摘要
在利用重组毕赤酵母表达瑞替普酶(Reteplase)时,遇到了较为严重的蛋白降解。为了抑制Reteplase的降解,分别研究了pH和氨基酸、蛋白胨等添加物对降解的影响。结果表明,摇瓶培养时用氨水调节pH,并将pH恒定在6.5能有效地抑制降解,这一结果在5L罐上也得到了验证,表达量提高到487mg/L。添加0.1%的Leu或Ala和将蛋白胨用量增加至6%时,Reteplase的降解也能得到有效的控制。
Severe degradation of reteplase expressed by Pichia pastoris was observed. Effects of pH and addition of amino acids and polypeptone were investigated in order to minimize the degradation of target protein. In shake flasks, it was suggested that rPA degradation could be alleviated effectively at pH 6.5 when ammonia was used to adjust the pH. When the results were applied to 5L fermentor, the concentration of rPA was increased to 487mg/L. At the same time, the addition of 0.1% Leu or Ala and the increasement of polypeptone content to 6 % in the medium could also reduce rPA degradation to some extent.
出处
《工业微生物》
CAS
CSCD
北大核心
2006年第2期7-11,共5页
Industrial Microbiology
基金
国家高技术研究发展计划"863计划"(No.2002AA217021
2002AA2Z3451)
作者简介
邹敏（1979～），女，硕士研究生。
