Please wait a minute...
 
 
食品与发酵工业  2022, Vol. 48 Issue (15): 110-117    DOI: 10.13995/j.cnki.11-1802/ts.030178
  研究报告 本期目录 | 过刊浏览 | 高级检索 |
响应面法优化大肠杆菌异源合成番茄红素
苟宗芹, 音提扎尔·吐尔逊买买提, 田园, 戴健欣, 艾连中, 熊智强*
(上海理工大学 健康科学与工程学院,上海食品微生物工程研究中心,上海,200093)
Optimization of heterogeneous lycopene synthesis in Escherichia coli using response surface methodology
GOU Zongqin, Yintizhaer·TUERXUNMAIMAITI, TIAN Yuan, DAI Jianxin, AI Lianzhong, XIONG Zhiqiang*
(Shanghai Engineering Research Center of Food Microbiology, School of Health Science and Engineering, University of Shanghai for Science and Technology, Shanghai 200093, China)
下载:  HTML  PDF (3810KB) 
输出:  BibTeX | EndNote (RIS)      
摘要 番茄红素是一种C40类胡萝卜素,具有强抗氧化性和提高免疫力等功效,已广泛应用于食品和化妆品等行业。为提高大肠杆菌异源生产番茄红素效率,筛选出最适大肠杆菌生产菌株Top10,采用单因素试验和响应面法优化发酵培养基,确定番茄红素生产最佳配方为LB培养基中添加果糖4.22 g/L,NH4Cl 0.89 g/L和正十二烷15%(体积分数)。在此条件下,番茄红素得率达到29.3 mg/g DCW,比优化前提高了138%。实时荧光定量PCR分析表明,优化后番茄红素合成关键基因IspGcrtE表达比优化前显著增强(P<0.05),分别提高6.9倍和4.5倍。该研究通过宿主筛选和培养基优化显著提高了番茄红素合成效率,为大肠杆菌高效生产番茄红素奠定基础。
服务
把本文推荐给朋友
加入引用管理器
E-mail Alert
RSS
作者相关文章
苟宗芹
音提扎尔·吐尔逊买买提
田园
戴健欣
艾连中
熊智强
关键词:  大肠杆菌  番茄红素  响应面  Box-Behnken 试验设计    
Abstract: Lycopene, a C40 terpenoid compound, has strong anti-oxidation and immunity enhancement effects, which have been widely used in the food and cosmetics industries. To improve the efficiency of heterologous production of lycopene, this study screened Escherichia coli top10 as the suitable production strain and optimized fermentation medium by single factor experiment and response surface methodology. The optimal medium was medium with 4.22 g/L fructose, 0.89 g/L NH4Cl and 15% n-dodecane. Under this condition, the yield of lycopene reached 29.3 mg/g DCW, which was 138% higher than before optimization. Based on real-time fluorescent quantitative PCR, the expressions of key genes IspG and crtE for lycopene synthesis were significantly enhanced (P<0.05) after optimization, which increased by 6.9 times and 4.5 times, respectively. This study significantly improved the yield of lycopene through host screening and medium optimization, laying the foundation for the efficient production of lycopene in E. coli.
Key words:  Escherichia coli    lycopene    response surface    Box-Behnken design
收稿日期:  2021-11-27      修回日期:  2021-12-22           出版日期:  2022-08-15      发布日期:  2022-09-02      期的出版日期:  2022-08-15
基金资助: 上海市科技兴农项目(2019-02-08-00-07-F01152);国家自然科学基金项目(31972101);上海食品微生物工程研究中心项目(19DZ2281100)
作者简介:  第一作者:硕士研究生(熊智强教授为通信作者,E-mail:xiongzq@hotmail.com)
引用本文:    
苟宗芹,音提扎尔·吐尔逊买买提,田园,等. 响应面法优化大肠杆菌异源合成番茄红素[J]. 食品与发酵工业, 2022, 48(15): 110-117.
GOU Zongqin,Yintizhaer·TUERXUNMAIMAITI,TIAN Yuan,et al. Optimization of heterogeneous lycopene synthesis in Escherichia coli using response surface methodology[J]. Food and Fermentation Industries, 2022, 48(15): 110-117.
