环状芽孢杆菌内切-β-1,3-1,4-葡聚糖酶基因（bglBC1）在枯草芽孢杆菌和巨大芽孢杆菌中的过量产生与分泌。
Overproduction and secretion of Bacillus circulans endo-beta-1,3-1,4-glucanase gene (bglBC1) in B. subtilis and B. megaterium.
作者信息
Kim Ji-Yeon
机构信息
Genome Research Center, Inje University, Gimhae 621-749, Korea.
出版信息
Biotechnol Lett. 2003 Sep;25(17):1445-9. doi: 10.1023/a:1025059713425.
A gene coding for endo-beta-1,3-1,4-glucanase (lichenase) containing a recombinant plasmid, pLL200K, was transferred from Bacillus circulans into a new shuttle plasmid, pLLS920, by ligating linearized DNAs of pLL200K and pUB110. B. subtilis RM125 and B. megaterium ATCC14945 transformed with pLLS920 produced the endo-beta-1,3-1,4-glucanase. The enzyme was produced during active growth with maximum activity. The B. subtilis (pLLS920) enzyme was 83 times (8522 mU ml(-1)) more active than that of the gene donor cells (103 mU ml(-1)). The B. megaterium (pLLS920) enzyme was 7 times (735 mU ml(-1)) more active than that of the gene donor cells. While E. coli secreted only about 10% of the produced enzyme, B. subtilis excreted the enzyme completely into the medium and B. megaterium by about 98%. The plasmid pLLS920 was stable in B. megaterium (98%), and in B. subtilis (51%) but not in E. coli (29%).
通过连接pLL200K和pUB110的线性化DNA，将编码含有重组质粒pLL200K的内切-β-1,3-1,4-葡聚糖酶（地衣聚糖酶）的基因从环状芽孢杆菌转移到新的穿梭质粒pLLS920中。用pLLS920转化的枯草芽孢杆菌RM125和巨大芽孢杆菌ATCC14945产生了内切-β-1,3-1,4-葡聚糖酶。该酶在活跃生长期间产生，活性最高。枯草芽孢杆菌（pLLS920）产生的酶活性比基因供体细胞（103 mU ml⁻¹）高83倍（8522 mU ml⁻¹）。巨大芽孢杆菌（pLLS920）产生的酶活性比基因供体细胞高7倍（735 mU ml⁻¹）。虽然大肠杆菌仅分泌约10%产生的酶，但枯草芽孢杆菌将酶完全分泌到培养基中，巨大芽孢杆菌则约98%分泌到培养基中。质粒pLLS920在巨大芽孢杆菌（98%）和枯草芽孢杆菌（51%）中稳定，但在大肠杆菌（29%）中不稳定。
Zotero 插件