Site-directed mutagenesis of UbiA to promote menaquinone biosynthesis in Elizabethkingia meningoseptica
Site-directed mutagenesis of UbiA to promote menaquinone biosynthesis in Elizabethkingia meningoseptica
复制标题
UbiA 的定点诱变促进 Elizabethkingia meningoseptica 的甲基萘醌生物合成
DOI：
10.1016/j.procbio.2017.05.002
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发表时间：
2017-07-01
影响因子：
4.4
通讯作者：
Xu, Ping
中科院分区：
文献类型：
--
作者：
Liu, Yan;Ding, Xiu-min;Xu, Ping
为了提高脑膜炎败血症克氏菌的甲萘醌（MK）含量，通过定点突变抑制4-羟基苯甲酸八异戊二烯基转移酶（UbiA）的活性，进而阻断泛醌（UQ）的生物合成途径。对除L174和G211外的14个保守残基进行突变，以分析定点突变的效果。12个突变体中UbiA在mRNA和蛋白水平的表达均降低，导致UQ浓度降低。根据MenA的表达水平，将12个突变体分为两组。第二组如N72 A、D 76 A、K81 A、L139 A和D198 A增强MenA的表达，使MK产量分别增加127.1%、87.9%、96.2%、109.7%和130.0%。一般而言，通过定点突变大肠杆菌UbiA的活性位点来阻断UQ合成途径。脑膜炎败血症是一个很有前途的策略，以提高MK生产在E。脑膜炎
To increase the menaquinone (MK) content of an Elizabethkingia meningoseptica, site-directed mutagenesis was generated to suppress 4-hydroxybenzoate octaprenyl transferase (UbiA) activity and subsequently blocked the ubiquinone (UQ) biosynthesis pathway. Fourteen conserved residues except L174 and G211 were mutated to analyze the effect of site-directed mutagenesis. The expression of UbiA in twelve mutants was decreased in both mRNA and protein levels, which resulted in the decrease of UQ concentration. Based on MenA expression level, 12 mutants were divided into two groups. Second group such as N72A, D76A, K81A, L139A, and D198A enhanced the expression of MenA, which increased MK production by 127.1%, 87.9%, 96.2%, 109.7% and 130.0% in wt-EmUbiA, respectively. In general, blocking UQ synthesis pathway for by site-directed mutagenesis of the active site of UbiA in E. meningoseptica was a promising strategy to increase MK production in E. meningoseptica.