%0 Journal Article
%T Influence of methanol concentration on purification recovery of consensus interferon-a produced by Pichia pastoris<br>甲醇浓度对毕赤酵母表达重组人复合α干扰素分离纯化得率的影响
%A Dan Wu
%A Ju Chu
%A Yonghong Wang
%A Yingping Zhuang
%A Siliang Zhang
%A <br>吴丹
%A 储炬
%A 王永红
%A 庄英萍
%A 张嗣良
%J 生物工程学报
%D 2011
%I 
%X We observed the influence of methanol concentration on purification yield rate of recombinant human consensus interferon (cIFN) produced by Pichia pastoris. Fermentations controlled at 0.75% (W/V) methanol showed a 24% increase in cIFN expression compared to the induced culture at 0.25% methanol. However, a sharp drop of 79% in purification yield rate of cIFN was seen at 0.75% methanol compared to 0.25% methanol. To seek the reason, we analyzed the stability of cIFN by SDS-PAGE and Native-PAGE as well as Western blotting. The electrophoresis results revealed that cIFN formed a lot of aggregates in media when the induction was controlled at 0.75% methanol, and two different aggregate forms were found: disulfide bond covalent aggregates and non-covalent aggregates. However, these aggregates almost disappeared when the methanol concentration was controlled at 0.25%, at the same time, cIFN bioactivity of supernatant increased almost 4.48-fold. It is the first time to report the influence of methanol concentration on stability of recombinant protein by P. pastoris. Our results strongly emphasizes the necessity to obtain a maximum of information on the process parameters such as methanol concentration and to maintain strict process control in order to increase the quality of recombinant protein in the high cell-density fermentation of P. pastoris.
%K Pichia pastoris
%K recombinant human consensus interferon
%K methanol concentration
%K aggregates
%K fermentation optimization<br>毕赤酵母
%K 复合α干扰素
%K 甲醇浓度
%K 聚合体
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=042A5D990BB41D614EC7C2A2A3CAE065&yid=9377ED8094509821&vid=DB817633AA4F79B9&iid=59906B3B2830C2C5&sid=2FA47D7E22372F11&eid=7E823D8841E016EC&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=0&reference_num=19