Protoplast preparation and reversion to the normal filamentous growth in antibiotic-producing uncommon actinomycetes
Protoplast preparation and reversion to the normal filamentous growth in antibiotic-producing uncommon actinomycetes
复制标题
DOI：
10.1038/ja.2009.127
复制
发表时间：
2010-02-01
影响因子：
3.3
通讯作者：
Beltrametti, Fabrizio
中科院分区：
文献类型：
--
作者：
Marcone, Giorgia Letizia;Carrano, Lucia;Beltrametti, Fabrizio
原生质体制备、再生和融合是那些研究较少的、不适用于当前分子生物学方案的生物技术上有价值的微生物的基本工具。以玫瑰花平双孢霉（Planobispora rosea）为出发菌株，采用鸡蛋清溶菌酶（HEWL）和球孢链霉菌变溶菌素（Streptomyces globisporus mutanolysin）联合应用的原生质体制备和再生方法，对链孢囊菌科（Streptosporangiaceae）、小单孢菌科（Micromonosporaceae）和链霉菌科（Streptomycetaceae）的一组产芽孢丝状放线菌进行原生质体制备和再生。根据菌株，在消化溶液中孵育几小时至1或2天后，从100 ml培养物中获得10（7）-10（9）个原生质体。原生质体回复到正常丝状生长的效率从0.1%到接近50%不等。对三种代表性菌株（Nonomuraea sp. ATCC 39727、Actinoplanes teichomyceticus ATCC 31121和Streptomyces coelicolor A3（2））的细胞壁肽聚糖的分析证明了聚糖链和肽链中的结构变异，这可能是对细胞消化和原生质体再生处理的不同反应的原因。抗生素杂志（2010）63，83-88; doi：10.1038/ja.2009.127; 2010年1月8日在线发表
Protoplast preparation, regeneration and fusion represent essential tools for those poorly studied biotechnologically valuable microorganisms inapplicable with the current molecular biology protocols. The protoplast production and regeneration method developed for Planobispora rosea and using the combination of hen egg-white lysozyme (HEWL) and Streptomyces globisporus mutanolysin was applied to a set of antibiotic-producing filamentous actinomycetes belonging to the Streptosporangiaceae, Micromonosporaceae and Streptomycetaceae. 10(7)-10(9) protoplasts were obtained from 100 ml of culture, after incubation times in the digestion solution ranging from a few hours to 1 or 2 days depending on the strain. The efficiency of protoplast reversion to the normal filamentous growth varied from 0.1 to nearly 50%. Analysis of cell wall peptidoglycan in three representative strains (Nonomuraea sp. ATCC 39727, Actinoplanes teichomyceticus ATCC 31121 and Streptomyces coelicolor A3(2)) has evidenced structural variations in the glycan strand and in the peptide chain, which may account for the different response to cell digestion and protoplast regeneration treatments. The Journal of Antibiotics (2010) 63, 83-88; doi:10.1038/ja.2009.127; published online 8 January 2010