Genome shuffling of Streptomyces viridochromogenes for improved production of avilamycin
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BIOTECHNOLOGICAL PRODUCTS AND PROCESS ENGINEERING
Genome shuffling of Streptomyces viridochromogenes for improved production of avilamycin Xing-An Lv & Ying-Yan Jin & Yu-Dong Li & Hong Zhang & Xin-Le Liang
Received: 4 May 2012 / Revised: 21 June 2012 / Accepted: 18 July 2012 / Published online: 22 August 2012 # Springer-Verlag 2012
Abstract Avilamycin is one of EU-approved antimicrobial agents in feed industry to inhibit the growth of multidrug-resistant Gram-positive bacteria. Here, we applied a process of combining ribosome engineering and genome shuffling to achieve rapid improvement of avilamycin production in Streptomyces viridochromogenes AS 4.126. The starting mutant population was generated by 60Co γ-irradiation treatments of the spores. After five rounds of protoplast fusion with streptomycin-resistance screening, an improved recombinant E-219 was obtained and its yield of avilamycin reached 1.4 g/L, which was increased by 4.85-fold and 36.8-fold in comparison with that of the shuffling starter Co γ-316 and the ancestor AS 4.126. Furthermore, the mechanism for the improvement of shuffled strains was investigated. Recombinants with enhanced streptomycin resistance exhibited significantly higher avilamycin production and product resistance, probably due to the mutations in the ribosome protein S12. The morphological difference between the parent mutant and shuffled recombinant was observed in conidiospore, and hyphae pellets. The presence of genetic diversity among shuffled populations with varied avilamycin productivity was confirmed by randomly amplified polymorphic DNA analysis. In summary, our results demonstrated that genome shuffling combined with ribosome engineering was a powerful approach for molecular breeding of high-yield industrial strains.
Electronic supplementary material The online version of this article (doi:10.1007/s00253-012-4322-7) contains supplementary material, which is available to authorized users. X.-A. Lv : Y.-Y. Jin : Y.-D. Li : H. Zhang : X.-L. Liang (*) Department of Bioengineering, School of Food Science and Biotechnology, Zhejiang Gongshang University, Hangzhou 310025, China e-mail: [email protected]
Keywords Avilamycin . Genome shuffling . Ribosome engineering . RAPD . Streptomyces viridochromogenes
Introduction Avilamycin, produced by Streptomyces viridochromogenes, is a member of the orthosomycin class of antibiotics and has been approved by EU in animal feed (Buzzetti et al. 1968). Avilamycin can inhibit the growth of multidrug-resistant Gram-positive bacteria by binding exclusively to the 50S ribosomal subunit to inhibit bacterial protein synthesis (Wright 1979; Aarestrup and Jensen 2000). Like the compound everninomicin (also called SCH27899 or ziracin), another member of orthosomycin class, avilamycin may provide an alternative to vancomycin for the treatment of many human infectious diseases (Foster and Rybak 1999). It is an effective antimicrobiotic agent against Staphylococcus pneumoniae, Streptococcus pyogenes, Enterococcus faecalis, Enterococcus faecium,
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