%0 Journal Article
%T Compared D-amino Acid Oxidase Expression in Different Pichia pastoris Host Strains<br>D-氨基酸氧化酶在不同毕赤酵母宿主菌中的表达比较
%A FENG Mei_Qing
%A HUANG Hai SHI Xun_Long YU Zhi_Liang YUAN Zhong Yi ZHOU Pei
%A <br>冯美卿
%A 黄海
%A 史训龙
%A 余志良
%A 袁中一
%A 周珮
%J 生物工程学报
%D 2004
%I 
%X To compare the DAAO expression level in different Pichia pastoris host strains, the gene encoding DAAO from Trigonopsis variabilis was cloned into plasmid pPIC3.5k and then transformed into P. pastoris GS115 and KM71 respectively. The positive transformants PDK13 (MutS) and PD27 (Mut+) were obtained by PCR analysis. Their optimal and different expression conditions were investigated. To compare with PD27, PDK13 was determined to poss a slower consumption of methanol, a longer induction time, a lower oxygen request and apparently higher expression of DAAO. The highest expression levels were reached up to 2700, 2500 IU/L in shaking flask and 10140, 8463.5 IU/L in fermentor respectively. The over-expression of DAAO can meet its large demand for production of 7-ACA, alpha-keto acid and L-amino acid. In addition, the phenylpyruvate and L-phenylalanine were obtained by crude DAAO reacting with DL-phenylalanine at 37 degrees C for 3h.
%K D
%K amino acid oxidation
%K Pichia pastoris
%K different host strain
%K expression
%K bioconversion
%K DL-phenylalanine<br>D-氨基酸氧化酶
%K 甲醇酵母
%K 不同宿主菌
%K 表达比较
%K DL-苯丙氨酸
%K 拆分
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=A53BC2A2AA369D95&yid=D0E58B75BFD8E51C&vid=A04140E723CB732E&iid=E158A972A605785F&sid=6A9657F54F754BF6&eid=52B9DFFFCC2EB041&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=2&reference_num=10