灵菌红素具有抗氧化、抗肿瘤等生物活性和潜在的医用价值,大规模生产对其研究和开发具有重要意义。从槐树根际土壤中筛选出1株产红色素菌株H04,根据16S rDNA系统发育树鉴定该菌株为黏质沙雷氏菌Serratia marcesence。其合成的红色素经UV、HPLC-MS分析,确定为灵菌红素。利用单因素试验和正交试验优化发酵条件,以提高色素产量。最佳发酵条件为:培养基使用15 g/L蔗糖为碳源、0.5 g/L酵母浸粉和0.5 g/L(NH4)2SO4为氮源、5 g/L NaCl、0.75 g/L KCl、10 g/L脯氨酸、0.5 g/L CaCO3;接种量3%(体积分数)、温度25 ℃、转速180 r/min、培养时间24 h。优化后发酵得灵菌红素提取物4.5 g/L,与正交优化前相比产量提高了2.3倍。
The large-scale production of prodigiosin, which has biological activities such as antioxidant and antitumor, is of great importance for its research and development. A red pigment producing bacteria H04, isolated from the rhizosphere of Sophora japonica Linn., was identified as Serratia marcescens according to the 16S rDNA phylogenetic tree. The red pigment derived from H04 was determined as prodigiosin through UV and HPLC-MS. Single-factor and orthogonal experiments were used to optimize the fermentation conditions to improve the pigment yield. The optimum conditions were the fermentation medium using 15 g/L sucrose as carbon source, 0.5 g/L yeast extract powder, and 0.5 g/L ammonium sulfate as nitrogen source, 5 g/L NaCl, 0.75 g/L KCl, 10 g/L proline, and 0.5 g/L CaCO3; inoculum amount was 3%, 25 ℃, 180 r/min, 24 h. The optimized fermentation yielded 4.5 g/L of red pigment extract, with a 2.3-fold increase in yield compared to that before orthogonal optimization.
[1] HAN R, XIANG R J, LI J L, et al.High-level production of microbial prodigiosin:A review[J].Journal of Basic Microbiology, 2021, 61(6):506-523.
[2] KAVITHA R, AISWARIYA S, RATNAVALI C M G.Anticancer activity of red pigment from Serratia marcescens in human cervix carcinoma[J].International Journal of PharmTech Research, 2010, 2(1):784-787.
[3] CHIU W J, LIN S R, CHEN Y H, et al.Prodigiosin-emerged PI3K/beclin-1-independent pathway elicits autophagic cell death in doxorubicin-sensitive and-resistant lung cancer[J].Journal of Clinical Medicine, 2018, 7(10):321.
[4] YIP C H, MAHALINGAM S, WAN K L, et al.Prodigiosin inhibits bacterial growth and virulence factors as a potential physiological response to interspecies competition[J].PLoS One, 2021, 16(6):e0253445.
[5] SHAIKH Z.Biosynthesis of prodigiosin and its applications[J].IOSR Journal of Pharmacy and Biological Sciences, 2016, 11:1-28.
[6] DARSHAN N, MANONMANI H K.Prodigiosin inhibits motility and activates bacterial cell death revealing molecular biomarkers of programmed cell death[J].AMB Express, 2016, 6(1):1-12.
[7] LIU W J, YANG J, TIAN Y N, et al.An in situ extractive fermentation strategy for enhancing prodigiosin production from Serratia marcescens BWL1001 and its application to inhibiting the growth of Microcystis aeruginosa[J].Biochemical Engineering Journal, 2021, 166:107836.
[8] HAN S B, PARK S H, JEON Y J, et al.Prodigiosin blocks T cell activation by inhibiting interleukin-2R alpha expression and delays progression of autoimmune diabetes and collagen-induced arthritis[J].The Journal of Pharmacology and Experimental Therapeutics, 2001, 299(2):415-425.
[9] DARSHAN N, MANONMANI H K.Prodigiosin and its potential applications[J].Journal of Food Science and Technology, 2015, 52(9):5 393-5 407.
[10] KIMYON Ö, DAS T, IBUGO A I, et al.Serratia secondary metabolite prodigiosin inhibits Pseudomonas aeruginosa biofilm development by producing reactive oxygen species that damage biological molecules[J].Frontiers in Microbiology, 2016, 7:972.
[11] LAPENDA J L, ALVES V P, ADAM M L, et al.Cytotoxic effect of prodigiosin, natural red pigment, isolated from Serratia marcescens UFPEDA 398[J].Indian Journal of Microbiology, 2020, 60(2):182-195.
[12] HERRÁEZ R, MUR A, MERLOS A, et al.Using prodigiosin against some gram-positive and gram-negative bacteria and Trypanosoma cruzi[J].The Journal of Venomous Animals and Toxins Including Tropical Diseases, 2019, 25:e20190001.
[13] FARAAG A, EL-BATAL A, EL-HENDAWY H.Characterization of prodigiosin produced by Serratia marcescens strain isolated from irrigation water in Egypt[J].Nature and Science, 2017, 15(5):55-68.
[14] WEI Y H, YU W J, CHEN W C.Enhanced undecylprodigiosin production from Serratia marcescens SS-1 by medium formulation and amino-acid supplementation[J].Journal of Bioscience and Bioengineering, 2005, 100(4):466-471.
[15] WEI Y H, CHEN W C.Enhanced production of prodigiosin-like pigment from Serratia marcescens SMΔR by medium improvement and oil-supplementation strategies[J].Journal of Bioscience and Bioengineering, 2005, 99(6):616-622.
[16] 张丹峰, 杨培周, 操丽丽, 等.粘质沙雷氏菌摇瓶发酵产灵菌红素的工艺优化[J].食品科学, 2015, 36(13):119-124.
ZHANG D F, YANG P Z, CAO L L, et al.Optimization of shake flask cultivation conditions for prodigiosin production by Serratia marcescens[J].Food Science, 2015, 36(13):119-124.
[17] BHAGWAT A, PADALIA U.Optimization of prodigiosin biosynthesis by Serratia marcescens using unconventional bioresources[J].Journal, Genetic Engineering & Biotechnology, 2020, 18(1):26.
[18] 陈新颜, 操丽丽, 杨培周, 等.响应面试验优化黏质沙雷氏菌利用菜籽饼粕产灵菌红素工艺[J].食品科学, 2017, 38(4):231-236.
CHEN X Y, CAO L L, YANG P Z, et al.Response surface optimization of prodigiosin production of Serratia marcescens using rapeseed meal[J].Food Science, 2017, 38(4):231-236.
[19] WILLIAMSON N R, FINERAN P C, LEEPER F J, et al.The biosynthesis and regulation of bacterial prodiginines[J].Nature Reviews Microbiology, 2006, 4(12):887-899.
[20] LEWIS S M, CORPE W A.Prodigiosin-producing bacteria from marine sources[J].Applied Microbiology, 1964, 12(1):13-17.
[21] LI D, LIU J, WANG X, et al.Biological potential and mechanism of prodigiosin from Serratia marcescens subsp.lawsoniana in human choriocarcinoma and prostate cancer cell lines[J].International Journal of Molecular Sciences, 2018, 19(11):3465.
