文献类型：期刊文献

中文题名：顶头孢霉菌原生质体的制备和再生条件的优化

英文题名：Optimization of Protoplast Preparation and Regeneration Conditions of Acremonium chrysogenum

作者：杨颂[1];赵春云[1];欧阳立明[1];储炬[1];张嗣良[1]

机构：[1]华东理工大学生物反应器国家重点实验室,上海200237

年份：2016

卷号：47

期号：6

起止页码：687

中文期刊名：中国医药工业杂志

外文期刊名：Chinese Journal of Pharmaceuticals

收录：CSTPCD;;北大核心:【北大核心2014】；CSCD:【CSCD2015_2016】；

基金：国家国际科技合作专项项目(2013DFG32630);国家"973"重点基础研究发展计划(012CB721006);国家重大新药创制科技重大专项(2011ZX09203-001-03)

语种：中文

中文关键词：顶头孢霉;原生质体;再生

外文关键词：Acremonium chrysogenum; protoplast; regeneration

摘要：考察了细胞壁降解酶的种类、酶解时间、培养基种类和涂布方式对顶头孢霉原生质体制备和再生的影响,结果提示顶头孢霉菌丝形态对原生质的制备和再生也有重要影响。顶头孢霉生长过程可分为3个时期：细长时期、中度膨大时期和高度膨大时期。本试验利用显微图像分析法确定了各时期菌丝的形态参数,描述了培养过程中的菌丝形态变化规律,以及不同菌丝形态对原生质体释放和再生率的影响。最终优化条件为：选择中度膨大初期的菌丝体,使用1%溶壁酶,30℃酶解2 h后采用双层平板法接种于原生质体再生培养基I（HCM）中。每1 ml原生质体悬浮液中原生质体的释放数可达2.51×10^8个,再生率大于20%。
The influences of cell wall degrading enzymes, enzymolysis time, composition of culture media and spreading ways on protoplast preparation and regeneration of Acremonium chrysogenum were systematically examined. The morphology of hyphae was found to have significant impact on the quality of protoplast as well. The morphologies of Acremonium chrysogenum could be described as elongated hyphae, moderately swollen hyphae and highly swollen hyphae. The morphology parameters and the evolution of mycelia morphology during culture were quantitatively described by microscopic image analysis. The relationship between hyphae morphology and the protoplast quantity were studied. The optimum conditions of protoplast preparation were as follows： moderately swollen hyphae of Acremonium chrysogenum was digested by 1% lysing enzyme for 2 h at 30 ℃, and then the protoplasts were cultured on HCM medium with double-plated inoculation method. The number of protoplast in the broth reached 2.51 × 10^8 per milliliter, and the protoplast regeneration ratio was more than 20 %.