内生真菌DBR-9发酵液中抗菌物质分析及发酵条件
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摘要
[目的]研究内生真菌DBR-9发酵液中橘霉素的HPLC检测方法及橘霉素的发酵条件。[方法]通过文献调研及条件试验确定橘霉素HPLC分析条件,通过单因素和正交试验确定最佳发酵条件。[结果]橘霉素HPLC分析条件为C18柱,流动相为甲醇和磷酸水溶液,样品液为发酵液-甲醇(体积比1∶9)。DBR-9最佳发酵条件为初始接种量10.4×106个孢子,26℃、160 r/min振摇培养30 d。[结论]运用HPLC方法可精确测定DBR-9发酵液中橘霉素含量,发酵条件的确定对利用DBR-9生产橘霉素有重要参考价值。
[Aims] The aim was to study the HPLC detection method of antimicrobial substance citrinin in fermentation broth of endophytic fungus DBR-9 and its fermentation conditions. [Methods] The conditions of HPLC analysis were determined by literature research and condition experiments, and the fermentation conditions of endophytic fungus DBR-9 were determined by single factor and orthogonal test. [Results] The HPLC analysis conditions of citrinin were C18 column, the mobile phase was methanol and phosphoric acid water solution, and the sample solution was fermentation broth and methanol with volume ratio of 1∶9. The optimal fermentation conditions of DBR-9 were initial inoculation of 10.4×106 spores, and cultured at 26 ℃ and 160 r/min for 30 d. [Conclusions] The HPLC method can accurately determine the content of citrinin in endophytic fungus DBR-9 fermentation broth. The confirmation of optimal fermentation conditions have important reference value for large-scale production of citrinin using endophytic fungus DBR-9.
[Aims] The aim was to study the HPLC detection method of antimicrobial substance citrinin in fermentation broth of endophytic fungus DBR-9 and its fermentation conditions. [Methods] The conditions of HPLC analysis were determined by literature research and condition experiments, and the fermentation conditions of endophytic fungus DBR-9 were determined by single factor and orthogonal test. [Results] The HPLC analysis conditions of citrinin were C18 column, the mobile phase was methanol and phosphoric acid water solution, and the sample solution was fermentation broth and methanol with volume ratio of 1∶9. The optimal fermentation conditions of DBR-9 were initial inoculation of 10.4×106 spores, and cultured at 26 ℃ and 160 r/min for 30 d. [Conclusions] The HPLC method can accurately determine the content of citrinin in endophytic fungus DBR-9 fermentation broth. The confirmation of optimal fermentation conditions have important reference value for large-scale production of citrinin using endophytic fungus DBR-9.
引文
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[12]许艳丽,吴振兴,何晓霞,等.免疫亲和柱净化-高效液相色谱法测定红曲制品中桔霉素的含量[J].食品安全质量检测学报,2016,7(11):4649-4652.
[13]陈蕴,许赣荣,顾玉梅,等.TLC及HPLC测定红曲产品中的桔霉素[J].食品与生物技术学报,2001,20(2):164-168.
[2]梁彩云,王森胜,何熙璞,等.阴香内生真菌Y-74鉴定及其次级代谢产物的初步研究[J].基因组学与应用生物学,2015,34(4):775-780.
[3]高彤,蔡思源,许兰兰,等.海洋来源真菌Penicillim grisefulvum中桔霉素类化合物的分离及抗菌活性[J].高等学校化学学报,2016,37(7):1282-1286.
[4]ODHAV B,NAICKER V.Mycotoxins in South African Traditionally Brewed Beers[J].Food Additives&Contaminants,2002,19(1):55-61.
[5]SHU Pin-yen,LIN Cheng-huang.Simple and Sensitive Determination of Citrinin in Monascus by GC-Selected Ion Monitoring Mass Spectrometry[J].Analytical Sciences,2002,18(3):283.
[6]CHEN Fu-sheng,HU Xiao-qing.Study on Red Fermented Rice with High Concentration of Monacolin K and Low Concentration of Citrinin[J].International Journal of Food Microbiology,2005,103(3):331-337.
[7]李楠,陈俊霖,顾双,等.霉菌发酵食品中桔霉素的免疫亲和柱结合高效液相荧光法检测[J].中国调味品,2018,43(4):33-37.
[8]陈旭峰,周立平,孙佰申,等.红曲霉所产桔霉素及其检测与控制[J].酿酒,2004,31(4):113-116.
[9]潘振球,冯家力,王一红,等.液相色谱-串联质谱法检测食品中的桔霉素[J].中国卫生检验杂志,2008,18(11):2192-2193.
[10]陈福生,邢淑婕.红曲产品中桔霉素含量的ELISA测定[J].食品科学,2004,25(8):169-172.
[11]张晓伟,王加华,李培睿.红曲米中桔霉素、淀粉、水分含量的快速测定[J].中国粮油学报,2017,32(6):152-158.
[12]许艳丽,吴振兴,何晓霞,等.免疫亲和柱净化-高效液相色谱法测定红曲制品中桔霉素的含量[J].食品安全质量检测学报,2016,7(11):4649-4652.
[13]陈蕴,许赣荣,顾玉梅,等.TLC及HPLC测定红曲产品中的桔霉素[J].食品与生物技术学报,2001,20(2):164-168.
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