溶氧对L-羟脯氨酸发酵的影响及其控制
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摘要
为了研究溶氧对L-羟脯氨酸发酵的影响,分别在不同的溶氧水平下进行发酵实验,通过对菌体生长情况、L-羟脯氨酸产量、糖酸转化率和副产物乙酸积累量的分析,确定L-羟脯氨酸发酵的最佳溶氧控制条件.结果表明:采用分阶段溶氧控制策略,在发酵前期(0~8h)维持溶氧在20%~30%,发酵中期(8~24h)维持溶氧在30%~40%,发酵后期(24~32h)溶氧维持在20%~30%.在此条件下进行L-羟脯氨酸发酵时,L-羟脯氨酸产量和糖酸转化率最高,分别达到45.3g/L和20.7%,并且乙酸积累量较低,仅为1.7g/L.结果表明,分阶段溶氧控制策略既有利于菌体生长,促进产物高效合成,同时又能有效降低副产物乙酸的形成,使整个L-羟脯氨酸发酵过程维持良好的状态.
In order to study on the effect of dissolved oxygen on L-hydroxyproline fermentation,different concentrations of dissolved oxygen was investigated.Through analysis of cell growth,L-hydroxyproline production,glucose conversion rate and by-product acetate accumulation,the optimal dissolved oxygen control conditions for L-hydroxyproline fermentation were determined.The results showed that concentration of L-hydroxyproline reached 45.3 g/L,glucose conversion was 20.7%,and the accumulation of acetate was keep at a lower level of 1.7 g/L,under the condition of staged dissolved oxygen control strategy(in the early stage of fermentation(0~8 h),dissolved oxygen was maintained at 20% ~30%;in the medium stage of fermentation(8~24 h),dissolved oxygen was controlled at 30%~40%;in the late stage of fermentation(24~32 h),dissolved oxygen was maintained at 20% ~30%).It could be concluded that dissolved oxygen stage control strategy was not only beneficial to growth of bacteria and biosynthesis of the product,but could also effectively reduce the formation of acetic acid,keeping the entire Lhydroxyproline fermentation process at a good state.
In order to study on the effect of dissolved oxygen on L-hydroxyproline fermentation,different concentrations of dissolved oxygen was investigated.Through analysis of cell growth,L-hydroxyproline production,glucose conversion rate and by-product acetate accumulation,the optimal dissolved oxygen control conditions for L-hydroxyproline fermentation were determined.The results showed that concentration of L-hydroxyproline reached 45.3 g/L,glucose conversion was 20.7%,and the accumulation of acetate was keep at a lower level of 1.7 g/L,under the condition of staged dissolved oxygen control strategy(in the early stage of fermentation(0~8 h),dissolved oxygen was maintained at 20% ~30%;in the medium stage of fermentation(8~24 h),dissolved oxygen was controlled at 30%~40%;in the late stage of fermentation(24~32 h),dissolved oxygen was maintained at 20% ~30%).It could be concluded that dissolved oxygen stage control strategy was not only beneficial to growth of bacteria and biosynthesis of the product,but could also effectively reduce the formation of acetic acid,keeping the entire Lhydroxyproline fermentation process at a good state.
引文
[1]刘合栋.高产反式-4-羟脯氨酸重组大肠杆菌的构建和发酵优化[D].无锡:江南大学,2013.
[2] SHIBASAKI T,HASHIMOTO S,MORI H,et al.Construction of a novel hydroxyproline-producing recombinant Escherichia coli,by introducing aproline 4-hydroxylase gene[J].Journal of bioscience&bioengineering,2000,90(5):522-525.
[3] SHIBASAKI T,MORI H,OZAKI A.Enzymatic production of trans-4-hydroxy-L-proline by regio-and stereospecific hydroxylation of L-proline[J].Bioscience biotechnology&biochemistry,2000,64(4):746-750.
[4]胡丹丹.无外源L-脯氨酸产反式-4-羟脯氨酸大肠杆菌的构建及发酵优化[D].无锡:江南大学,2015.
[5]张自强,赵东旭,杨新林.羟脯氨酸的研究与开发[J].氨基酸和生物资源,2006,28(1):55-58.
[6] JURCZAK J,PROKOPOWICZ P,GOLEBIOWSKI A.Highly stereoselective synthesis of cis-(2R,3S)-3-hydroxyproline[J].Tetrahedron letters,1993,34(44):7107-7110.
[7]陈宁,范晓光.我国氨基酸产业现状及发展对策[J].发酵科技通讯,2017,46(4):193-197.
