旨在研究 EDTA 预处理、菌龄、酶解浓度、酶解温度及酶解时间对 Sphingomonas sp. ATCC 31555原生质体制备与再生的影响。结果表明，菌龄10 h，12 mmol/L EDTA 预处理，溶菌酶浓度45μg/mL、酶解温度28℃、酶解时间25 min。得到原生质体的形成率达97.3%，再生率达33.9%。研究结果为菌株进行原生质体融合、基因组改组等选育优良菌种提供参考。%The effects of EDTA pretreatment,cell age and the concentration,temperature and work time of enzymolysis on protoplast formation and regeneration by Sphingomonas sp. ATCC 31555 were studied. The results showed that the formation rate and regeneration rate reached 97.3% and 33.9% respectively,when the cells cultivated 10 h,pretreated by 12 mmol/L EDTA,using 45 μg/mL of lysozyme, enzymolysis at 28℃ for 25 min. The result of this paper provides a reference for breeding fine microbial strains by protoplast fusion and genome shuffling.
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