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首页> 外文期刊>Bioprocess and Biosystems Engineering >Ethambutol-mediated cell wall modification in recombinant Corynebactetium glutamicum increases the biotransformation rates of cyclohexanone derivatives
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Ethambutol-mediated cell wall modification in recombinant Corynebactetium glutamicum increases the biotransformation rates of cyclohexanone derivatives

机译：乙草胺介导的重组谷氨酸棒状杆菌细胞壁修饰可提高环己酮衍生物的生物转化率

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摘要

The effects of structural modification of cell wall on the biotransformation capability by recombinant Corynebacterium glutamicum cells, expressing the chnB gene encoding cyclohexanone monooxygenase of Acine-tobacter calcoaceticus NCIMB 9871, were investigated. Baeyer-Villiger oxygenation of 2-(2'-acetoxyethyl) cyclohexanone (MW 170 Da) into R-7-(2'-acetoxyethyl)-2-oxepanone was used as a model reaction. The whole-cell biotransformation followed Michaelis-Menten kinetics. The V_(max) and K_s values were estimated as 96.8 Ug~(-1) of dry cells and 0.98 mM, respectively. The V_(max) was comparable with that of cyclohexanone oxygenation, whereas the K_s was almost eightfold higher. The K_s value of 2-(2'-acetoxyethyl) cyclohexanone oxygenation was reduced by ca. 30% via altering the cell envelop structure of C. glutamicum with ethambutol, which inhibits arabi-nosyl transferases involved in the biosynthesis of cell wall arabinogalactan and mycolate layers. The higher whole-cell biotransformation rate was also observed in the oxygenation of ethyl 2-cyclohexanone acetate upon ethambutol treatment of the recombinant C. glutamicum. Therefore, it was assumed that the biotransformation efficiency of C. glutamicum-based biocatalysts, with respect to medium- to large-sized lipophilic organic substrates (MW > ca. 170), can be enhanced by engineering their cell wall outer layers, which are known to function as a formidable barrier to lipophilic molecules.
机译：研究了细胞壁的结构修饰对重组谷氨酸棒状杆菌细胞的生物转化能力的影响，所述重组谷氨酸棒状杆菌细胞表达编码不动杆菌-钙钙乙酸NCIMB 9871的环己酮单加氧酶的chnB基因。将2-（2'-乙酰氧基乙基）环己酮（MW 170 Da）的Baeyer-Villiger氧化成R-7-（2'-乙酰氧基乙基）-2-氧戊酮用作模型反应。全细胞生物转化遵循Michaelis-Menten动力学。 V_（max）和K_s值分别估计为96.8 Ug〜（-1）干细胞和0.98 mM。 V_（max）与环己酮氧化相当，而K_s几乎高出八倍。 2-（2'-乙酰氧基乙基）环己酮氧化的K_s值降低了约30％是通过用乙胺丁醇改变谷氨酸棒状杆菌的细胞包膜结构来实现的，乙胺丁醇抑制与细胞壁阿拉伯半乳聚糖和霉菌酸酯层的生物合成有关的阿拉伯糖苷基转移酶。在乙胺丁醇处理重组谷氨酸棒状杆菌后，在2-环己酮乙基乙酸乙酯的氧合作用中也观察到更高的全细胞生物转化率。因此，据推测，相对于中型至大型亲脂性有机底物（MW>约170），谷氨酸棒杆菌基生物催化剂的生物转化效率可以通过对其细胞壁外层进行改造来提高。已知对亲脂性分子具有强大的屏障作用。

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