链接本文:  
http://sf1970.cnif.cn/CN/10.13995/j.cnki.11-1802/ts.030178  或          http://sf1970.cnif.cn/CN/Y2022/V48/I15/110
[1] LI X, WANG Z X, ZHANG G L, et al.Improving lycopene production in Saccharomyces cerevisiae through optimizing pathway and chassis metabolism[J].Chemical Engineering Science, 2019, 193:364-369.
[2] ALPER H, JIN Y S, MOXLEY J F, et al.Identifying gene targets for the metabolic engineering of lycopene biosynthesis in Escherichia coli[J].Metabolic Engineering, 2005, 7(3):155-164.
[3] XIE W P, LV X M, YE L D, et al.Construction of lycopene-overproducing Saccharomyces cerevisiae by combining directed evolution and metabolic engineering[J].Metabolic Engineering, 2015, 30:69-78.
[4] KIM Y S, LEE J H, KIM N H, et al.Increase of lycopene production by supplementing auxiliary carbon sources in metabolically engineered Escherichia coli[J].Applied Microbiology and Biotechnology, 2011, 90(2):489-497.
[5] JIN W Y, XU X, JIANG L, et al.Putative carotenoid genes expressed under the regulation of Shine-Dalgarno regions in Escherichia coli for efficient lycopene production[J].Biotechnology Letters, 2015, 37(11):2 303-2 310.
[6] 杨帆, 苏卜利, 王永红, 等.启动子对重组大肠杆菌合成番茄红素能力的影响[J].食品与发酵工业, 2020, 46(17):27-32.YANG F, SU B L, WANG Y H, et al.Effects of promoters for lycopene production in engineering Escherichia coli[J].Food and Fermentation Industries, 2020, 46(17):27-32.
[7] NAM H K, CHOI J G, LEE J H, et al.Increase in the production of β-carotene in recombinant Escherichia coli cultured in a chemically defined medium supplemented with amino acids[J].Biotechnology Letters, 2013, 35(2):265-271.
[8] 李文. 果糖提高crtEBI重组大肠杆菌合成番茄红素产量的研究[D].天津:天津科技大学, 2014.LI W.Studies on enhancing lycopene production by fructose in crtEBI recombinant Escherichia coli[D].Tianjin:Tianjin University of Science & Technology, 2014.
[9] DU W, SONG Y J, LIU M Y, et al.Gene expression pattern analysis of a recombinant Escherichia coli strain possessing high growth and lycopene production capability when using fructose as carbon source[J].Biotechnology Letters, 2016, 38(9):1 571-1 577.
[10] 陈艳. 高产番茄红素酿酒酵母的设计构建与发酵过程优化[D].天津:天津大学, 2017.CHEN Y.Design and construction of lycopene overproducing Saccharomyces cerevisiae and fermentation process optimization[D].Tianjin:Tianjin University, 2017.
[11] MUSSAGY C U, GUIMARÃES A A C, ROCHA L V F, et al.Improvement of carotenoids production from Rhodotorula glutinis CCT-2186[J].Biochemical Engineering Journal, 2021, 165:107827.
[12] 李一萌. 基因工程菌发酵与杜氏藻环化酶的阻遏产番茄红素的研究[D].广州:华南理工大学, 2015.LI Y M.Research on lycopene production by optimizing fermentation conditions of genetically engineered bacteria and repressing lycopene cyclase Lyc B1 of Dunalie bardawil[D].Guangzhou:South China University of Technology, 2015.
[13] XIA Y J, CHEN Y, LIU X F, et al.Enhancement of antroquinonol production during batch fermentation using pH control coupled with an oxygen vector[J].Journal of the Science of Food and Agriculture, 2019, 99(1):449-456.
[14] 任东雪, 陈鹏程, 郑璞, 等.外源氧载体和前体L-谷氨酸添加策略提高Bacillus subtilis HB-1发酵产γ-聚谷氨酸[J].食品与生物技术学报, 2020, 39(6):84-92.REN D X, CHEN P C, ZHENG P, et al.Addition of exogenous oxygen carrier and precursor L-glutamic acid enhanced the production of γ-poly glutamic acid by fermentation of Bacillus subtilis HB-1[J].Journal of Food Science and Biotechnology, 2020, 39(6):84-92.
