Influence of temperature on the production of monascin and ankaflavin by Monascus ruber M7 during liquid state fermentation

doi:10.3969/j.issn.1000-1565.2023.04.009

Abstract

Abstract: To improve the production of monascin(MS)and ankaflavin(AF)by Monascus ruber M7, the changes in composition and yield of alcohol-soluble pigments produced at different fermentation temperatures were investigated using liquid state fermentation. It was found that pigments produced at lower temperatures(25、30 ℃)were orange in hue, with a maximum absorption around 470 nm, and- DOI:10.3969/j.issn.1000-1565.2023.04.009温度对红色红曲菌M7液态发酵产monascin和ankaflavin的影响徐勇¹,邱子娅¹,陈莎^1,2,张佳兰³,高梦祥^1,2,李利^1,2(1.长江大学生命科学学院,湖北荆州 434025;2.长江大学食品研究院,湖北荆州 434025;3.长江大学动物科学学院,湖北荆州 434025)摘要:为了提高红色红曲菌(Monascus ruber)M7发酵生产红曲素(monascin, MS)和红曲黄素(ankaflavin, AF)的产量,采用液态发酵法,探究不同发酵温度下其产生的醇溶性色素在组成和产量方面的变化规律.结果发现,当温度较低时(25、30 ℃),所产生的色素在470 nm有最大吸收,色调偏橙,红斑素(rubropunctatin, RP)和红曲红素(monascorubrin, MR)为优势组分;当温度较高时(35、40 ℃),所产生的色素在400 nm 附近有最大吸收,色调偏黄,MS和AF成为优势组分.随着温度的升高,所有色素组分的产量均先升高后降低,其中,25、40 ℃不利于色素的产生,产量显著低于30、35 ℃;30 ℃时,RP和MR的产量达到最大值,分别为107、124 mg/L,而MS和AF的产量较低,分别为48、26 mg/L;35 ℃时,MS和AF的产量达到最大值,分别为100、47 mg/L,而RP和MR的产量相对30 ℃时显著降低,分别为57、50 mg/L.qPCR分析发现,温度对MS和AF液态发酵的影响与其对色素合成关键酶基因的转录调控密切相关,其中,对色素合成分支节点关键酶基因MrPigF和MrPigH的差异调节,可能对35 ℃时MS和AF产量的显著提高起到了较关键的作用.该结果可为提高MS和AF液态发酵产量提供参考.关键词:红色红曲菌;红曲素;红曲黄素;液态发酵;温度中图分类号:TS201.3 文献标志码:A 文章编号:1000-1565(2023)04-0408-12Influence of temperature on the production of monascin and ankaflavin by Monascus ruber M7 during liquid state fermentationXU Yong¹,QIU Ziya¹,CHEN Suo^1,2,ZHANG Jialan³,GAO Mengxiang^1,2,LI Li^1,2(1. College of Life Science, Yangtze University, Jingzhou 434025, China; 2. Institute of Food Science and Technology, Yangtze University, Jingzhou 434025, China; 3. College of Animal Science, Yangtze University, Jingzhou 434025, China)Abstract: To improve the production of monascin(MS)and ankaflavin(AF)by Monascus ruber M7, the changes in composition and yield of alcohol-soluble pigments produced at different fermentation temperatures were investigated using liquid state fermentation. It was found that pigments produced at lower temperatures(25、30 ℃)were orange in hue, with a maximum absorption around 470 nm, and- 收稿日期:2022-06-07 基金项目:国家自然科学基金资助项目(31730068) 第一作者:徐勇(1997—),男,四川达州人,长江大学在读硕士研究生,主要从事红曲色素的发酵及其在食品中的应用研究. E-mail:1903437308@qq.com 通信作者:李利(1983—),女,湖北浠水人,长江大学副教授,博士,主要从事真菌次生代谢产物及其在食品中的应用研究. E-mail:lily2012@yangtzeu.edu.cn第4期徐勇等:温度对红色红曲菌M7液态发酵产monascin和ankaflavin的影响rubropunctatin(RP)and monascorubrin(MR)were the dominant components; while pigments produced at higher temperatures(35、40 ℃)were yellow in hue, with a maximum absorption around 400 nm, and MS and AF became the dominant ones. As the temperature increased, the yields of each pigment compounds first increased and then decreased. The yields of all pigment compounds were significantly lower at 25、40 ℃ than at 30、35 ℃, indicating that these temperatures were not conducive to the production of pigments. At 30 ℃, RP and MR had a maximum yield of 107、124 mg/L, respectively, while MS and AF had a lower yield, being 48、26 mg/L, respectively. At 35 ℃, MS and AF reached a maximum yield of 100、47 mg/L, while RP and MR decreased significantly when compared to 30 ℃, being 57、50 mg/L, respectively. qPCR analysis revealed that the influence of temperature on the liquid state fermentation of MS and AF was closely related to its transcriptional regulation on key genes responding to pigments biosynthesis. The differential regulation on MrPigF and MrPigH, the key enzyme genes of the pigments biosynthesis branch node, might playe a critical role in improving the yields of MS and AF at 35 ℃. These results would be helpful for the promotion of MS and AF during liquid state fermentation.

