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重组大肠杆菌全细胞合成苯基乳酸的研究 被引量:5

Production of phenyllactic acid by using whole-cell recombinant Escherichia coli
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摘要 在E.coli BL21(DE3)中过量表达D-乳酸脱氢酶基因(D-ldh),并优化该重组菌全细胞转化苯丙酮酸钠合成苯基乳酸的条件。通过单因素实验和正交实验优化诱导表达条件,并在此基础上对全细胞转化苯丙酮酸钠合成苯基乳酸进行了优化。结果表明,菌体OD600为1.2时添加IPTG至终浓度0.2mmol/L,25℃诱导4h后收集菌体具有最佳转化活性;最优分批转化条件:p H7.0,8.0g/L苯丙酮酸钠,20g/L葡萄糖,1%(v/v)吐温-80,菌体浓度20g/L(干重),37℃,转速200r/min转化0.5h,苯基乳酸产量和转化率分别达到4.91g/L,56%。在上述优化条件上通过流加苯丙酮酸钠和葡萄糖,经6h转化,苯基乳酸最终产量达到17.23g/L,转化率为54%。研究结果表明该重组大肠杆菌成功转化苯丙酮酸合成苯基乳酸,具有较好的应用前景,为系统化代谢工程改造大肠杆菌生物合成苯基乳酸的进一步研究和应用提供了有用的技术参数。 D- lactate dehydrogenase was expressed in recombinant Escherichia coli BL21( DE3),the expression conditions and bioconversion medium were optimized for the production of PLA with whole- cell transformation.Single factor experiments and orthogonal experiments were used to optimize induction conditions and the transformation conditions with the engineered whole- cells transformation. The optimized induction condition for D- ldh expression was IPTG 0.2mmol / L and inducing for 4h at 25℃ and OD6001.2,the optimized batch reaction conditions in phosphate buffer( pH7.0) were: 8g / L PPA,20 g / L glucose,20 g / L( dry cell weight),1% Tween- 80,37℃,200 r / min for 0.5h with PLA production of 4.91 g / L and conversion ratio of 56%.Based on the above optimized conditions,fed- batch fermentation was conducted by intermittent feed PPA and glucose. After 6h transformation,the final PLA concentration reached 17.23 g / L with the conversion ratio of 54%. Results showed that PPA was efficiently transformed into PLA through engineered E.coli BL21( DE3) / p ET28a- D- ldh.This study not only showed a good industrial application prospect,but also laid a foundation for metabolic engineering of E. coli for the production of PLA.
出处 《食品工业科技》 CAS CSCD 北大核心 2015年第9期147-151,157,共6页 Science and Technology of Food Industry
基金 科技部农业科技成果转化资金项目(2013GB2C100176) 江苏省自然科学青年基金项目(BK20130380) 江苏省高校自然科学研究项目(13KJB550002) 江苏省"六大高峰人才"资助计划项目(NY-021) 苏州市科技支撑计划(SNG201354) 常熟市科技发展计划(CN201220)
关键词 大肠杆菌 D-乳酸脱氢酶 苯基乳酸 全细胞生物转化 Escherichia coil D-lactate dehydrogenase phenyllactic acid whole-cell bioconversion
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参考文献20

  • 1Lavermicocca P, Valerio F, Evidente A, et al.Purification and Characterization of Novel AntifungalCompounds from the Sourdough Lactobacillus plantarum Strain 21B [ J ]. Applied and Environmental Microbiology, 2000,66 ( 9 ) : 4084-4090.
  • 2Tuberoso C I G, Bifulco E, Caboni P, et al. Lumichrome and Phenyllactic Acid as Chemical Markers of Thistle (Galactites tomentosa Moench ) Honey [ J ]. Journal of Agricuhural and Food Chemistry,2011,59( 1 ) :364-369.
  • 3Li XF, Jiang B, Pan BL. Biotransformation of phenylpyruvic acid to phenyllactic acid by growing and resting cells of a Lactobacillus sp[ J].Biotechnology Letters,2007,29:593-597.
  • 4Dieuleveux V, Gueguen M. Antimicrobial effects of D- 3- phenyllactic acid on Listeria monocytogenes in TSB-'fE medium, milk, and cheese [ J ]. Journal of Food Protection, 1998,61 ( 10 ) : 1281-1285.
  • 5Dieuleveux V, Lemarinier S, Gueguen M. Antimicrobial spectrum and target site of D- 3- phenyllactic acid [ J ]. International Journal of Food Microbiology, 1998, 40 ( 3 ) : 177-183.
  • 6Valerio F, Lavermicocca P, Pascale M, et al. Production of phenyllactic acid by lactic acid bacteria: an approach to the selection of strains contributing to food quality and preservation [ J ] .FEMS Microbiology Letters ,2004,233 (2) :289-95.
  • 7Lavermicocca P, Valerio F, Visconti A. Antifungal Activity of Phenyllactic Acid against Molds Isolated from Bakery Products [ J ] . Applied and Environmental Microbiology, 2003, 69 ( 1 ) : 634-640.
  • 8Wang J P,Yoo J S,Lee J H,et al.Effects of phenyllactic acid on production performance, egg quality parameters, and blood characteristics in laying hens [ J ]. The Journal of Applied Poultry Research ,2009,18 ( 2 ) :203-209.
  • 9Wang J P,Lee J H,Yoo J S,et al.Effects of phenyllactic acid on growth performance, intestinal microbiota, relative organ weight, blood characteristics, and meat quality of broiler chicks [ J] .Poultry Science ,2010,89 ( 7 ) : 1549-1555.
  • 10Ebdrup S,Pettersson I, Rasmussen H B, et al.Synthesis and biological and structural characterization of the dual- acting peroxisome proliferator-activated receptor alpha/gamma agonist ragaglitazar[ J ]. Journal of Medicinal Chemistry, 2003,46 ( 8 ) : 1306-17.