[15] ZUO Z Q, XUE Q, ZHOU J, et al.Engineering Haloferax mediterranei as an efficient platform for high level production of lycopene[J].Frontiers in Microbiology, 2018, 9:2893.
[16] ZHANG S, SONG P, LI S.Application of n-dodecane as an oxygen vector to enhance the activity of fumarase in recombinant Escherichia coli:Role of intracellular microenvironment[J].Brazilian Journal of Microbiology, 2018, 49(3):662-667.
[17] 郭燕, 邓杰, 任志强, 等.响应面优化酿酒酵母与窖泥酯化细菌协同发酵产丁酸乙酯和己酸乙酯[J].食品科学, 2021, 42(10):209-217.GUO Y, DENG J, REN Z Q, et al.Optimization of the production of ethyl hexanoate and ethyl butyrate by cofermentation of Saccharomyces cerevisiae and esterifying bacteria from pit mud of Chinese Baijiu using response surface methodology[J].Food Science, 2021, 42(10):209-217.
[18] 张博, 史永吉, 杨辉, 等.通过发酵优化提高大肠杆菌生产L-半胱氨酸产量[J].食品与发酵工业, 2021, 47(18):175-180.ZHANG B,SHI Y J,YANG H, et al.Enhancement of L-cysteine production in Escherichia coli through fermentation optimization[J].Food and Fermentation Industries, 2021, 47(18):175-180.
[19] WANG J F, XIONG Z Q, LI S Y, et al.Exploiting exogenous MEP pathway genes to improve the downstream isoprenoid pathway effects and enhance isoprenoid production in Escherichia coli[J].Process Biochemistry, 2015, 50(1):24-32.
[20] BANERJEE A, WU Y, BANERJEE R, et al.Feedback inhibition of deoxy-d-xylulose-5-phosphate synthase regulates the methylerythritol 4-phosphate pathway[J].Journal of Biological Chemistry, 2013, 288(23):16 926-16 936.
[1] 樊鹏帅, 杨宗玲, 王俊, 马喜山, 刘义凤, 吴逸民, 周志桥, 曹续东, 夏凯. 桑叶提取物压片糖果粉体造粒工艺优化[J]. 食品与发酵工业, 2022, 48(9): 179-185.
[2] 周海岩, 周斌, 邹树平, 张博, 柳志强. 补料分批发酵条件优化提高D-泛酸的产量[J]. 食品与发酵工业, 2022, 48(7): 8-13.
[3] 赵世光, 谢东宝, 储欣颖, 张宇, 胡子敏, 葛礼涛, 钱森和, 魏明. 混菌发酵制备茶籽多肽及其抗氧化作用[J]. 食品与发酵工业, 2022, 48(6): 147-153.
[4] 李洋, 张稳杰, 韩雨辰, 翟懿雪, 张成林. 生物法合成4-羟基异亮氨酸的代谢工程研究进展[J]. 食品与发酵工业, 2022, 48(5): 281-288.
[5] 高艳蕾, OJANGBA THEODORA, 杨超, 王拙, 张丽, 余群力, 郭兆斌. 超高压技术辅助优化牛皮胶原低脂牛肉饼工艺[J]. 食品与发酵工业, 2022, 48(2): 86-93.
[6] 费莹莹, 张珍, 陈雪琴, 李雯. 肉桂精油抑制肠出血性大肠杆菌O157∶H7活性研究[J]. 食品与发酵工业, 2022, 48(14): 63-67.
[7] 名晓东, 郭自涛, 陈剑雄, 顾正华, 辛瑜, 孙海彦, 关彦明, 张梁. 重组大肠杆菌产卤代烷脱卤酶的发酵条件优化[J]. 食品与发酵工业, 2022, 48(13): 16-24.
