常压室温等离子体诱变选育DHA高产菌株
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摘要
以野生型裂殖壶菌(Schizochytrium limacinum SR21)为出发菌株,经过常压室温等离子体(ARTP)诱变,碘乙酸、丙二酸及丙二酸-碘乙酸复合平板筛选,摇瓶发酵及GC定量分析选育DHA高产菌株。结果表明:在功率100 W、气流量10 L/min下,经ARTP诱变15 s,以1. 2 g/L丙二酸作为筛选平板,获得一株诱变菌NA2。摇瓶发酵120 h后,其生物量、油脂总量、DHA产量与原始菌株相比分别提高了18. 11%、14. 87%、46. 12%,DHA产量为6. 59 g/L。经5次传代,性状稳定。
A high-DHA-producing strain of the Schizochytrium limacinum SR21 was expected to obtain using the treatment of atmospheric and room temperature plasmas( ARTP),and the screening method of iodoacetic acid,malonic acid and compound plate. After treatment of ARTP for 15 s at 100 W power and10 L/min air flow rate,a mutant strain named NA2 was selected using 1. 2 g/L malonic acid as a screening plate. Cultured in shake flask for 120 h,the biomass,oil productivity and DHA productivity of NA2 strain were 18. 11%,14. 87% and 46. 12% higher than those of original strain,respectively. The DHA productivity reached 6. 59 g/L. The results of subculture test showed that the mutant strain had stable hereditary character after five generations.
A high-DHA-producing strain of the Schizochytrium limacinum SR21 was expected to obtain using the treatment of atmospheric and room temperature plasmas( ARTP),and the screening method of iodoacetic acid,malonic acid and compound plate. After treatment of ARTP for 15 s at 100 W power and10 L/min air flow rate,a mutant strain named NA2 was selected using 1. 2 g/L malonic acid as a screening plate. Cultured in shake flask for 120 h,the biomass,oil productivity and DHA productivity of NA2 strain were 18. 11%,14. 87% and 46. 12% higher than those of original strain,respectively. The DHA productivity reached 6. 59 g/L. The results of subculture test showed that the mutant strain had stable hereditary character after five generations.
引文
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[11]梁园梅,刘瑛,李晶晶,等.高产DHA破囊壶菌Aurantiochytrium sp.PKU#SW7诱变株的筛选[J].微生物学通报,2016(2):457-464.
[12]王婷婷,詹晓北,郑志永,等.高产花生四烯酸高山被孢霉菌株的诱变筛选[J].工业微生物,2013(4):44-50.
[13]王申强.裂殖壶菌产DHA的发酵工艺研究及高产菌株选育[D].江苏无锡:江南大学,2013.
[2]LIANG Y,SARKANY N,CUI Y,et al.Use of sweet sorghum juice for lipid production by Schizochytrium limacinum SR21[J].Bioresour Technol,2010,101(10):3623-3627.
[3]QU L,REN L J,SUN G N,et al.Batch,fed-batch and repeated fed-batch fermentation processes of the marine thraustochytrid Schizochytrium sp.for producing docosahexaenoic acid[J].Bioprocess Biosyst Eng,2013,36(12):1905-1912.
[4]高芳霞,俞岩青,王昆蓉,等.常压室温等离子体及紫外复合诱变选育达托霉素高产菌株[J].中国抗生素杂志,2016(6):425-428.
[5]袁军,赵犇,孙梦玉,等.常压室温等离子体(ARTP)诱变快速选育高产DHA的裂殖壶菌突变株[J].生物技术通报,2015(10):199-204.
[6]LI J,LIU R,CHANG G,et al.A strategy for the highly efficient production of docosahexaenoic acid by Aurantiochytrium limacinum SR21 using glucose and glycerol as the mixed carbon sources[J].Bioresour Technol,2015,177:51-57.
[7]孙佼文,丁健,贾禄强,等.Schizochytrium sp.S31生产DHA底物流加策略及呼吸特性分析[J].中国油脂,2018(2):110-114.
[8]FU J,CHEN T,LU H,et al.Enhancement of docosahexaenoic acid production by low-energy ion implantation coupled with screening method based on Sudan black Bstaining in Schizochytrium sp.[J].Bioresour Technol,2016,221:405-411.
[9]程功,徐建中,郭燕风,等.常压室温等离子体诱变选育L-精氨酸生产菌及发酵条件优化[J].微生物学通报,2016(2):360-369.
[10]李慧玲,刘永梅.诱变选育高产DHA裂殖壶菌突变株[J].食品科技,2015(9):12-16.
[11]梁园梅,刘瑛,李晶晶,等.高产DHA破囊壶菌Aurantiochytrium sp.PKU#SW7诱变株的筛选[J].微生物学通报,2016(2):457-464.
[12]王婷婷,詹晓北,郑志永,等.高产花生四烯酸高山被孢霉菌株的诱变筛选[J].工业微生物,2013(4):44-50.
[13]王申强.裂殖壶菌产DHA的发酵工艺研究及高产菌株选育[D].江苏无锡:江南大学,2013.