添加不同表面活性剂对胞苷发酵的影响
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摘要
研究4种不同表面活性剂对大肠杆菌BG-12的胞苷合成和菌体生长的影响。在胞苷产生菌大肠杆菌BG-12发酵10 h时,添加Tween-80、Triton X-100、CTAB (hexadecyl trimethyl ammonium bromide)和EMB(ethambutol dihydrochloride) 4种表面活性剂,进行对比试验,分析菌体生长、耗糖速率和胞苷浓度等的变化。Tween-80的添加量为1～4 g/L时,对胞苷的产量呈现促进作用,添加3 g/L时的产量达到最大为1.696 g/L,是对照组的2.42倍,且对菌体生长、耗糖速率无明显影响;添加Triton X-100时实验组的胞苷产量均低于对照组,但添加量为0.4～0.8 g/L时会促进菌体生长和耗糖速率;添加CTAB和EMB时,胞苷产量明显下降,但对菌体生长及耗糖速率影响不大。Tween-80在发酵10 h时添加、且在低于4 g/L时对胞苷产量有促进作用,添加Triton X-100、CTAB和EMB对菌体生长表现为促进或无影响,但对胞苷产量无促进作用。
The effects of four different surfactants,Tween-80,Triton X-100,CTAB,and EMB,on cytidine synthesis by fermentation with Escherichia coli BG-12 as well as cell grouth were studied. The yield of cytidine increased while 1-4 g/L Tween-80 was added. The maximum yield of 1.7 g/L was obtained when 3 g/L of Tween-80 was added,which was 2.42 times higher than that of the control. Additionally,it had no significant effects on bacteria growth and sugar consumption. When TritonX-100 was added,the yield of cytidine decreased,however,0.4-0.8 g/L TritonX-100 could promote bacteria growth and sugar consumption. When CTAB or EMB was added,the yield of cytidine decreased obviously,but had little effects on bacteria growth and sugar consumption. In conclusion,the yield of cytidine increased when less than 4 g/L Tween-80 was added after 10 h fermentation. In comparison,TritonX-100,CTAB,and EMB inhibited cytidine production,although they might stimulate cell growth.
The effects of four different surfactants,Tween-80,Triton X-100,CTAB,and EMB,on cytidine synthesis by fermentation with Escherichia coli BG-12 as well as cell grouth were studied. The yield of cytidine increased while 1-4 g/L Tween-80 was added. The maximum yield of 1.7 g/L was obtained when 3 g/L of Tween-80 was added,which was 2.42 times higher than that of the control. Additionally,it had no significant effects on bacteria growth and sugar consumption. When TritonX-100 was added,the yield of cytidine decreased,however,0.4-0.8 g/L TritonX-100 could promote bacteria growth and sugar consumption. When CTAB or EMB was added,the yield of cytidine decreased obviously,but had little effects on bacteria growth and sugar consumption. In conclusion,the yield of cytidine increased when less than 4 g/L Tween-80 was added after 10 h fermentation. In comparison,TritonX-100,CTAB,and EMB inhibited cytidine production,although they might stimulate cell growth.
引文
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[22]段莹莹,赵伟,曹大鹏,等.表面活性剂对葡萄糖酸钠发酵的影响[J].食品工业,2014(10):118-120.
[2]方海田,谢希贤,徐庆阳,等.微生物发酵法生产胞嘧啶核苷的研究进展[J].发酵科技通讯,2010,39(3):48-51.
[3]朱晖.枯草芽孢杆菌胞苷生产菌株的构建[D].天津:天津大学,2013.
[4]吴晓娇,孙家凯,霍文婷,等.过表达car AB和pyr BI对大肠杆菌发酵胞苷的影响[J].中国生物工程杂志,2012,32(2):39-44.
[5]ANDERSON E,SALISBURY V.Rapid in-vitro testing for chemotherapy sensitivity in leukaemia patients[M]//Bioluminescence:Fundamentals and Applications in Biotechnology-Volume 2.Springer Berlin Heidelberg,2014:189-214.
[6]魏志强,徐庆阳,刘淑云.枯草芽孢杆菌生产胞苷的途径分析[J].现代食品科技,2008,24(6):544-547.
[7]FANG H T,XIE X X,XU Q Y,et al.Enhancement of cytidine production by coexpression of gnd,zwf,and prs genes in recombinant Escherichia coli CYT15[J].Biotechnology Letters,2013,35(2):245-251.
[8]FANG H T,LIU H Y,CHEN N,et al.Site-directed mutagenesis studies on the uridine monophosphate binding sites of feedback inhibition in carbamoyl phosphate synthetase and effects on cytidine production by Bacillus amyloliquefaciens[J].Canadian Journal of Microbiology,2013,59(6):374-379.
[9]王梦娇,方海田,刘慧燕,等.转运蛋白Nup C与Nup G微生物胞内核苷分泌的作用研究进展[J].生物技术通报,2017,33(2):24-29.
[10]吕哲喆,刘晓侠,孙诗清,等.表面活性剂对天然红色素发酵的影响[J].药物生物技术,2012(4):324-327.
[11]赵世光,李德才,薛正莲,等.表面活性剂对活跃链霉菌产那西肽发酵的影响[J].中国抗生素杂志,2015,40(5):338-343.
[12]LIANG Y,ZHU L,GAO M,et al.Influence of Tween-80 on the production and structure of water-insoluble curdlan from Agrobacterium sp.[J].International Journal of Biological Macromolecules,2018,106:611-619.
[13]SHENG L,ZHU G,TONG Q.Mechanism study of Tween 80enhancing the pullulan production by Aureobasidium pullulans[J].Carbohydrate Polymers,2013,97(1):121-123.
[14]GALINDO E,SALCEDO G.Detergents improve xanthan yield and poly-mer quality in cultures of Xanthomonas campestris[J].Enzyme and Microbial Technology,1996,19(2):145-149.
[15]YANG H,XIAO C,MA W,et al.The production of hypocrellin colorants by submerged cultivation of the medicinal fungus Shiraia bambusicola[J].Dyes and Pigments,2009,82(2):142-146.
[16]CAI Y,LIAO X,LIANG X,et al.Induction of hypocrellin production by Triton X-100 under submerged fermentation with Shiraia sp.SUPER-H168[J].New Biotechnology,2011,28(6):588-592.
[17]温琦,刘登如,陈坚,等.添加表面活性剂促进兽疫链球菌高产透明质酸[J].化工进展,2006,25(9):1 089-1 094.
[18]YANG Z,XU X,YANG C H,et al.Total synthesis of nannocystin A[J].Organic Letters,2016,18(21):5 768-5 770.
[19]LIU J,CHEN X,DAI X,et al.Effect of surfactant on the production of thermophilic protease from Bacillus stearothermophilus[J].Journal of Microbiology,2004,24(6):58-61.
[20]XIA Z.Effect of Tween 80 on the production of curdlan by Alcaligenes faecalis ATCC 31749[J].Carbohydrate Polymers,2013,98(1):178-180.
[21]卫云路,宁正祥,郑成.新型表面活性剂在谷氨酸发酵中的应用[J].现代食品科技,2009,25(3):289-295.
[22]段莹莹,赵伟,曹大鹏,等.表面活性剂对葡萄糖酸钠发酵的影响[J].食品工业,2014(10):118-120.