Li-Qun Jin1,2,3, Zhe-Wen Xu1,2,3, Xiao-Hui Men1,2,3, Zhi-Qiang Liu4,5,6, Yu-Guo Zheng1,2,3. 1. Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, 310014, People's Republic of China. 2. Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, Hangzhou, 310014, People's Republic of China. 3. The National and Local Joint Engineering Research Center for Biomanufacturing of Chiral Chemicals, Zhejiang University of Technology, Hangzhou, 310014, People's Republic of China. 4. Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, 310014, People's Republic of China. microliu@zjut.edu.cn. 5. Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, Hangzhou, 310014, People's Republic of China. microliu@zjut.edu.cn. 6. The National and Local Joint Engineering Research Center for Biomanufacturing of Chiral Chemicals, Zhejiang University of Technology, Hangzhou, 310014, People's Republic of China. microliu@zjut.edu.cn.
Abstract
OBJECTIVE: To explore the optimal methods for the protoplast preparation and regeneration of Hirsutella sinensis by optimizing the limiting factors. RESULTS: During the treatment of enzymatic protoplast preparation, mycelium cultured for 7 days was the optimal start material. The maximum protoplast preparation rate of 4.3 × 107 protoplasts/g fresh weight (FW) was obtained after 0.5 h treatment of 1 mg/ml mixed lytic enzymes in KH2PO4-K2HPO4 buffer (pH 5.5) with 0.6 M KCl at 18 °C. As for the protoplast regeneration, the maximum protoplast regeneration rate reached 12.32% through 5 × 103 protoplasts mL-1 cultivated for 20 days in the regeneration medium with 0.6 M mannitol and 1.5% agar. CONCLUSIONS: The preparation and regeneration of H. sinensis protoplasts was firstly established based on process optimization and it provided a foundation for the study of H. sinensis mutagenesis.
OBJECTIVE: To explore the optimal methods for the protoplast preparation and regeneration of Hirsutella sinensis by optimizing the limiting factors. RESULTS: During the treatment of enzymatic protoplast preparation, mycelium cultured for 7 days was the optimal start material. The maximum protoplast preparation rate of 4.3 × 107 protoplasts/g fresh weight (FW) was obtained after 0.5 h treatment of 1 mg/ml mixed lytic enzymes in KH2PO4-K2HPO4 buffer (pH 5.5) with 0.6 M KCl at 18 °C. As for the protoplast regeneration, the maximum protoplast regeneration rate reached 12.32% through 5 × 103 protoplasts mL-1 cultivated for 20 days in the regeneration medium with 0.6 M mannitol and 1.5% agar. CONCLUSIONS: The preparation and regeneration of H. sinensis protoplasts was firstly established based on process optimization and it provided a foundation for the study of H. sinensis mutagenesis.
Authors: Shin Yee Fung; Peter Chiew Hing Cheong; Nget Hong Tan; Szu Ting Ng; Chon Seng Tan Journal: Int J Med Mushrooms Date: 2018 Impact factor: 1.921
Authors: Jan Martel; Yun-Fei Ko; Jian-Ching Liau; Chien-Sheng Lee; David M Ojcius; Hsin-Chih Lai; John D Young Journal: Trends Biotechnol Date: 2017-11 Impact factor: 19.536