural apoptosis processing triggered by Pb and/or PA via mitigating lysosomal damage and mitochondrial Cyt C translocation, indicating that CTSB is a key mediator in the apoptotic process induced by Pb and HFD.
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DOI
PubMed
高影响因子
LWT-FOOD SCIENCE AND TECHNOLOGY
·
2023
·
IF 6
Shiqi Hu,Guanghong Zhou,Xinglian Xu,Wangang Zhang,Chunbao Li
To produce the high quality products and improve the stability of Jinhua hams, the activities of cathepsins and their correlation with processing parameters were explored. The effect of heat treatment on the structure of cathepsin B, together with potential of hydrogen and NaCl concentration, were investigated. At 45 °C, the activity of cathepsin B was maximum and it decreased rapidly at high temperature. The decrease on sulfhydryl group content indicated that the enzyme activity varied due to the breaking of disulfide bond in molecule. Infrared spectroscopy indicated that the secondary structure of cathepsin B had altered. Meanwhile, the root mean squared deviation and the root mean square fluctuation curves reflected that the cathepsin B vitality was in a decline under high temperature. In addition, in the presence of 0.6 mol ⋅ L −1 NaCl, lots of conformational changes were evidenced. Under acidic conditions (pH = 6), there were more formed hydrogen bonds, and the hydrogen bonding interactions became stronger. Simulation results contributed to providing an important theoretical basis for understanding the roles of processing parameters on the quality control of Jinhua ham.
研究领域:
食品科学
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生物化学
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产品关联
DOI
高影响因子
MICROCHIMICA ACTA
·
2023
·Vol. 190
·期 5
·1-9
·
IF 5.7
Li Xinxin,Zhang Meiling,Zhang Hua,Wang Zhenxin,Zhang Huimao
A simple fluorescence resonance energy transfer (FRET) sensing platform (termed as USP), comprised of upconversion nanoparticles (UCNPs) as the energy donor and Cy5 as the energy acceptor, has been synthesized for cathepsin B (CTSB) activity detection in vitro and in vivo. When Cy5-modified peptide substrate (peptide-Cy5) of CTSB is covalently linked on the surface of UCNPs, the FRET between the UCNPs (excitation: 980 nm; emission: 541 nm/655 nm) and Cy5 (excitation: 645 nm) leads to a reduction in the red upconversion luminescence (UCL) signal intensity of UCNPs. Cy5 can be liberated from UCNPs in the presence of CTSB through the cleavage of peptide-Cy5 by CTSB, leading to the recovery of the red UCL signal of UCNPs. Because the green UCL signal of UCNPs remains constant during the CTSB digestion, it can be considered as an internal reference. The findings demonstrate the ability of USP to detect CTSB with the linear detection ranges of 1 to 100 ng·mL −1 in buffer and 2 × 10 3 to 1 × 10 5 cells in 0.2 mL cell lysates. The limits of detection (LODs) are 0.30 ng·mL −1 in buffer and 887 cells in 0.2 mL of cell lysates (S/N = 3). The viability of USP to detect CTSB activity in tumor-bearing mice is has further been investigated using in vivo fluorescent imaging. Graphical abstract
研究领域:
生物技术
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生物化学
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分子生物学
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癌症研究
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纳米技术
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）