%0 Journal Article
%T Expression of Vitreoscilla hemoglobin improves recombinant lipase production in Pichia pastoris<br>毕赤酵母中表达透明颤菌血红蛋白提高重组脂肪酶的表达
%A Xiaofeng Wang
%A Yongchuan Sun
%A Xuguang Shen
%A Feng Ke
%A Li Xu
%A Yun Liu
%A Yunjun Yan
%A <br>汪小锋
%A 孙永川
%A 申旭光
%A 柯锋
%A 徐莉
%A 刘云
%A 闫云君
%J 生物工程学报
%D 2011
%I 
%X Yarrowia lipolytica lipase Lip2 (YlLip2) is an important industrial enzyme with many potential applications. To alleviate the dissolved oxygen (DO) limitation and improve YlLip2 production during high-cell density fermentation, the YlLip2 gene lip2 and Vitreoscilla hemoglobin (VHb) gene vgb were co-expressed in Pichia pastoris under the control of AOX1 and PsADH2 promoter, respectively. The PsADH2 promoter from Pichia stipitis could be activated under oxygen limitation. The SDS-PAGE and CO-difference spectrum analysis indicated that VHb and YlLip2 had successfully co-expressed in recombinant strains. Compared with the control cells (VHb?, GS115/9Klip2), the expression levels of YlLip2 in VHb-expressing cells (VHb+, GS115/9Klip2-pZPVT) under oxygen limitation were improved 25% in shake-flask culture and 83% in a 10 L fermentor. Moreover, the VHb+ cells displayed higher biomass than VHb? cells at lower DO levels in a 10 L fermentor. In this study, we also achieved a VHb-expressing clone harboring multicopy lip2 gene (GS115/9Klip2-pZPVTlip2 49#), which showed the maximum lipolytic activity of 33 900 U/mL in a 10 L fermentor under lower DO conditions. Therefore, it can be seen that expression of VHb with PsADH2 promoter in P. pastoris combined with increasing copies of lip2 gene is an effective strategy to improve YlLip2 production.
%K lipase
%K Vitreoscilla hemoglobin
%K PsADH2 promoter
%K Yarrowia lipolytica
%K Pichia pastoris
%K expression<br>脂肪酶，透明颤菌血红蛋白，PsADH2启动子，解脂耶氏酵母，毕赤酵母，表达
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=042A5D990BB41D615DA19DA6CFEE5A00&yid=9377ED8094509821&vid=DB817633AA4F79B9&iid=59906B3B2830C2C5&sid=290C357E8EC0A94B&eid=231F9A307C169827&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=0&reference_num=16