二级参考文献26

  • 1Schnurer J.Magnusson J.Antifungal lactic acid bacteria as biopreservatives[J].Trends Food Sci Technol,2005,16:70~78
  • 2Allen K L,Molan P C,Reid G M.A survey of the antibacterial activity of some New Zealand honeys[J].J Pharm Pharmacol,1991,43(12):817~822
  • 3Molan P.Russell K.Non-peroxide antibacterial activity in some New Zealand honeys[J].J Apic Res,1988,27(1):62~67
  • 4Russell K M,Molan P C,Wilkins A L,et al.Identification of some antibaeterial constituents of New Zealand manuka honey[J].J Agric Food Chem,1990,(38):10~13
  • 5Wilkins A L.Lu Y,Molan P C.Extractable organic substances from New Zealand unifloral manuka(Leptosperum scoparium)honeys[J].J Apic Res,1993,32:3~9
  • 6Weston R J,Mitchell K R,Allen K L.Antibacterial phenolic components of New Zealand manuka honey[J].Food Chem,1999,64(3):295~301
  • 7Dieuleveux V,der Van P,Chataud J,et al.Purification and characterization of anti-Listeria compounds produced by Geotrichum candidum[J].Appl Environ Microbiol,1998,64(2):800~803
  • 8Lavermicocca P,Valerio F,Evidente A,et al.Purification and characterization of novel antifungal compounds from the sourdough Lactobacillus plantarum strain 21B[J].Appl Environ Microbiol,2000,66(9):4 084~4 090
  • 9Magnusson J.Antifungal activity of lactic acid bacteria[D].Uppsala,Sweden:Swedish University of Agricultural Sciences,2003
  • 10Dieuleveux V,Lemarinier S,Gueguen M.Antimicrobial spectrum and target site of D-3-phenyllactic acid[J].Intern J Food Microbiol,1998,40(3):177~183

共引文献69

同被引文献72

  • 1李德茂,李从发,刘四新,陈利梅.3-苯基乳酸的研究进展[J].药物生物技术,2004,11(5):344-347. 被引量:14
  • 2鲁绯,孙君社,王丽英,杨森莲.雅致放射毛霉胞外蛋白酶特性的研究[J].中国酿造,2004,23(12):8-10. 被引量:13
  • 3赵锡武,何玉莲.果糖的特性及应用[J].饮料工业,2006,9(4):4-9. 被引量:11
  • 4李兴峰,江波,潘蓓蕾.新型生物防腐剂——苯乳酸在食品中的研究与应用[J].食品与发酵工业,2007,33(5):87-91. 被引量:29
  • 5李建武 萧能庆 余瑞元 等.生物化学实验原理和方法[M].北京:北京大学出版社,1997.390-396.
  • 6Dieuleveux V,Vanderpyl D,Chataud J,et al.Purification and characterization of anti-Listeria compounds produced by Geotrichum candidum[J].Applied and Environmental Microbiology,1998,64(2):800-803.
  • 7Tuberoso C I G,Bifulco E,Caboni P,et al.Lumichrome and phenyllactic acid as chemical markers of thistle(Galactites tomentosa moench)honey[J].Journal of Agricultural and Food Chemistry,2010,59(1):364-369.
  • 8LI X,JIANG B,PAN B.Biotransformation of phenylpyruvic acid to phenyllactic acid by growing and resting cells of a Lactobacillus sp.[J].Biotechnology Letters,2007,29(4):593-597.
  • 9WANG J,YOO J,LEE J,et al.Effects of phenyllactic acid on production performance,egg quality parameters,and blood characteristics in laying hens[J].The Journal of Applied Poultry Research,2009,18(2):203-209.
  • 10WANG J,LEE J,YOO J,et al.Effects of phenyllactic acid on growth performance,intestinal microbiota,relative organ weight,blood characteristics,and meat quality of broiler chicks[J].Poultry Science,2010,89(7):1 549-1 555.

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