Ebisu S, Mori M, Takagi H, Kadowaki K, Yamagata H, Tsukagoshi N, Udaka S
Research Laboratory, Higeta Shoyu Co., Ltd., Chiba, Japan.
J Ind Microbiol. 1993 Feb;11(2):83-8. doi: 10.1007/BF01583679.
An expression-secretion vector, pMK300, was constructed to express the Aspergillus oryzae Taka-amylase A (Taa) cDNA. The promoter and signal peptide regions of the HWP (a major cell wall protein of Bacillus brevis HPD31) gene on pMK300 were efficiently utilized in B. brevis HPD31 and a large amount of Taa (22 mg/l) was secreted into the medium. The HWP signal peptide utilized for secretion of Taa was correctly processed during the protein transport across the membrane. The enzymatic properties of Taa produced by B. brevis HPD31 were the same as those of the Aspergillus oryzae Taa in several respects; specific activity, thermal and pH stabilities, and temperature and pH optima. These results, in combination with previous results, indicate that B. brevis HPD31 could be used to produce extracellularly foreign proteins of diverse origins as functional proteins.
构建了一种表达分泌载体pMK300，用于表达米曲霉Taka淀粉酶A（Taa）的cDNA。pMK300上的HWP（短短芽孢杆菌HPD31的一种主要细胞壁蛋白）基因的启动子和信号肽区域在短短芽孢杆菌HPD31中得到有效利用，大量的Taa（22mg/L）分泌到培养基中。用于Taa分泌的HWP信号肽在蛋白质跨膜转运过程中被正确加工。短短芽孢杆菌HPD31产生的Taa的酶学性质在几个方面与米曲霉Taa相同；比活性、热稳定性和pH稳定性以及温度和pH最佳值。这些结果与先前的结果相结合，表明短短芽孢杆菌HPD31可用于细胞外产生多种来源的外源蛋白作为功能蛋白。
