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( Yuanda Song ),( Emily Dilger ),( Jessica Bell ),( William A Barton ),( Xianjun Fang ) 생화학분자생물학회 (구 한국생화학분자생물학회) 2010 BMB Reports Vol.43 No.8
We utilized a mammalian expression system to purify and characterize autotaxin (ATX)/lysophospholipase D, an enzyme present in the blood responsible for biosynthesis of lysophosphatidic acid. The human ATX cDNA encoding amino acids 29-915 was cloned downstream of a secretion signal of CD5. At the carboxyl terminus was a thrombin cleavage site followed by the constant domain (Fc) of IgG to facilitate protein purification. The ATX-Fc fusion protein was expressed in HEK293 cells and isolated from conditioned medium of a stable clone by affinity chromatography with Protein A sepharose followed by cleavage with thrombin. The untagged ATX protein was further purified to essential homogeneity by gel filtration chromatography with a yield of approximately 5 mg/liter medium. The purified ATX protein was enzymatically active and biologically functional, offering a useful tool for further biological and structural studies of this important enzyme. [BMB reports 2010; 43(8): 541-546]
Hameed Ahsan,Hussain Syed Ammar,Nosheen Shaista,Muhammad Zafarullah,Wu Yang,Ullah Samee,Suleria Hafiz Ansar Rasul,Song Yuanda 한국응용생명화학회 2020 Applied Biological Chemistry (Appl Biol Chem) Vol.63 No.3
This study aimed at increasing the stability of heat-labile and pH-sensitive microbial antioxidants by the microencapsulation. Microbial antioxidants from Mucor circinelloides were microencapsulated. The physico-chemical and powder flowing properties of resulting microcapsules were evaluated. The initial safety studies were evaluated by in vivo acute oral toxicity tests. The bio-accessibility of powders vs. extracts was analyzed in in vitro digestion models with further application of microcapsules to model food system. Physico-chemical properties were significantly different (p < 0.0001) for all microcapsules regardless of their non-substantial variations (p > 0.05) in powder flowing properties. The microencapsulation of extract with 5% whey protein hydrogels (WPHG) + 5% pectin ( TA) showed higher retain-ability of polyphenols accompanying low degradation in gastric and intestinal digestion and with no major toxicity signs. The addition of TA microcapsule did not produce any nutritional, physico-chemical, compositional, and nutritional distinctions in cheese. Microencapsulation proved to be appropriate approach for not only protecting the thermo-labile and pH-sensitive microbial antioxidants but also for enhanced bioavailability, and targeted release of bioactive extracts.