Wang L F, Wong S L, Lee S G, Kalyan N K, Hung P P, Hilliker S, Doi R H
Department of Biochemistry and Biophysics, University of California, Davis 95616.
Gene. 1988 Sep 15;69(1):39-47. doi: 10.1016/0378-1119(88)90376-9.
Using the signal peptide of the Bacillus subtilis subtilisin gene (aprE) and a synthetic cDNA corresponding to the mature region of the human atrial natriuretic alpha-factor (hANF), we have constructed a secretion vector. B. subtilis cells, when transformed with this vector, secrete immunoreactive hANF peptides into the medium at about 500 micrograms/liter. The hANF is the first human gene product to be secreted from B. subtilis using this signal peptide. We have used promoters active during vegetative growth or sporulation and hosts deficient in several extracellular proteases but some proteolysis of the secretion products still occurs. In addition, both cell growth and sporulation are adversely affected by hANF production. Possible explanations for this observation are inefficient secretion of the atrial hormone or toxicity of the precursor or mature peptide.
利用枯草芽孢杆菌枯草杆菌蛋白酶基因（aprE）的信号肽和对应于人心房利钠α因子（hANF）成熟区的合成cDNA，我们构建了一个分泌载体。用该载体转化枯草芽孢杆菌细胞后，能以约500微克/升的量将免疫反应性hANF肽分泌到培养基中。hANF是首个利用该信号肽从枯草芽孢杆菌中分泌的人类基因产物。我们使用了在营养生长或孢子形成过程中活跃的启动子以及几种细胞外蛋白酶缺陷的宿主，但分泌产物仍会发生一些蛋白水解。此外，hANF的产生对细胞生长和孢子形成均有不利影响。对此观察结果的可能解释是心房激素分泌效率低下或前体或成熟肽具有毒性。
