Abstract: Betaine can effectively improve the fermentation performance of strains in industrial production. The effect of betaine on threonine fermentation and intracellular metabolism of Escherichia coli THRD were investigated. The threonine titer increased from 50.74 g/L to 54.59 g/L, while the yield increased from 37.45% to 40.69% when betaine was fed. In such case, 28 significantly different metabolites related threonine anabolism were discovered. Metabolic flux analysis showed that carbon flow distribution of glycolysis and pentose phosphate pathway were changed, CO2 fixation reaction catalyzed by phosphoenol pyruvate carboxylase and tricarboxylic acid cycle were regulated, and bypass metabolism of glyceraldehyde-3-phosphate, aspartic acid and threonine were reduced, thus providing sufficient reducing power and precursor for threonine synthesis. The results revealed the effect of betaine on E. coli on the metabolic level and provided a theoretical reference for the application of betaine in other industrial fermentation products.
李杰,田俊宇,季圆清,等. 代谢组学分析甜菜碱对大肠杆菌合成苏氨酸的影响[J]. 食品与发酵工业, 2021, 47(17): 34-40.
LI Jie,TIAN Junyu,JI Yuanqing,et al. Effect of betaine on synthesis of threonine in Escherichia coli by metabonomic analysis[J]. Food and Fermentation Industries, 2021, 47(17): 34-40.
[1] BECKER J,WITTMANN C.Systems and synthetic metabolic engineering for amino acid production-the heartbeat of industrial strain development[J].Current Opinion in Biotechnology,2012,23(5):718-726. [2] SONG K H,LEE H H,HYUN H H.Characterization of salt-tolerant mutant for enhancement of L-threonine production in Escherichia coli[J].Applied Microbiology and Biotechnology,2000,54(5):647-651. [3] DONG X Y,QUINN P J,WANG X Y.Metabolic engineering of Escherichia coli and Corynebacterium glutamicum for the production of L-threonine[J].Biotechnology Advances,2011,29(1):11-23. [4] ALMAAS E,KOVÁCS B,VICSEK T,et al.Global organization of metabolic fluxes in the bacterium Escherichia coli[J].Nature,2004,427(6 977):839-843. [5] LEE K H,PARK J H,KIM T Y,et al.Systems metabolic engineering of Escherichia coli for L-threonine production[J].Molecular Systems Biology,2007,3(1):149. [6] LEE J H,SUNG B H,KIM M S,et al.Metabolic engineering of a reduced-genome strain of Escherichia coli for L-threonine production[J].Microbial Cell Factories,2009,8(1):1-12. [7] CHEN N,HUANG J,FENG Z B,et al.Optimization of fermentation conditions for the biosynthesis of L-threonine by Escherichia coli[J].Applied Biochemistry and Biotechnology,2009,158(3):595-604. [8] SU Y W,GUO Q Q,WANG S,et al.Effects of betaine supplementation on L-threonine fed-batch fermentation by Escherichia coli[J].Bioprocess and Biosystems Engineering,2018,41(10):1 509-1 518. [9] FIGUEROA-SOTO C G,VALENZUELA-SOTO E M.Glycine betaine rather than acting only as an osmolyte also plays a role as regulator in cellular metabolism[J].Biochimie,2018,147:89-97. [10] XIA W,PENG W F,CHEN W,et al.Interactive performances of betaine on the metabolic processes of Pseudomonas denitrificans[J].Journal of Industrial Microbiology and Biotechnology,2015,42(2):273-278. [11] WARREN M J,RAUX E,SCHUBERT H L,et al.The biosynthesis of adenosylcobalamin (vitamin B12)[J].Natural Product Reports,2002,19(4):390-412. [12] YING H X,HE X,LI Y,et al.Optimization of culture conditions for enhanced lysine production using engineered Escherichia coli[J].Applied Biochemistry and Biotechnology,2014,172(8):3 835-3 843. [13] XU K,XU P.Betaine and beet molasses enhance L-lactic acid production by Bacillus coagulans[J].PLoS One,2014,9(6).DOI:10.1371/journal.pone.0100731. [14] 刘旭峰, 王宁,郝亚男,等.CRISPRi干扰中心代谢基因转录对苏氨酸合成的影响[J].食品与发酵工业,2019,45(8):1-7. LIU X F,WANG N,HAO Y N,et al.Threonine synthesis under interfered transcriptions of genes involved in central metabolic pathway by CRISPRi[J].Food and Fermentation Industries,2019,45(8):1-7. [15] MI Z H,KWOK L Y,XUE J G,et al.Fermentation dynamics of Lactobacillus helveticus H9 revealed by ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry[J].International Journal of Food Science & Technology,2018,53(6):1 442-1 451. [16] 苏跃稳. L-苏氨酸基因工程菌的改造及发酵过程的优化[D].长春:吉林大学,2017. SU Y W.Modification of L-threonine-producing strain and optimization of fermentation process[D].Changchun:Jilin University,2017. [17] LI Y J,ZHANG D Z,CAI N Y,et al.Betaine supplementation improved l-threonine fermentation of Escherichia coli THRD by upregulating zwf (glucose-6-phosphate dehydrogenase) expression[J].Electronic Journal of Biotechnology,2019,39:67-73. [18] 郭群群. 三甲基甘氨酸对大肠杆菌发酵生产L-苏氨酸的影响[D].长春:吉林大学,2018. GUO Q Q.Effects of trimethylglycine on L-threonine production in Escherichia coli[D].Changchun:Jilin University,2018.