Tavasoli Tina, Arjmand Sareh, Ranaei Siadat Seyed Omid, Shojaosadati Seyed Abbas, Sahebghadam Lotfi Abbas
Biotechnology Group, Department of Chemical Engineering, Tarbiat Modares University, Tehran, Iran.
Protein Research Center, Shahid Beheshti University, G.C., Tehran, Iran.
Iran J Biotechnol. 2017 Dec 29;15(4):224-231. doi: 10.15171/ijb.1808. eCollection 2017.
Human alpha 1-antitrypsin (AAT) is a monomeric glycosylated protein; it is the potent inhibitor of a whole range of serine proteases and protects tissues against their destructive effects. The human plasma-derived AAT, which is currently used to augment the AAT level in patients, is limited due to high cost and source limitation. Recombinant production of AAT can be considered as a potential alternative. This study aims to develop and optimize a new chemically defi ned medium based on an elemental analysis of the yeast for an effi cient culture of the recombinant yeast-producing secretory AAT. An elemental analysis of Carbon (C), Hydrogen (H), Nitrogen (N), Sulfur (S); CHNS in its abbreviated form, and metallic elements was performed to determine the exact molecular constituent of the . The medium components were selected according to the obtained formula; they were optimized by the response surface methodology (RSM). The grown yeast cell was measured at the end of 18 h glycerol batch culture. The amounts of AAT production and elastase inhibitory capacity (EIC) were measured at the end of three days' methanol feeding. The optimized medium compositions consist of glycerol (40 g.L), KHPO (24.78 g.L), NaCl, (0.88 g.L), MgSO .7H O (1.95 g.L), (NH ) SO (22.76 g.L), and trace elements (20 mL.L). The presented quadratic models show that KH PO and (NH4) SO, are the most abundant ones in the biomass and have the greatest effect on the cell growth, EIC, and AAT protein production responses. According to the results of this study, it can be concluded that the characterizing cell composition formula could be considered as an appropriate method to design culture media in order to improve cell growth and productivity. Compared to the common chemically defi ned media, FM22 and BSM, production of AAT protein increased by 1.5 and 1.4 times, respectively, in this new medium.
人α1 -抗胰蛋白酶（AAT）是一种单体糖基化蛋白；它是多种丝氨酸蛋白酶的强效抑制剂，可保护组织免受其破坏作用。目前用于提高患者AAT水平的人血浆来源的AAT，由于成本高和来源有限而受到限制。AAT的重组生产可被视为一种潜在的替代方法。本研究旨在基于酵母的元素分析开发并优化一种新的化学成分确定的培养基，以高效培养分泌AAT的重组酵母。对碳（C）、氢（H）、氮（N）、硫（S）；简称为CHNS，以及金属元素进行元素分析，以确定酵母的确切分子组成。根据所得配方选择培养基成分；通过响应面法（RSM）对其进行优化。在18小时甘油分批培养结束时测量生长的酵母细胞。在三天甲醇补料结束时测量AAT产量和弹性蛋白酶抑制能力（EIC）。优化后的培养基组成包括甘油（40 g.L）、KHPO（24.78 g.L）、NaCl（0.88 g.L）、MgSO·7H₂O（1.95 g.L）、(NH₄)₂SO₄（22.76 g.L）和微量元素（20 mL.L）。所呈现的二次模型表明，KH₂PO₄和(NH₄)₂SO₄在生物量中含量最高，对细胞生长、EIC和AAT蛋白生产响应的影响最大。根据本研究结果，可以得出结论，表征细胞组成配方可被视为设计培养基以提高细胞生长和生产力的合适方法。与常用的化学成分确定的培养基FM22和BSM相比，在这种新培养基中AAT蛋白产量分别提高了1.5倍和1.4倍。