Key words: Monascus ruber, monascin, ankaflavin, liquid state fermentation, temperature

CLC Number:

TS201.3

XU Yong,QIU Ziya,CHEN Suo,ZHANG Jialan,GAO Mengxiang,LI Li. Influence of temperature on the production of monascin and ankaflavin by Monascus ruber M7 during liquid state fermentation[J]. Journal of Hebei University(Natural Science Edition), 2023, 43(4): 408-419.

References

[1] CHEN W P, FENG Y L, MOLNáR I, et al. Nature and nurture: confluence of pathway determinism with metabolic and chemical serendipity diversifies Monascus azaphilone pigments[J]. Nat Prod Rep, 2019, 36(4): 561-572. DOI: 10.1039/c8np00060c.
[2] CHEN W P, CHEN R F, LIU Q P, et al. Orange, red, yellow: biosynthesis of azaphilone pigments in Monascus fungi[J]. Chem Sci, 2017, 8(7): 4917-4925. DOI: 10.1039/C7SC00475C.
[3] 张薄博,管宏伟,陈磊,等.红曲黄色素的多样性及产品标准的讨论[J].食品与发酵工业, 2018, 44( 12): 261-266. DOI: 10.13995/j.cnki.11-1802/ts.017008.
[4] 刘彩,陈莎,高梦祥,等.红曲橙色素的发酵制备及其在红曲红色素化学半合成中的应用[J].食品研究与开发, 2021, 42(6): 57-62. DOI: 10.12161/j.issn.1005-6521.2021.06.011.
[5] ADIN S N, GUPTA I, PANDA B P, et al. Monascin and ankaflavin—biosynthesis from Monascus purpureus, production methods, pharmacological properties: a review[J]. Biotech and App Biochem, 2023, 70: 137-147. DOI: 10.1002/bab. 2336.
[6] CHEN G, WU Z Q. Production and biological activities of yellow pigments from Monascus fungi[J]. World J Microbiol Biotechnol, 2016, 32(8): 136-143. DOI: 10.1007/s11274-016-2082-8.
[7] 陈莎,刘涛,王帅,等.红曲黄色素和红曲橙色素的稳定性研究[J].食品工业科技, 2021, 42(3): 19-24. DOI: 10.13386/j.issn1002-0306.2020040208.
[8] LIN C H, LIN T H, PAN T M. Alleviation of metabolic syndrome by monascin and ankaflavin: the perspective of Monascus functional foods[J]. Food Funct, 2017, 8(6): 2102-2109. DOI: 10.1039/c7fo00406k.
[9] LIU S F, WANG Y R, SHEN Y C, et al. A randomized, double-blind clinical study of the effects of Ankascin 568 plus on blood lipid regulation[J]. J Food Drug Anal, 2018, 26(1): 393-400. DOI: 10.1016/j.jfda.2017.04.006.
[10] LEE C L, LIN P Y, HSU Y W, et al. Monascus-fermented monascin and ankaflavin improve the memory and learning ability in amyloid β-protein intracerebroventricular-infused rat via the suppression of Alzheimer’s disease risk factors[J]. J Funct Foods, 2015, 18: 387-399. DOI: 10.1016/j.jff.2015.08.002.
[11] YONGSMITH B, THONGPRADIS P, KLINSUPA W, et al. Fermentation and quality of yellow pigments from golden brown rice solid culture by a selected Monascus mutant[J]. Appl Microbiol Biotechnol, 2013, 97(20): 8895-8902. DOI: 10.1007/s00253-013-5106-4.
[12] 况嘉铀,屈廷啟,韦胜,等.不同温度对红曲霉液态发酵合成红曲黄色素及桔霉素的影响[J].中国调味品, 2021, 46(11): 8-11. DOI: 10.3969/j.issn.1000-9973.2021.11.002.
[13] 毛鹏.红曲黄色素液态发酵技术研究[D].无锡: 江南大学, 2015.
[14] 管宏伟,刘婷婷,陈磊,等.红曲菌液态发酵产天然黄色素的条件优化[J].食品与发酵工业, 2020, 46(2): 150-156. DOI: 10.13995/j.cnki.11-1802/ts.022088.
[15] 赵会.液态发酵生产红曲黄色素过程优化研究[D].长沙: 中南林业科技大学, 2021.
[16] 周波,王菊芳,吴振强,等.高产红曲黄色素菌株的选育[J].微生物学通报, 2008, 35(12): 1909-1914.
