工业微生物
工業微生物
공업미생물
INDUSTRIAL MICROBIOLOGY
2015年
2期
13-20
,共8页
翟成一%徐岩%聂尧%穆晓清
翟成一%徐巖%聶堯%穆曉清
적성일%서암%섭요%목효청
普鲁兰酶%大肠杆菌%质粒稳定性%本底表达%发酵优化
普魯蘭酶%大腸桿菌%質粒穩定性%本底錶達%髮酵優化
보로란매%대장간균%질립은정성%본저표체%발효우화
pullulanase%E. coli%plasmid stability%basal expression%fermentation optimization
通过对产普鲁兰酶的重组大肠杆菌E． coli BL21（DE3）／pET28a-s-pul菌株在发酵过程中质粒稳定性和普鲁兰酶生成量的考察，发现不同宿主对质粒稳定性及酶活性有重要影响。本文利用E． coli BL21（DE3）pLysS菌株为宿主，构建重组菌E． coli BL21（DE3）pLysS／pET28a-s-pul，通过控制外源蛋白的本底表达，提高了重组菌株的质粒稳定性。优化发酵培养基和发酵条件以后，重组菌产普鲁兰酶能力由480 U／mL提高至627 U／mL，增幅为30．6％。研究结果认为，严格控制外源蛋白的本底表达，是改善重组菌稳定性的重要方法之一。
通過對產普魯蘭酶的重組大腸桿菌E． coli BL21（DE3）／pET28a-s-pul菌株在髮酵過程中質粒穩定性和普魯蘭酶生成量的攷察，髮現不同宿主對質粒穩定性及酶活性有重要影響。本文利用E． coli BL21（DE3）pLysS菌株為宿主，構建重組菌E． coli BL21（DE3）pLysS／pET28a-s-pul，通過控製外源蛋白的本底錶達，提高瞭重組菌株的質粒穩定性。優化髮酵培養基和髮酵條件以後，重組菌產普魯蘭酶能力由480 U／mL提高至627 U／mL，增幅為30．6％。研究結果認為，嚴格控製外源蛋白的本底錶達，是改善重組菌穩定性的重要方法之一。
통과대산보로란매적중조대장간균E． coli BL21（DE3）／pET28a-s-pul균주재발효과정중질립은정성화보로란매생성량적고찰，발현불동숙주대질립은정성급매활성유중요영향。본문이용E． coli BL21（DE3）pLysS균주위숙주，구건중조균E． coli BL21（DE3）pLysS／pET28a-s-pul，통과공제외원단백적본저표체，제고료중조균주적질립은정성。우화발효배양기화발효조건이후，중조균산보로란매능력유480 U／mL제고지627 U／mL，증폭위30．6％。연구결과인위，엄격공제외원단백적본저표체，시개선중조균은정성적중요방법지일。
It was found that different host bacteria would seriously affect the plasmid stability and pullulanase activity by investigating the recombinant E. coli BL21 (DE3)/pET28a-s-pul producing pullulanase. In this study,a recombinant E. coli BL21(DE3)pLysS/pET28a-s-pul was constructed with a new strain named E. coli BL21(DE3)pLysS,which suc-cessfully controlled the basal expression and improved the plasmid stability. After optimizing the medium and fermentation conditions,the pullulanase activity was enhanced from 480 U/mL to 627 U/mL with an increase of 30. 6%. The results of this research illustrated that strictly controlling the basal expression of the foreign protein was one of the important methods to improve the stability of recombinant bacteria.
