亚精胺(spermidine)具有抗衰老、提高记忆力、诱导细胞自噬等多种生物活性,但生物合成量偏低,挖掘亚精胺合成相关基因资源对其代谢工程育种至关重要。在解淀粉芽胞杆菌中,已预测含有亚精胺合成酶基因(speE),然而其在亚精胺形成中的功能尚未得到鉴定。为证实speE基因的功能,研究构建了speE基因缺失菌株和回补表达菌株,探究了该基因对亚精胺合成的影响。结果表明,敲除speE基因未检测到亚精胺,菌株丧失亚精胺合成能力;回补speE基因后,亚精胺合成能力恢复。该研究首次证实了speE基因在解淀粉芽胞杆菌合成亚精胺中的功能,为亚精胺的代谢工程育种及相关研究奠定了基础。
Spermidine is a natural polyamine involved in numerous biological processes, which has the functions of anti-aging, improving memory, and inducing autophagy. However, the low spermidine yield limits its development. It is crucial to excavate the genes related to spermidine synthesis for metabolic engineering breeding. The spermidine synthase gene has been predicted as speE in Bacillus amyloliquefaciens, while its function in spermidine formation has not been identified. To confirm the function of speE, a deletion mutant of speE and its complementary strain were constructed, and the impact of these gene manipulations on spermidine synthesis were analyzed. Our results indicated that the spermidine was not detectable after knocking out of speE, revealing that the deletion mutant strain was deprived of the ability to produce spermidine. After complementary expression of speE, the capacity of spermidine synthesis was recovered. In this study, the speE was first confirmed involved in spermidine synthesis in B. amyloliquefaciens, which provided the foundation for the metabolic engineering breeding of spermidine and related research.
[1] JAGU E,POMEL S,PETHE S,et al.Polyamine-based analogs and conjugates as antikinetoplastid agents[J].European Journal of Medicinal Chemistry,2017,139:982-1 015.
[2] 郑宇宏,范旭红,张云峰,等.大豆新型营养因子亚精胺的研究进展[J].大豆科学,2017,36(4):645-650.
[3] IGARASHI K,KASHIWAGI K.Modulation of cellular function by polyamines[J].Internationl Journal of Biochemistry & Cell Biology,2010,42(1):39-51.
[4] PEGG,E. A.Functions of polyamines in mammals[J].Journal of Biological Chemistry,2016,291(29):14 904-14 912.
[5] MADEO F,BAUER M A,CARMONA-GUTIERREZ D,et al.Spermidine: a physiological autophagy inducer acting as an anti-aging vitamin in humans?[J].Autophagy,2019,15(1):165-168.
[6] EISENBERG T,ABDELLATIF M,SCHROEDER S,et al.Cardioprotection and lifespan extension by the natural polyamine spermidine[J].Nature Medicine,2016,22(12):1 428-1 438.
[7] FRANK M,TOBIAS E,FEDERICO P,et al.Spermidine in health and disease[J].Science,2018,359(6 374):eaan2 788.
[8] EISENBERG T,ABDELLATIF M,ZIMMERMANN A,et al.Dietary spermidine for lowering high blood pressure[J].Autophagy,2017,13(4):767-769.
[9] PIETROCOLA F,CASTOLDI F,KEPP O,et al.Spermidine reduces cancer-related mortality in humans[J].Autophagy,2019,15(2):362-365.
[10] EISENBERG T,KNAUER H,SCHAUER A,et al.Induction of autophagy by spermidine promotes longevity[J].Nature Cell Biology,2009,11(11):1 305-1 314.
[11] GUPTA V K,SCHEUNEMANN L,EISENBERG T,et al.Restoring polyamines protects from age-induced memory impairment in an autophagy-dependent manner[J].Nature Neuroscience,2013,16(10):1 453.
