Xiaojing Wang1,2,3, Kai Wu2,3, Hanzhi Zhang4, Jing Liu2,5, Zhijun Yang2,3, Jing Bai6, Hao Liu7, Lei Shao8,9. 1. Affiliated Zhoupu Hospital, Shanghai University of Medicine and Health Sciences, Shanghai, China. 2. Microbial Pharmacology Laboratory, Shanghai University of Medicine and Health Sciences, Shanghai, China. 3. School of Pharmacy, Shanghai University of Medicine and Health Sciences, Shanghai, China. 4. Abiochem Biotechnology Co., Ltd, Shanghai, China. 5. School of Medical Technology, Shanghai University of Medicine and Health Sciences, Shanghai, China. 6. School of Chemistry and Life Sciences, Suzhou University of Science and Technology, Suzhou, China. 7. Department of Antibiotics and Microorganisms, Shanghai Institute for Food and Drug Control, Shanghai, China. liuhao1968@hotmail.com. 8. Microbial Pharmacology Laboratory, Shanghai University of Medicine and Health Sciences, Shanghai, China. shaolei00@gmail.com. 9. School of Pharmacy, Shanghai University of Medicine and Health Sciences, Shanghai, China. shaolei00@gmail.com.
Abstract
OBJECTIVES: Polymyxins are antibacterial polypeptides used as "last resort" therapy option for multidrug-resistant Gram-negative bacteria. The expansion of polymyxin-resistant infections has inspired development of novel polymyxin derivatives, and deacylation is one of the critical steps in generating those antibiotics. Deacylase from Actinoplanes utahensis hydrolyze the acyl moieties of echinocandins, and also efficiently deacylates daptomycin, ramoplanin and other important antibiotics. Here, deacylase was studied considering its potential usefulness in deacylating polymyxin B1. RESULTS: All the six recombinant strains containing the deacylase gene catalyzed hydrolysis of polymyxin B1, yielding cyclic heptapeptides. The efficiency of recombinant S. albus (SAL701) was higher than that of the others, and deacylation was the most efficient at 40 °C in 0.2 M Tris buffer (pH 8.0) with 0.2 M Mg2+. The optimal substrate concentration of SAL701 was increased from 2.0 to 6.0 g/L. SAL701 was highly thermostable, showing no loss of activity at 50 °C for 12 h, and the mycelia could be recycled at least three times without loss of catalytic activity. SAL701 could not deacylate β-lactam substrate such as penicillin G and cephalosporin C. Deacylase catalyzes the amide bond 1 closest to the nucleus of polymyxin B1 rather than the other bond, suggesting that it has high catalytic site specificity. Homology modeling and the docking results implied that Thr190 in deacylase could facilitate hydrolysis with high regioselectivity. CONCLUSIONS: These results show that SAL701 is effective in increasing the cyclic heptapeptide moiety of polymyxin B1. These properties of the biocatalyst may enable its development in the industrial production of polymyxins antibiotics.
OBJECTIVES: Polymyxins are antibacterial polypeptides used as "last resort" therapy option for multidrug-resistant Gram-negative bacteria. The expansion of polymyxin-resistant infections has inspired development of novel polymyxin derivatives, and deacylation is one of the critical steps in generating those antibiotics. Deacylase from Actinoplanes utahensis hydrolyze the acyl moieties of echinocandins, and also efficiently deacylates daptomycin, ramoplanin and other important antibiotics. Here, deacylase was studied considering its potential usefulness in deacylating polymyxin B1. RESULTS: All the six recombinant strains containing the deacylase gene catalyzed hydrolysis of polymyxin B1, yielding cyclic heptapeptides. The efficiency of recombinant S. albus (SAL701) was higher than that of the others, and deacylation was the most efficient at 40 °C in 0.2 M Tris buffer (pH 8.0) with 0.2 M Mg2+. The optimal substrate concentration of SAL701 was increased from 2.0 to 6.0 g/L. SAL701 was highly thermostable, showing no loss of activity at 50 °C for 12 h, and the mycelia could be recycled at least three times without loss of catalytic activity. SAL701 could not deacylate β-lactam substrate such as penicillin G and cephalosporin C. Deacylase catalyzes the amide bond 1 closest to the nucleus of polymyxin B1 rather than the other bond, suggesting that it has high catalytic site specificity. Homology modeling and the docking results implied that Thr190 in deacylase could facilitate hydrolysis with high regioselectivity. CONCLUSIONS: These results show that SAL701 is effective in increasing the cyclic heptapeptide moiety of polymyxin B1. These properties of the biocatalyst may enable its development in the industrial production of polymyxins antibiotics.
Authors: M Debono; B J Abbott; J R Turner; L C Howard; R S Gordee; A S Hunt; M Barnhart; R M Molloy; K E Willard; D Fukuda Journal: Ann N Y Acad Sci Date: 1988 Impact factor: 5.691
Authors: Jie He; Kamilia Abdelraouf; Kimberly R Ledesma; Diana S-L Chow; Vincent H Tam Journal: Int J Antimicrob Agents Date: 2013-08-22 Impact factor: 5.283
Authors: Predrag Cudic; Douglas C Behenna; James K Kranz; Ryan G Kruger; A Joshua Wand; Yuri I Veklich; John W Weisel; Dewey G McCafferty Journal: Chem Biol Date: 2002-08
Authors: Romeo Ciabatti; Sonia I Maffioli; Gianbattista Panzone; Augusto Canavesi; Elena Michelucci; Paolo S Tiseni; Ettore Marzorati; Anna Checchia; Matteo Giannone; Daniela Jabes; Gabriella Romano; Cristina Brunati; Gianpaolo Candiani; Franca Castiglione Journal: J Med Chem Date: 2007-06-02 Impact factor: 7.446