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Expression, Purification, and Bioactivity of (GLP-1A2G)2-HSA Analogs in Pichia pastoris GS115
Wenfang Dou,Junshang Feng,Xiaomei Zhang,Hongyu Xu,Jinsong Shi,Zhenghong Xu 한국생물공학회 2013 Biotechnology and Bioprocess Engineering Vol.18 No.6
We developed (GLP-1A2G)2-HSA (GGH) analogsthat are resistant to degradation and also show high serumglucose-reducing activity in vivo. Five analogs with Nterminalextensions were designed based on the proteinGGH. Next, we constructed recombinant plasmids capableof expressing the five analogs in methylotrophic yeastPichia pastoris GS115. Expression reached 150 mg/L in asmall-scale incubation. Fusion proteins were successfullypurified from the supernatant using ultrafiltration concentration,affinity absorption chromatography, hydrophobicchromatography, ion exchange chromatography and gelfiltration. A single band was observed on SDS-PAGE andthe purity was 97%. Activity test results suggested thatboth A-GGH and G-GGH showed better activity in vitroand that their cAMP levels were significantly increased by10-fold compared to GGH without N-terminal extension. Additionally, A-GGH efficiently enhanced the glucoseloweringeffect, which was maintained after the administrationfor 24 h. A-GGH is a potential drug for treating type 2diabetes.
Ning Dong,Jianxing Zhou,Ruiting Tong,Quanwei Cui,Yiyi Kong,Junshang Feng,Shichen Xiao 대한기계학회 2023 JOURNAL OF MECHANICAL SCIENCE AND TECHNOLOGY Vol.37 No.12
Wear of tooth profiles and thermal deformation caused by temperature changes are difficult to avoid in gear systems and have important effects on the performance, even to failure. In this paper, a dynamic wear prediction model with coupled thermal deformation and wear is proposed to study the interaction between wear and dynamics of planetary gear system under temperature field, accounting for the effect of temperature on material properties. The system's performance under different wear cycles and temperatures is analyzed. The results show that the difference in the structure makes the curve shape of the thermal deformation error for the external and internal gear pairs completely different, which in turn affects the dynamic response. The distribution and the degree of fluctuation of tooth profile wear depth change as the mesh cycle increases. The early wear is beneficial to the system's uniform load performance and vibration, and the performance decreases as the wear increases, but the external and internal gear pairs are affected by wear to a different extent. The thermal deformation and wear errors have competing or facilitating mechanisms depending on the temperature difference and the degree of wear.