[8]陈宁,范晓光.氨基酸生产菌株的研究热点及发展动向[J].发酵科技通讯,2016,45(1):1-6.
[9] ZHAO T X,LI M,ZHENG X,et al.Improved production of trans-4-hydroxy-L-proline by chromosomal integration of the Vitreoscilla hemoglobin gene into recombinant Escherichia coli with expression of proline-4-hydroxylase[J].Journal of bioscience&bioengineering,2016,123(1):109.
[10]杨梦晨,刘镇瑜,陈宁,等.精氨酸对Escherichia coli发酵生产L-色氨酸的影响[J].发酵科技通讯,2017,46(1):16-19.
[11]杨梦晨,户红通,蔡萌萌,等.L-色氨酸清液发酵工艺研究[J].食品与发酵科技,2017,53(2):29-33..
[12]蔡萌萌,户红通,刘子强,等.活细胞在线监控L-羟脯氨酸补料发酵工艺的研究[J].食品与发酵工业,2018,44(5):10-15.
[13]张兆峰,孙丽美,张现峰.溶氧对氨基酸发酵的影响分析[J].科技与创新,2016(10):98-98.
[14] MAJEWSKI R A,DOMACH M M.Simple constrained-optimization view of acetate overflow in E.coli[J].Biotechnology&bioengineering,1990,35(7):732.
[15]徐庆阳,冯志彬,孙玉华,等.溶氧对L-苏氨酸发酵的影响[J].微生物学通报,2007,34(2):312-314.
[16]刘晓波,李宗伟,闫世梁,等.溶氧控制对氨基酸发酵的影响[J].安徽农业科学,2008(19):7977-7979.
[2] SHIBASAKI T,HASHIMOTO S,MORI H,et al.Construction of a novel hydroxyproline-producing recombinant Escherichia coli,by introducing aproline 4-hydroxylase gene[J].Journal of bioscience&bioengineering,2000,90(5):522-525.
[3] SHIBASAKI T,MORI H,OZAKI A.Enzymatic production of trans-4-hydroxy-L-proline by regio-and stereospecific hydroxylation of L-proline[J].Bioscience biotechnology&biochemistry,2000,64(4):746-750.
[4]胡丹丹.无外源L-脯氨酸产反式-4-羟脯氨酸大肠杆菌的构建及发酵优化[D].无锡:江南大学,2015.
[5]张自强,赵东旭,杨新林.羟脯氨酸的研究与开发[J].氨基酸和生物资源,2006,28(1):55-58.
[6] JURCZAK J,PROKOPOWICZ P,GOLEBIOWSKI A.Highly stereoselective synthesis of cis-(2R,3S)-3-hydroxyproline[J].Tetrahedron letters,1993,34(44):7107-7110.
[7]陈宁,范晓光.我国氨基酸产业现状及发展对策[J].发酵科技通讯,2017,46(4):193-197.
[8]陈宁,范晓光.氨基酸生产菌株的研究热点及发展动向[J].发酵科技通讯,2016,45(1):1-6.
[9] ZHAO T X,LI M,ZHENG X,et al.Improved production of trans-4-hydroxy-L-proline by chromosomal integration of the Vitreoscilla hemoglobin gene into recombinant Escherichia coli with expression of proline-4-hydroxylase[J].Journal of bioscience&bioengineering,2016,123(1):109.
[10]杨梦晨,刘镇瑜,陈宁,等.精氨酸对Escherichia coli发酵生产L-色氨酸的影响[J].发酵科技通讯,2017,46(1):16-19.
[11]杨梦晨,户红通,蔡萌萌,等.L-色氨酸清液发酵工艺研究[J].食品与发酵科技,2017,53(2):29-33..
[12]蔡萌萌,户红通,刘子强,等.活细胞在线监控L-羟脯氨酸补料发酵工艺的研究[J].食品与发酵工业,2018,44(5):10-15.
[13]张兆峰,孙丽美,张现峰.溶氧对氨基酸发酵的影响分析[J].科技与创新,2016(10):98-98.
[14] MAJEWSKI R A,DOMACH M M.Simple constrained-optimization view of acetate overflow in E.coli[J].Biotechnology&bioengineering,1990,35(7):732.
[15]徐庆阳,冯志彬,孙玉华,等.溶氧对L-苏氨酸发酵的影响[J].微生物学通报,2007,34(2):312-314.
[16]刘晓波,李宗伟,闫世梁,等.溶氧控制对氨基酸发酵的影响[J].安徽农业科学,2008(19):7977-7979.