[8] 文湘郡, 滕鑫, 丁星宇, 佘竹欣, 李壹, 熊晓辉. 一种新型检测大肠杆菌O157∶H7的免疫磁珠-量子点纳米颗粒的制备和应用[J]. 食品与发酵工业, 2022, 48(13): 48-54.
[9] 周有彩, 肖雪花, 梁志波, 何勇锦, 陈必链. 蛋白质高产株小球藻MBFJNU-17的异养培养基优化[J]. 食品与发酵工业, 2022, 48(13): 108-114.
[10] 刘哲, 叶英, 罗黎霞, 王虹, 张祎睿. 狭果茶藨子营养成分分析与氨基酸提取工艺优化及评价[J]. 食品与发酵工业, 2022, 48(13): 188-195.
[11] 陈怡君, 王晓慧, 陈艳萍, 邓敏, 伊帕尔·开斯尔, 龙元爱, 杨万根. 响应面法优化超声波-微波协同辅助酸法提取猕猴桃皮果胶工艺及果胶理化性质分析[J]. 食品与发酵工业, 2022, 48(13): 238-246.
[12] 王曦, 王楚南, 刘菊英, 张琴. 探究KCl代替NaCl对嗜酸乳杆菌和大肠杆菌的影响[J]. 食品与发酵工业, 2022, 48(11): 36-42.
[13] 孙康, 李军, 张鸣宇, 朱凤妹, 葛超, 阎贺静. 美极梅奇酵母冷冻干燥复合保护剂的优化研究[J]. 食品与发酵工业, 2022, 48(11): 73-79.
[14] 罗光宏, 王海蓉, 崔晶, 张喜峰, 何立杰, 尉蕊. 微波辅助低共熔溶剂提取、部分纯化螺旋藻多糖及其体外生物学活性研究[J]. 食品与发酵工业, 2022, 48(11): 107-113.
[15] 王仁佳, 曹文红, 章超桦, 秦小明, 高加龙, 郑慧娜, 林海生. 华贵栉孔扇贝富硒蛋白粉的酶法制备工艺优化及其营养评价[J]. 食品与发酵工业, 2022, 48(10): 84-92.
[1] LI Xiao-min et a. The influence of feruloyl esterase on filterability of wort[J]. Food and Fermentation Industries, 2017, 43(11): 30 -33 .
[2] WU Xiang-yi et al . Effect of yak milk casein hydrolysates on protein carbonyl content and activity of antioxidant enzymes of oxidative damaged HepG2 cells induced by H2O2[J]. Food and Fermentation Industries, 2017, 43(11): 34 -38 .
[3] ZHANG Dong et al. Effect of different amounts of salt on quality of bacon[J]. Food and Fermentation Industries, 2017, 43(11): 159 .
[4] WANG An-feng et al. Optimization of hydrolysis process of pinctadafucata by response surface method[J]. Food and Fermentation Industries, 2017, 43(11): 165 .
[5] TANG Bin et al. Effects of intermittent microwave processing on fresh-keeping of marinated pork[J]. Food and Fermentation Industries, 2017, 43(11): 191 .
[6] ZOU Yu-feng et al. Review onresearchprogressanddevelopmentofgel-typemeat productsprocessing technology[J]. Food and Fermentation Industries, 2017, 43(11): 232 .
[7] ZHANG Dong et al. Researchprogressonreducing sodium chlorideinmeatproducts[J]. Food and Fermentation Industries, 2017, 43(11): 238 .
[8] . Isolation and identification of anaerobic bacteria in the process of Maotai-flavor liquor brewing[J]. Food and Fermentation Industries, 0, (): 1 .
[9] YU Qing-lin et al. Fermentation optimization of recombinant Yarrowia lipolytica for its efficient succinic acid production[J]. Food and Fermentation Industries, 0, (): 1 .
[10] . Test Study on Extruded Maize without Germ Used as Beer Adjunct[J]. Food and Fermentation Industries, 2002, 28(5): 35 .
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
版权所有 © 《食品与发酵工业》编辑部
地址:北京朝阳区酒仙桥中路24号院6号楼111室
本系统由北京玛格泰克科技发展有限公司设计开发  技术支持:support@magtech.com.cn