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.
GOU Zongqin
,
Yintizhaer·TUERXUNMAIMAITI
,
TIAN Yuan
,
DAI Jianxin
,
AI Lianzhong
,
XIONG Zhiqiang
. Optimization of heterogeneous lycopene synthesis in Escherichia coli using response surface methodology[J]. Food and Fermentation Industries, 2022
, 48(15)
: 110
-117
.
DOI: 10.13995/j.cnki.11-1802/ts.030178
[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.
