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( Xiao Lan Liu ),( Xi Qun Zheng ),( Peng Zhi Qian ),( Narasimha Kumar Kopparapu ),( Yong Ping Deng ),( Masanori Nonaka ),( Naoki Harada ) 한국미생물 · 생명공학회 2014 Journal of microbiology and biotechnology Vol.24 No.2
A fibrinolytic enzyme was produced by an edible mushroom of Pleurotus ostreatus using submerged culture fermentation. The enzyme was purified from the culture supernatant by applying a combination of freeze-thaw treatment, ammonium sulfate precipitation, hydrophobic interaction, and gel filtration chromatographies. The enzyme was purified by a 147-fold, with a yield of 7.54%. The molecular masses of the enzyme an determined by gel filtration and SDSPAGE were 13.6 and 18.2 kDa, respectively. The isoelectric point of the enzyme was 8.52. It hydrolyzed fibrinogen by cleaving the α and β chains of fibrinogen followed by the γ chains, and also activated plasminogen into plasmin. The enzyme was optimally active at 45°C and pH 7.4. The enzyme activity was completely inhibited by EDTA, whereas protease inhibitors of TPCK, SBTI, PMSF, aprotinin and pepstatin showed no inhibition on its activity. The partial amino acid sequences of the enzyme as determined by Q-TOF2 were ATFVGCSATR, GGTLIHESSHFTR, and YTTWFGTFVTSR. These sequences showed a high degree of homology with those of metallo-endopeptidases from P. ostreatus and Armillaria mellea. The purified enzyme can also be applied as a natural agent for oral fibrinolytic therapy or prevention of thrombosis.
( Gui Zi Ye ),( Min Jiang ),( Jian Li ),( Ke Quan Chen ),( Yong Lan Xi ),( Shu Wen Liu ),( Ping Wei ),( Ping Kai Ouyang ) 한국미생물 · 생명공학회 2010 Journal of microbiology and biotechnology Vol.20 No.8
Actinobacillus succinogenes, a representative succinic-acid-producing microorganism, is seriously inhibited by ammonium ions, thereby hampering the industrial use of A. succinogenes with ammonium-ion-based materials as the pH controller. Therefore, this study isolated an ammonium-ion-tolerant mutant of A. succinogenes using a continuous-culture technique in which all the environmental factors, besides the stress (ammonium ions), were kept constant. Instead of operating the mutant-generating system as a nutrient-limited chemostat, it was used as a nutrientunlimited system, allowing the cells to be continuously cultured at the maximum specific growth rate. The mutants were isolated on agar plates containing the acid-base indicator bromothymol blue and a high level of ammonium ions that would normally kill the parent strain by 100%. When cultured in anaerobic bottles with an ammonium ion concentration of 354 mmol/l, the mutant YZ0819 produced 40.21 g/l of succinic acid with a yield of 80.4%, whereas the parent strain NJ113 was unable to grow. When using NH4OH to buffer the culture pH in a 3.0 l stirred bioreactor, YZ0819 produced 35.15 g/l of succinic acid with a yield of 70.3%, which was 155% higher than that produced by NJ113. In addition, the morphology of YZ0819 changed in the fermentation broth, as the cells were aggregated from the beginning to the end of the fermentation. Therefore, these results indicate that YZ0819 can efficiently produce succinic acid when using NH4OH as the pH controller, and the formation of aggregates can be useful for transferring the cells from a cultivation medium for various industrial applications.
Shun-Feng Cheng,Ling Li,Bo Li,Jing-Cai Liu,Fang-Nong Lai,Yong Zhao,Xi-Feng Zhang,Wei Shen,Lan Li 대한독성 유전단백체 학회 2018 Molecular & cellular toxicology Vol.14 No.2
Backgrounds: Di-(2-ethylhexyl) phthalate (DEHP), as an endocrine-disrupting chemical (EDC), is widely used in plasticizer and other productions. Ubiquitous human exposure to DEHP has been proposed to be a potential risk to public health. Developmental exposure to DEHP could alter epigenetic programming and result in adult-onset disease. Methods: In this study, we investigated whether DEHP exposure to pregnant mice affected epigenetic changes as a result of increase in breast cancer incidence. Results: Our results showed that the expression of total 143 epigenetics-related genes in mammary gland cells, have no significantly altered after short time and low-dose treated with DEHP from 0.5 days post-coitum (dpc) to 3.5 dpc of pregnant mice. DNA methylation status of some neoplastic development genes, such as EGFr, Esr1, Pgr, Fos and Rassf5 also had no obvious change. Conclusion: These finding showed no impact of DEHP on the expressive patterns of epigenetics-related genes and DNA methylation of breast cancer-related genes in pregnant mouse mammary gland cells.