[12] PUCCIARELLI S,MORESCHINI B,MICOZZI D,et al.Spermidine and spermine are enriched in whole blood of nona/centenarians[J].Rejuvenation Research,2012,15(6):590-595.
[13] SCALABRINO G,FERIOLI M E.Polyamines in mammalian ageing: an oncological problem, too? A review[J].Mechanisms of ageing and development,1984,26(2-3):149-164.
[14] STEFAN K,RAIMUND P,PETER W,et al.Higher spermidine intake is linked to lower mortality: a prospective population-based study[J].The American Journal of Clinical Nutrition,2018,108(2):371-380.
[15] SODA K,KANO Y,SAKURAGI M,et al.Long-term oral polyamine intake increases blood polyamine concentrations[J].Journal of Nutritional Science & Vitaminology,2009,55(4):361-366.
[16] ALVAREZ M A,MORENO-ARRIBAS M V.The problem of biogenic amines in fermented foods and the use of potential biogenic amine-degrading microorganisms as a solution[J].Trends in Food Science & Technology,2014,39(2):146-155.
[17] QIAN Z G,XIA X X,LEE S Y.Metabolic engineering of Escherichia coli for the production of putrescine: A four carbon diamine[J].Biotechnology and Bioengineering,2009,104(4):651-662.
[18] SEKOWSKA A,BERTIN P,DANCHIN A.Characterization of polyamine synthesis pathway in Bacillus subtilis 168[J].Molecular Microbiology,1998,29(3):851-858.
[19] KIM S-K,WESTPHELING J.Engineering a spermidine biosynthetic pathway in Clostridium thermocellum results in increased resistance to furans and increased ethanol production[J].Metabolic Engineering,2018,49:267-274.
[20] VALD S-SANTIAGO L,CERVANTES-CH VEZ J A,LE N-RAM REZ C G,et al.Polyamine metabolism in fungi with emphasis on phytopathogenic species[J].Journal of Amino Acids,2012,2012:1-13.
[21] SHAH P,SWIATLO E.A multifaceted role for polyamines in bacterial pathogens[J].Molecular Microbiology,2008,68(1):4-16.
[22] 姚响文,周密,曹颖瑛,等.多胺在微生物中的研究进展[J].中国真菌学杂志,2014,9(2):114-118.
[23] KIM S K,JIN Y S,CHOI I G,et al.Enhanced tolerance of Saccharomyces cerevisiae to multiple lignocellulose-derived inhibitors through modulation of spermidine contents[J].Metabolic Engineering,2015,29:46-55.
[24] KIM S K,JO J H,PARK Y C,et al.Metabolic engineering of Saccharomyces cerevisiae for production of spermidine under optimal culture conditions[J].Enzyme & Microbial Technology,2017,101:30-35.
[25] KERA K,NAGAYAMA T,NANATANI K,et al.Reduction of spermidine content resulting from inactivation of two arginine decarboxylases increases biofilm formation in Synechocystis sp. strain PCC 6803[J].Journal of Bacteriology,2018,200(9):e00 664-e00 617.
[26] FENG J,GU Y,QUAN Y,et al.Improved poly-γ-glutamic acid production in Bacillus amyloliquefaciens by modular pathway engineering[J].Metabolic Engineering,2015,32:106-115.
[27] CHEN X H,KOUMOUTSI A,SCHOLZ R,et al.Comparative analysis of the complete genome sequence of the plant growth-promoting bacterium Bacillus amyloliquefaciens FZB42[J].Nature Biotechnology,2007,25(9):1 007.
[28] JIN P,KANG Z,YUAN P,et al.Production of specific-molecular-weight hyaluronan by metabolically engineered Bacillus subtilis 168[J].Metabolic Engineering,2016,35:21-30.
[29] XU J Z,YAN W L,ZHANG W G.Enhancing menaquinone-7 production in recombinant Bacillus amyloliquefaciens by metabolic pathway engineering[J].Rsc Advances,2017,7(45):28 527-28 534.
