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( Hae-gwang Hwang ),( Dae-hwan Kim ),( Jeongmin Lee ),( Youngwon Mo ),( Se-hoon Lee ),( Yongjin Lee ),( Jae Wook Hyeon ),( Sol Moe Lee ),( Yong-pil Cheon ),( Eun-kyoung Choi ),( Su Yeon Kim ),( Yeong 한국미생물생명공학회(구 한국산업미생물학회) 2018 Journal of microbiology and biotechnology Vol.28 No.10
Recombinant (rec) prion protein (PrP) is an extremely useful resource for studying protein misfolding and subsequent protein aggregation events. Here, we report mass production of high-purity rec-polypeptide encoding the C-terminal globular domain of PrP; (90-230) for human and (89-231) for murine PrP. These proteins were expressed as His-tagged fusion proteins in E. coli cultured by a high cell-density aerobic fermentation method. RecPrPs recovered from inclusion bodies were slowly refolded under reducing conditions. Purification was performed by a sequence of metal-affinity, cation-exchange, and reverse-phase chromatography. The current procedure yielded several dozens of milligrams of recPrP per liter with >95% purity. The purified recPrPs predominantly adopted an α-helix-rich conformation and were functionally sufficient as substrates to measure the seeding activity of human and animal prions. Establishment of a procedure for high-level production of highpurity recPrP supports the advancement of in vitro investigations of PrP including diagnosis for prion diseases.
Recombinant Glargine Insulin Production Process Using Escherichia coli
( Hae-gwang Hwang ),( Kwang-jin Kim ),( Se-hoon Lee ),( Chang-kyu Kim ),( Cheol-ki Min ),( Jung-mi Yun ),( Su Ui Lee ),( Young-jin Son ) 한국미생물 · 생명공학회 2016 Journal of microbiology and biotechnology Vol.26 No.10
Glargine insulin is a long-acting insulin analog that helps blood glucose maintenance in patients with diabetes. We constructed the pPT-GI vector to express prepeptide glargine insulin when transformed into Escherichia coli JM109. The transformed E. coli cells were cultured by fed-batch fermentation. The final dry cell mass was 18 g/l. The prepeptide glargine insulin was 38.52% of the total protein. It was expressed as an inclusion body and then refolded to recover the biological activity. To convert the prepeptide into glargine insulin, citraconylation and trypsin cleavage were performed. Using citraconylation, the yield of enzymatic conversion for glargine insulin increased by 3.2-fold compared with that without citraconylation. After the enzyme reaction, active glargine insulin was purified by two types of chromatography (ion-exchange chromatography and reverse-phase chromatography). We obtained recombinant human glargine insulin at 98.11% purity and verified that it is equal to the standard of human glargine insulin, based on High-performance liquid chromatography analysis and Matrixassisted laser desorption/ionization Time-of-Flight Mass Spectrometry. We thus established a production process for high-purity recombinant human glargine insulin and a method to block Arg (B31)-insulin formation. This established process for recombinant human glargine insulin may be a model process for the production of other human insulin analogs.
A New Rose Cultivar, "Fine Pink" with Pink Color Flower and Resistance to the Powdery Mildew
Gwang Yeon Gi,Kyung Cheuol Cho,Taek Sang Na,Ya Song Lee,Myung Su Cho,Mi Hyang Jang,In Taek Hwang,Joung Keun Kim,Hae Ryoung Shin,Tae Ho Han 한국육종학회 2007 한국육종학회지 Vol.39 No.4
A new pale pink colored standard rose (Rosa hybrida Hort.) cultivar, 'Fine Pink' was developed at the Jeollanamdo Agricultural Research and Extention Services (JARES). Seedling and line selection were conducted in 2003 and its characteristics were examined three times from 2004 to 2006. 'Fine Pink' was obtained by crosse between 'Nobless' and 'Yvessaaintlaurent'. It shows a red group color (RHS No. 56C), very weak fragrance, and standard flower with good harmony between ray floret and flower center. It has a characteristic of straight stem. The average yield during 2004-2006 was 113 stems/m2. It has 10 days vase life and resistance to powdery mildew. This cultivar was registered as a commercial cultivar in 2007. It is suitable for greenhouse in all regions in Korea and will be released to farmers in 2009.