Efficient modification of the Pseudomonas aeruginosa toxin 2-heptyl-1-hydroxyquinolin-4-one by three Bacillus glycosyltransferases with broad substrate ranges.
Efficient modification of the Pseudomonas aeruginosa toxin 2-heptyl-1-hydroxyquinolin-4-one by three Bacillus glycosyltransferases with broad substrate ranges.
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DOI：
10.1016/j.jbiotec.2019.11.015
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发表时间：
2019-11
影响因子：
4.1
通讯作者：
S. Thierbach;P. Sartor;O. Yücel;S. Fetzner
中科院分区：
文献类型：
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作者：
S. Thierbach;P. Sartor;O. Yücel;S. Fetzner
天然和合成产物的糖基化可以改变苷元的物理、化学和药理性质。铜绿假单胞菌产生的一种有效的呼吸抑制剂2-庚基-1-羟基喹啉-4-酮(HQNO)最近在枯草芽孢杆菌168菌株中被证明转化为毒性较低的2-heptyl-1-(β-D-glucopyranosydyl)-quinolin-4-one，。在这项研究中，我们比较了几种芽孢杆菌的基因组。确定HQNO糖基化的候选酶。所有三个(推测的)UDP-糖基转移酶(GT)都是B。供试的3株枯草杆菌YjiC、YdhE和YojK对HQNO具有较强的糖基化能力，其中YjiC对HQNO的转化率(Kcat)最高，为4.6 S−1，YdhE对HQNO的Km值最低，为9.1μM，三种GT均以UDP-葡萄糖为主，而YdhE对TDP-葡萄糖的作用类似。在所测试的配基中，HQNO是所有三种GT的首选底物，但它们也显示出对Thep的活性。铜绿假单胞菌外显子产物绿青素、2-庚基-3-羟基喹啉-4(1H)-酮(假单胞菌喹诺酮信号)和2，4-二羟基喹啉，植物来源的抗菌素香草醛和槲皮素，以及大环内酯类抗生素泰乐菌素A。我们的结果强调了YjiC、YdhE和YojK的混杂和底物灵活性，并暗示了YjiC、YdhE和YojK在天然毒素抗性中的生理作用。枯草杆菌。尤其是YdhE似乎是天然产物糖化工程的一种有吸引力的生物催化剂。
Glycosylation of natural and synthetic products can alter the physical, chemical and pharmacological properties of the aglycon. Conversion of 2-heptyl-1-hydroxyquinolin-4-one (HQNO), a potent respiratory inhibitor produced byPseudomonas aeruginosa, to the less toxic 2-heptyl-1-(β-D-glucopyranosydyl)-quinolin-4-one, was recently demonstrated forBacillus subtilisstrain 168. In this study, we compared the genomes of severalBacillusspp. to identify candidate enzymes for HQNO glucosylation. All three (putative) UDP-glycosyltransferases (GT) ofB. subtilis 168 tested, YjiC, YdhE and YojK, were capable of HQNO glucosylation, with YjiC showing the highest turnover rate (kcat) of 4.6 s−1, and YdhE exhibiting the lowest Kmvalue for HQNO of 9.1 μM. All three GT predominantly utilized UDP-glucose, but YdhE was similarly active with TDP-glucose. Among the aglycons tested, HQNO was the preferred substrate of all three GT, but they also showed activities toward theP. aeruginosaexoproducts pyocyanin, 2-heptyl-3-hydroxyquinolin-4(1H)-one (thePseudomonasquinolone signal) and 2,4-dihydroxyquinoline, the plant derived antimicrobials vanillin and quercetin, and the macrolide antibiotic tylosin A. Our results underline the promiscuity and substrate flexibility of YjiC, YdhE and YojK, and suggest a physiological role in natural toxin resistance ofB. subtilis. Especially YdhE appears to be an attractive biocatalyst for the glycoengineering of natural products.