Borisova Svetlana A, Circello Benjamin T, Zhang Jun Kai, van der Donk Wilfred A, Metcalf William W
Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.
Chem Biol. 2010 Jan 29;17(1):28-37. doi: 10.1016/j.chembiol.2009.11.017.
Rhizocticins are phosphonate oligopeptide antibiotics containing the C-terminal nonproteinogenic amino acid (Z)-l-2-amino-5-phosphono-3-pentenoic acid (APPA). Here we report the identification and characterization of the rhizocticin biosynthetic gene cluster (rhi) in Bacillus subtilis ATCC6633. Rhizocticin B was heterologously produced in the nonproducer strain Bacillus subtilis 168. A biosynthetic pathway is proposed on the basis of bioinformatics analysis of the rhi genes. One of the steps during the biosynthesis of APPA is an unusual aldol reaction between phosphonoacetaldehyde and oxaloacetate catalyzed by an aldolase homolog RhiG. Recombinant RhiG was prepared, and the product of an in vitro enzymatic conversion was characterized. Access to this intermediate allows for biochemical characterization of subsequent steps in the pathway.
根瘤菌素是一类膦酸酯寡肽抗生素，其C末端含有非蛋白质ogenic氨基酸(Z)-l-2-氨基-5-膦酰基-3-戊烯酸(APPA)。在此，我们报道了枯草芽孢杆菌ATCC6633中根瘤菌素生物合成基因簇(rhi)的鉴定和表征。根瘤菌素B在非产生菌枯草芽孢杆菌168中异源产生。基于rhi基因的生物信息学分析，提出了一条生物合成途径。APPA生物合成过程中的一个步骤是由醛缩酶同源物RhiG催化的膦酰乙醛和草酰乙酸之间的异常醛醇反应。制备了重组RhiG，并对体外酶促转化产物进行了表征。获得该中间体有助于对该途径后续步骤进行生化表征。