[17] FENG Y L, SHAO Y C, CHEN F S. Monascus pigments[J]. Appl Microbiol Biotechnol, 2012, 96(6): 1421-1440. DOI: 10.1007/s00253-012-4504-3.
[18] HUANG T, TAN H L, CHEN G, et al. Rising temperature stimulates the biosynthesis of water-soluble fluorescent yellow pigments and gene expression in Monascus ruber CGMCC10910[J]. AMB Express, 2017, 7(1): 134-143. DOI: 10.1186/s13568-017-0441-y.
[19] LI L, CHEN S, GAO M X, et al. Acidic conditions induce the accumulation of orange Monascus pigments during liquid-state fermentation of Monascus ruber M7[J]. Appl Microbiol Biotechnol, 2019, 103(20): 8393-8402. DOI: 10.1007/s00253-019-10114-8.
[20] QIU Z, ZHANG J, CHEN S, et al. Preparation of extracellular and intracellular water-insoluble Monascus pigments during submerged fermentaion[J]. Appl Biochem Microbiol, 2020, 56(6): 692-698. DOI: 10.1134/S0003683820060149.
[21] CHEN G, BEI Q, HUANG T, et al. Variations in Monascus pigment characteristics and biosynthetic gene expression using resting cell culture systems combined with extractive fermentation[J]. Appl Microbiol Biotechnol, 2018, 102(1): 117-126. DOI: 10.1007/s00253-017-8576-y.
[22] 范菲,段雅丽,刘亚鹏,等.鉴定红曲菌中萘醌响应基因提高红曲色素产率[J].食品与发酵工业, 2022, 48(5): 35-40, 46. DOI: 10.13995/j.cnki.11-1802/ts.028183.
[23] CHEN S, SU D X, GAO M X, et al. A facile macroporous resin-based method for separation of yellow and orange Monascus pigments[J]. Food Sci Biotechnol, 2021, 30(4): 545-553. DOI: 10.1007/s10068-021-00892-1.
[24] 杨洋.红曲色素的发酵、提取及其稳定性研究[D].兰州: 西北师范大学, 2018.
[25] 邱鹏.红曲霉高密度培养及红曲色素发酵工艺研究[D].成都: 西华大学, 2015.
[26] 代斌.红曲色素液态发酵工艺研究[D].成都: 西华大学, 2013.
[27] PATROVSKY M, SINOVSKA K, BRANSKA B, et al. Effect of initial pH, different nitrogen sources, and cultivation time on the production of yellow or orange Monascus purpureus pigments and the mycotoxin citrinin[J]. Food Sci Nutr, 2019, 7(11): 3494-3500. DOI: 10.1002/fsn3.1197.
[28] HUANG Z B, ZHANG S Y, XU Y, et al. Development of an HPLC-UV detection method for the simultaneous determination of two Monascus orange pigments in red yeast rice[J]. Food Anal Methods, 2016, 9(1): 148-155. DOI: 10.1007/s12161-015-0185-8.
[29] LIU L J, ZHANG X H, WANG Z L. Highly efficient production of tailored Monascus pigments by using a biocompatible chemical reaction interfacing with microbial metabolism[J]. ACS Sustainable Chem Eng, 2021, 9(8): 3347-3356. DOI: 10.1021/acssuschemeng.0c09422.
[30] XIONG X, ZHANG X H, WU Z Q, et al. Accumulation of yellow Monascus pigments by extractive fermentation in nonionic surfactant micelle aqueous solution[J]. Appl Microbiol Biotechnol, 2015, 99(3): 1173-1180. DOI: 10.1007/s00253-014-6227-0.
[31] DUAN Y L, JIA L L, PEI X L, et al. An efficient microbial-based method for production of high-purity Monascus azaphilones pigments[J]. LWT, 2022, 170: 114053. DOI: 10.1016/j.lwt.2022.114053.
[32] BALAKRISHNAN B, PARK S H, KWON H J. Inactivation of the oxidase gene mppG results in the selective loss of orange azaphilone pigments in Monascus purpureus[J]. Appl Biol Chem, 2017, 60(4): 437-446. DOI: 10.1007/s13765-017-0296-6. (