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Purification and properties of a milk-clotting enzyme produced by Bacillus amyloliquefaciens D4
Xiaoling He,Bozhong Gan,Fazheng Ren,Huiyuan Guo,Weibing Zhang,Xi Song 한국화학공학회 2011 Korean Journal of Chemical Engineering Vol.28 No.1
The milk-clotting enzyme from Bacillus amyloliquefaciens D4 was purified to 17.2-fold with 20% recovery by precipitation in ammonium sulfate and ion-exchange chromatography. The molecular mass of the enzyme was 58.2kDa as determined by SDS-PAGE, and it was proved to be a metalloprotease by EDTA inhibition. The enzyme was active in the pH range 5.5-7.0 and was inactivated completely by heating at 55℃ for 20 min. The highest level of enzyme activity was obtained at 65 ℃, pH 5.5, in the presence of 25mM CaCl_2. The milk-clotting activity was inhibited only slightly by Na^+ and K^+ and significantly by Cu^(2+), Zn^(2+) and Sn^(2+). The Km value of this enzyme was 0.471 mg/mL. The high level of milk-clotting activity coupled with a low level of thermal stability suggested that the milk-clotting enzyme from B. amyloliquefaciens D4 should be considered as a potential substitute for calf rennet.
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Xiaoling Wang,Hai Lu,Tiehong Song,Ke Zhao 한국화학공학회 2019 Korean Journal of Chemical Engineering Vol.36 No.3
The experimental results and material balance analysis in this paper revealed the regularity of poly-hydroxy alkanoates (PHA) and total phosphorus (TP) metabolism in a continuous-flow single-sludge wastewater treatment system under different main anoxic section oxidation-reduction potential (ORPan) conditions. We also evaluated the effectiveness of the operation control parameters of ORPan as the continuous-flow single-sludge sewage treatment system from the aspect of the reaction mechanism. Using a programmable logic controller (PLC) automatic control system to take the circulating flow in nitrification as the controlled variable based on the feedback control structure, an experimental study was carried out under the condition of ORPan setting value of 143mV, 123mV, 105mV, 95mV, 72 mV and 57mV, respectively, with other operational design parameters remaining unchanged. Influent water quality of chemical oxygen demand/total nitrogen (COD/TN) was 5.0±0.6. The results showed that when ORPan was set at 95mV, the maximum values of PHA synthesis and storage rate, PHA degradation rate, phosphorus release rate and phosphorus absorption rate in anaerobic and pre-anoxic segments were 82.34, 7.90, 47.31, 14.27, 1.50 and 8.52mg/ (L·h), respectively. According to the metabolic mechanism of PHA and TP, ORPan was further proved to be the operation control parameter of the continuous-flow single-sludge sewage treatment system, and when the COD/TN value was 5.0±0.6, the optimal setting value was 95mV.
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Lai Yuting,Hu Li,Yang Lu,Hu Xianting,Song Xiaole,Yang Jingyi,Li Hongbin,Chen Kun,Li Huabin,Wang Dehui 대한천식알레르기학회 2021 Allergy, Asthma & Immunology Research Vol.13 No.5
Purpose: Serum/glucocorticoid-regulated kinase 1 (SGK1) has recently emerged as a critical regulator of inflammatory diseases. In this study, we examined SGK1 expression and its possible pathogenic roles in chronic rhinosinusitis (CRS). Methods: Immunohistochemistry, western blotting, Bio-Plex assay, enzyme-linked immunosorbent assays, and quantitative real-time polymerase chain reaction were performed to assess protein and gene expression levels. The mRNA expression levels of SGK1 and interleukin-6 (IL-6) were extracted from a CRS database to perform correlation analysis. Stable cell lines with SGK1 overexpression (16HBE) and knockdown (A549) were constructed to investigate the interaction between SGK1 and IL-6 in vitro. Results: SGK1 exhibited strong cytoplasmic and nuclear staining in the epithelial layers and the lamina propria of nasal polyps (NPs) and in the mucosal tissues of CRS without nasal polyps (CRSsNP). The mRNA and protein expression levels of SGK1 and IL-6 were significantly increased in NPs and CRSsNP tissues, compared to control tissues. SGK1 phosphorylation was significantly greater in NPs than in CRSsNP tissues (P < 0.01). The mRNA levels of SGK1 and IL-6 were significantly correlated (P < 0.001, r = 0.649). Exposure to IL-6 significantly increased SGK1 expression in cultured dispersed NP cells, 16HBE cells, and A549 cells. IL-6 expression was significantly down-regulated in SGK1-overexpressing 16HBE cells (P < 0.01) and significantly up-regulated in SGK1-knockdown A549 cells (P < 0.05). Administration of GSK650394, a SGK1 inhibitor, significantly increased IL-6 self-induced mRNA expression in cultured dispersed NP cells and 16HBE cells. Conclusions: The interaction between SGK1 and IL-6 may play an anti-inflammatory role in IL-6-induced inflammation in the pathogenesis of CRS.
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AUF1 promotes let-7b loading on Argonaute 2
Yoon, Je-Hyun,Jo, Myung Hyun,White, Elizabeth J.F.,De, Supriyo,Hafner, Markus,Zucconi, Beth E.,Abdelmohsen, Kotb,Martindale, Jennifer L.,Yang, Xiaoling,Wood III, William H.,Shin, Yu Mi,Song, Ji-Joon,T Cold Spring Harbor Laboratory Press 2015 Genes & development Vol.29 No.15
<P>Yoon et al. discovered that RBP AU-rich-binding factor 1 (AUF1) promotes let-7b loading onto Argonaute 2 (AGO2), the catalytic component of the RNA-induced silencing complex (RISC). In turn, AGO2–let-7 triggered target mRNA decay.</P><P>Eukaryotic gene expression is tightly regulated post-transcriptionally by RNA-binding proteins (RBPs) and microRNAs. The RBP AU-rich-binding factor 1 (AUF1) isoform p37 was found to have high affinity for the microRNA let-7b in vitro (<I>K</I><SUB>d</SUB> = ∼6 nM) in cells. Ribonucleoprotein immunoprecipitation, in vitro association, and single-molecule-binding analyses revealed that AUF1 promoted let-7b loading onto Argonaute 2 (AGO2), the catalytic component of the RNA-induced silencing complex (RISC). In turn, AGO2–let-7 triggered target mRNA decay. Our findings uncover a novel mechanism by which AUF1 binding and transfer of microRNA let-7 to AGO2 facilitates let-7-elicited gene silencing.</P>
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