食品与发酵工业2018，Vol.44Issue(2)：8-14,7.DOI:10.13995/j.cnki.11-1802/ts.015693
高分子量腈水合酶工程菌生产烟酰胺的工艺建立
Establishment of the process of nicotinamide produced by Escherichia coil expressing high molecular mass nitrile hydratase
摘要
Abstract
To increase the biomass and the activity of Escherichia coli BL21 (DE3) /pET-24a(+)-nhhBrbsArbsG (BAG) expressing high molecular mass nitrile hydratase from Rhodococcus rhodochrous J1 and establish proper process of whole-cell catalysis of 3-cyanopytidine and high cell-density cultivation,we used orthogonal experiment (L9 (34)) to optimize elements in culture media.After comparison of substrate-flow and substrate fed-batch process,technology of fed-batch was applied to achieve high cell-density cultivation.After the optimization of culture media,the biomass and the specific activity of BAG in the flask reached 4.42 g DCW/L and 30.91 U/mgDCW,respectively.As a more appropriate process,substrate fed-batch process was applied to produce nicotinamide using BAG,and the yield of nicotinamide reached 390 g/L.After the high cell-density cultivation of BAG in 5 L bioreactor,the biomass of cell attained 73.97gDCW/L (OD6o0 =200),the specific activity and the total activity reached 42.70 U/mg DCW and 2813 U/mL,respectively.When high density of BAG after fermentation in the tank was used to catalyze 3-cyanopytidine,537 g/L of nicotinamide was obtained,which was the highest yield of nicotinamide among all the reported production from recombinant E.coil expressing nitrile hydratase.关键词
腈水合酶/大肠杆菌/全细胞催化/底物分批补料/高密度发酵Key words
high molecular mass nitrile hydratase/Escherichia coli/whole-cell catalysis/substrate fed-batch/high cell-density cultivation引用本文复制引用
郭军玲,王哲,刘中美,周哲敏..高分子量腈水合酶工程菌生产烟酰胺的工艺建立[J].食品与发酵工业,2018,44(2):8-14,7.基金项目
国家重点研发计划政府间国际科技创新合作重点专项(2016YFE0127400) （2016YFE0127400）
国家自然科学基金青年科学基金项目(31400078) （31400078）
