Abstract:

Aflatoxin-detoxifizyme （ADTZ） , being from Armillariella tabescens, can effectively decompose aflatoxins. The 5＇coding region of its eDNA was optimized according to the codon bias of Pichia pastoris to make it secretively express in Pichia pastoris with higher performance. Two-step DNA synthsis was used to synthesize the eDNA sequence being optimized of ADTZ （OPT-ADTZ）. OPT-ADTZ was connected with constitutive plasmid pGAPZaA to construct the reecombinant plasmid pNOA. pNOA was linearized and then transformed into Pichia pastoris GS115. Then codonoptimized ADTZ was constitutively and secretively expressed in Pichia pastoris.

Key words: ADTZ , two-step DNA synthesis, constitutive and secretive expression

摘要：

黄曲霉毒素解毒酶（ADTZ）来源于真菌Armillariellatabescens，它能够有效地分解黄曲霉毒素。为了使重组蛋白rADTZ能在毕氏酵母中高效分泌表达，根据毕氏酵母密码子的偏好性对rADTZ的5’末端的编码区域进行了优化，利用two—stepDNAsynthsis技术合成出ADTZ优化的基因序列OPT—ADTZ，并与组成型表达载体pGAPZaA连接，构建重组质粒pNOA，线性化pNOA后转化至毕氏酵母GS115中，实现了密码子优化的rADTZ组成型分泌表达。

关键词: ADTZ two—step DNA synthesis 组成型分泌表达