Protoplasting Improvement of Industrial Citrate-Producing Aspergillus niger for DNA Transformation

doi:10.3979//j.issn.1673-1689.2016.09.010

Home > Archive>Volume 35, Issue 9, 2016 >963-970. DOI:10.3979//j.issn.1673-1689.2016.09.010

Protoplasting Improvement of Industrial Citrate-Producing Aspergillus niger for DNA Transformation
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                        10.3979//j.issn.1673-1689.2016.09.010
                    
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This article aimed to optimize protoplast formation conditions of industrial citrate-producing Aspergillus niger to establish genetic transformation system. As protoplasts were formed by enzyme digestion，components of enzyme mixtures，digestion conditions and hyphae growth conditions were studied. The optimized enzyme mixture contained 5 mg/mL lysing enzyme，0.2 U/mL chitinase and 460 U/mL glucuronidase. The best conditions for protoplasting were using 0.7 M KCl as osmotic stabilizer，thickness of pellets hyphae for 50 ?滋m，cell weight of 0.003 g/mL. Furthermore，addition of Mn2+ in culture medium helped reduce the chitinase concentration necessary for protoplasting. In addition，glucuronidase in culture medium facilitated conidia separation during germination，further improved protoplast formation. Finally，two cassettes were inserted into genome simultaneously.

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YIN Xian, PANG Dongqia, HAN Ruizhi, LI Jianghua, LIU Long, DU Guocheng, CHEN Jian. Protoplasting Improvement of Industrial Citrate-Producing Aspergillus niger for DNA Transformation[J]. Journal of Food Science and Biotechnology,2016,35(9):963-970.

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Online: November 01,2016
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