小叶榕抗菌活性内生菌发酵条件的优化
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摘要
为了优化小叶榕抗菌活性内生菌的发酵条件,提高其发酵液生物量和抗菌物质的量。笔者采用单因子试验对小叶榕内生细菌QYPT-B01菌株发酵条件优化,确定了QYPT-B01菌株在本试验中的最优发酵条件为:玉米粉3%,豆饼粉4%,K2HPO4·3H2O 0.3%,初始p H 8.0,装液量30%,接种量1%,摇床转速160 r/min,培养温度34℃。在上述条件下培养48 h,发酵液中生物量达到峰值,测得OD值为7.95,与优化前相比较,OD值提高了52.0%(优化前OD值为5.23)。培养72 h,抗菌物质产量达到峰值,此时发酵滤液对铜绿假单胞菌的抑菌圈直径为26.42 mm,与优化前相比较,抑菌圈直径提高了13.5%(优化前抑菌圈直径为23.28 mm)。在优化的发酵条件下,QYPT-B01菌株发酵液的生物量和抗菌物质的量均有明显增多,为后续分离纯化、鉴定其发酵滤液中的抗菌物质,探究其抑菌机制,开发生防菌剂或新型抗生素奠定基础。
To optimize the fermentation conditions of endophytes with antibacterial activity separated from Ficus microphylla, and increase the biomass of fermentation broth and the amount of antibacterial substances,the optimal fermentation condition of the QYPT-B01 strain was optimized by single factor test. The results showed that: the optimum fermentation condition of the QYPT-B01 strain in this experiment was as follows:3% corn flour, 4% bean cake powder, 0.3% K_2HPO_4 · 3H_2O, initial pH value 8.0, loading amount 30%,fermentation inoculation amount 1%, shaking speed 160 r/min, culture temperature 34℃. Under the condition,the biomass reached the maximum value after culturing 48 h, the OD value of fermentation liquid was 7.95,which increased by 52.0% compared with 5.23 before optimization. The amount of antibacterial substances reached the maximum value after culturing 72 h, the bacteriostasis diameter of fermentation filtrate to Pseudomonas aeruginosa was 26.42 mm, which increased by 13.5% compared with 23.28 mm before optimization. Under the optimized fermentation condition, the biomass of the QYPT-B01 strain and the amount of antibacterial substances increase significantly, which lay a foundation for further separation and purification,identification of antibacterial substances in fermentation filtrate, exploration of the antibacterial mechanism,and the development of antimicrobial agents or new antibiotics.
To optimize the fermentation conditions of endophytes with antibacterial activity separated from Ficus microphylla, and increase the biomass of fermentation broth and the amount of antibacterial substances,the optimal fermentation condition of the QYPT-B01 strain was optimized by single factor test. The results showed that: the optimum fermentation condition of the QYPT-B01 strain in this experiment was as follows:3% corn flour, 4% bean cake powder, 0.3% K_2HPO_4 · 3H_2O, initial pH value 8.0, loading amount 30%,fermentation inoculation amount 1%, shaking speed 160 r/min, culture temperature 34℃. Under the condition,the biomass reached the maximum value after culturing 48 h, the OD value of fermentation liquid was 7.95,which increased by 52.0% compared with 5.23 before optimization. The amount of antibacterial substances reached the maximum value after culturing 72 h, the bacteriostasis diameter of fermentation filtrate to Pseudomonas aeruginosa was 26.42 mm, which increased by 13.5% compared with 23.28 mm before optimization. Under the optimized fermentation condition, the biomass of the QYPT-B01 strain and the amount of antibacterial substances increase significantly, which lay a foundation for further separation and purification,identification of antibacterial substances in fermentation filtrate, exploration of the antibacterial mechanism,and the development of antimicrobial agents or new antibiotics.
引文
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[20]张红娟,卢海波,赵立娟,等.内生菌HT5产生抗菌物质培养基及发酵条件优化[J].山西农业科学,2014,54(5):57-61.
[21]胡陈云,李勇,刘敏,等.枯草芽孢杆菌ge25菌株发酵及其抑菌活性成分的初步研究[J].中国中药杂志,2014,39(14):2624-2626.
[2]邓雪萍,杨博.药用植物内生菌及其产生的活性成分研究现状[J].安徽农学通报,2010,16(1):78-80.
[3]陈雪英,都晓伟,李滨.内生菌与药用植物活性成分相关性研究进展[J].国外医药.植物药分册,2008,23(2):47-51.
[4]张玲琪,王海昆,邵华,等.美登木内生真菌产抗癌物质球毛壳甲素的分离及鉴定[J].中国药学杂志,2002,37(3):172-175.
[5]陈至里,沈娟,王影姣,等.美洲大蠊肠道内生细菌的分离及其初步抑菌活性研究[J].生物学通报,2016,51(11):54-57.
[6]邓振山,魏婷婷,苏瑞,等.一株具有抑菌活性的酸枣内生菌的分离[J].浙江农业学报,2017,29(12):2068-2076.
[7]艾嫦,朱妍妍,赵长琦.乌头属植物化学成分、药理作用及其内生菌的研究进展[J].天然产物研究与开发,2012,24(2):248-259.
[8]黄晓琴,张丽霞,刘会香,等.抗茶树冰核细菌内生菌的筛选及鉴定[J].茶叶科学,2015,52(1):97-102.
[9]隆丽林,易有金,周金伟,等.虎杖内生菌转化虎杖苷和转化产物抗氧化活性研究[J].现代食品科技,2015,31(5):76-83.
[10]贾力维.植物内生真菌代谢产物研究进展[J].黑龙江科技信息,2016,20(35):96.
[11]李晓璐.苦参杀虫活性菌株的筛选[J].广东化工,2016,43(12):83-84.
[12]牛丽丽,袁肖寒,顾成波,等.内生真菌产植物次生代谢产物研究进展[J].安徽农业科学,2016,56(11):20-24.
[13]王志伟,纪燕玲,陈永敢.植物内生菌研究及其科学意义[J].微生物学通报,2015,42(2):126-140.
[14]徐萌,王金缘,胡金丽,等.植物内生菌对大豆促生长和抗胁迫作用的研究进展[J].农业科技,2017,36(6):965-969.
[15]邓雪萍.抗菌活性植物内生菌的分离、筛选及发酵条件优化[D].广州:华南理工大学,2010:30-38.
[16]林爱玉,邢晓科,郭顺星,等.4种药用半红树植物内生真菌的分离及其抗菌活性研究[J].中国药学杂志,2006,41(12):892-894.
[17]周晓坤,陈钧,韩邦兴,等.水菖蒲内生菌分离与抗菌活性菌株筛选[J].食品科学,2010,31(9):211-215.
[18]李文鹏,蔡磊,毕廷菊,等.胨冻样芽孢杆菌(Bacillus mucilaginosus)的分离、筛选及其发酵培养基研究[J].土壤通报,2001,45(2):70-72.
[19]陈建华,刘佳佳,臧巩固,等.紫杉醇产生菌的筛选与发酵条件的调控[J].中南大学学报:自然科学版,2004,35(1):65-69.
[20]张红娟,卢海波,赵立娟,等.内生菌HT5产生抗菌物质培养基及发酵条件优化[J].山西农业科学,2014,54(5):57-61.
[21]胡陈云,李勇,刘敏,等.枯草芽孢杆菌ge25菌株发酵及其抑菌活性成分的初步研究[J].中国中药杂志,2014,39(14):2624-2626.