Isolation and Characterization of <em>Mucor circinelloides pyrG</em> Negative Mutant Strain

doi:DOI:10.13523/j.cb.20160306

China Biotechnology

2016, Vol. 36

Issue (3): 38-42 DOI: DOI:10.13523/j.cb.20160306

Isolation and Characterization of Mucor circinelloides pyrG Negative Mutant Strain

ZHANG Ying-tong, CHEN Hai-qin, SONG Yuan-da, ZHANG Hao, CHEN Yong-quan, CHEN Wei

State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi 214122, China

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Abstract

To generate genetic selective marker for constructing super-producing lycopene strains, chemical mutagenesis was applied in mutagenizing Mucor circinelloides MU616. Five uracil auxotroph strains of Mucor circcinelloides were isolated with nitrosoguanidine and selected by resistance to 5-fluoroorotic acid, and these mutants were unable to grow in the minimal medium even after 5 days' culture. Plasmid pEPM1 which harbors the pyrG gene encoding orotidine-5'-monophosphate decarboxylase in Mucor circinelloides were used to transformed the representative uracil auxotroph strains, the result demonstrated that mutants Mt1, Mt4 and Mt5 were deficient in pyrG gene. However, the lycopene content analysis shown that only Mt4 maintained the capacity of accumulating lycopene[(1 648±185)μg/g and (3 234±281)μg/g under dark and light conditions respectively], in addition, the biomass[(9.0±0.6)g/L] and growth properties of Mt4 is also similar to the original strain MU616. It suggested that Mt4 could be used as the receipt strain for the subsequent molecular manipulation. The acquisition of the pyrG mutant strain is very important for construction of the super-producing lycopene strains by molecular genetic manipulation.

Key words： Uracil auxotroph Mucor circinelloides Chemical mutagenesis Electrotansformation Lycopene

Received: 13 October 2015 Published: 19 November 2015

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ZHANG Ying-tong, CHEN Hai-qin, SONG Yuan-da, ZHANG Hao, CHEN Yong-quan, CHEN Wei. Isolation and Characterization of Mucor circinelloides pyrG Negative Mutant Strain. China Biotechnology, 2016, 36(3): 38-42.

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https://manu60.magtech.com.cn/biotech/DOI:10.13523/j.cb.20160306 OR https://manu60.magtech.com.cn/biotech/Y2016/V36/I3/38

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