米曲霉沪酿3.042原生质体制备和再生的研究  

Protoplast preparation and regeneration of Aspergillus oryzae 3.042

作　　者：李铁[1] 臧威[1] 刘井权[1] 孙剑秋[1] 肖静[1] 张淑园[1] 张兰兰[1] 

机构地区：[1]齐齐哈尔大学生命科学与工程学院,黑龙江齐齐哈尔161006

出　　处：《中国酿造》2007年第2期19-22,共4页China Brewing

基　　金：黑龙江省教育厅科学技术研究项目(10541259)

摘　　要：研究了酶液组成、酶解温度、酶解时间、渗透压稳定剂、培养基成分、再生培养方式等因素对米曲霉沪酿3.042原生质体制备和再生的影响,建立了米曲霉沪酿3.042原生质体制备和再生的适宜方法,即在查氏液体培养基中28℃培养10h,菌体用0.8mol/L NaCl配制的3%溶壁酶、1%纤维素酶、1%蜗牛酶、0.5%溶菌酶的复合酶液,30℃酶解4h￣6h后,经0.8mol/L NaCl再生固体培养基,双层平板培养法进行原生质体再生,可获得超过70%的再生率。为进一步通过原生质体技术选育优良的酱制品生产用菌株奠定了方法学基础。The factors affecting protoplast formation and regeneration of Aspergillus oryzae 3.042 were studied including composition of enzyme mixture, enzymolysis temperature, enzymolysis time, osmotic pressure stabilizer, medium composition, culture methods and so on. The appropriate method for protoplast formation and regeneration were developed. A. oryzae 3.042 was cultured in Czapek liquid medium for 10 h, then the mycelia were gathered by centrifugation and digested by enzyme mixture consisting of 0.8 mol/L NaCI, 3% lywallzyme, 1% cellulase, 1% snailase and 0.5% lysozyme at 30℃ for 4-6 h, followed by the regeneration on solid medium containing 0.8 mol/L NaCI by double-layers cultivation. A regeneration rate of 70% was achieved. This study provided a new method to screen good strains for sauce production by protoplast technology.

关 键 词：米曲霉沪酿3.042 原生质体 制备 再生 

分 类 号：Q813.1[生物学—生物工程]