产反式肉桂酸内生真菌GRE4的鉴定及发酵条件研究Study on Identification and Fermentation Conditions of Trans-Cinnamic Acid-Producing Endophytic Fungus GRE4
张赫,关昕,祁可香,尤梦瑶,郑春英
摘要(Abstract)：
采用HPLC法，以反式肉桂酸为对照，分析内生真菌GRE4的活性成分；采用琼脂糖扩散法，对内生真菌GRE4进行抑菌活性分析，并采用单因素实验及正交实验对内生真菌GRE4的发酵条件进行优化；采用形态学观察及ITS序列分析法，对内生真菌GRE4进行初步菌种鉴定。结果表明，在内生真菌GRE4发酵液中含有反式肉桂酸；抑菌活性实验结果表明，内生真菌对10种致病菌均有较好的抑制作用，尤其对铜绿假单胞菌的抑菌效果较显著；优化后得到GRE4的最佳发酵工艺条件：培养基碳源为质量浓度2.0%的葡萄糖，氮源为质量浓度1.5%的蛋白胨，初始pH值为7.0,装液量为50%(体积比),种子液接种量为5%(体积比),空气浴振荡器仪器参数为140 r/min、28℃培养9 d。经鉴定，内生真菌GRE4为杂色曲霉(Aspergillus versicolor)。上述研究说明内生真菌GRE4为反式肉桂酸产生菌。
关键词(KeyWords)： 反式肉桂酸;内生真菌;抑菌活性;发酵条件;鉴定
基金项目(Foundation): 黑龙江大学校企合作横向项目(20170223)
作者(Author): 张赫,关昕,祁可香,尤梦瑶,郑春英
参考文献(References)：
- [1]KANG N H,MUKHERJEE S,YUN J W.Trans-cinnamic acid stimulates white fat browning and activates brown adipocytes[J].Nutrients,2019,11(3):577.
- [2]HAZIR S,SHAPIRO-ILAN D I,BOCK C H,et al.Trans-cinnamic acid and Xenorhabdus szentirmaii metabolites synergize the potency of some commercial fungicides[J].Journal of Invertebrate Pathology,2017,145:1-8.
- [3]HSEU Y C,KORIVI M,LIN F Y,et al.Trans-cinnamic acid attenuates UVA-induced photoaging through inhibition of AP-1 activation and induction of Nrf2-mediated antioxidant genes in human skin fibroblasts[J].Journal of Dermatological Science,2018,90(2):123-134.
- [4]SOSTHENE N K,AYESHA J.Multicomponent crystals of p-coumaric acid and trans-ferulic acid:structures and physicochemical properties[J].Journal of Molecular Structure,2021,1244:377-386.
- [5]朱燕莉，王正莉，王卫，等.天然食品防腐剂的抑菌机理研究进展[J].中国调味品，2021,46(9):176-180.
- [6]李艳军，刘飞，于韦，等.反式肉桂酸转化条件的优化[J].食品与发酵工业，2010,36(3):103-107.
- [7]LI Z J,CAI L,MEI R F,et al.A highly efficient transformation of cis- to trans-cinnamic acid derivatives by iodine[J].Tetrahedron Letters,2015,56(52):7197-7200.
- [8]何小英，许东颖，廖正福，等.天然肉桂油深加工研究进展[J].中国调味品，2015,40(10):120-124.
- [9]郭胜男，卢金清，蔡君龙，等.HS-SPME-GC-MS联用分析不同产地肉桂挥发性成分[J].中国调味品，2014,39(12):113-117,128.
- [10]TRICHUR S S.The need to study the holobiome for gainful uses of endophytes[J].Fungal Biology Reviews,2020,34(3):144-150.
- [11]尤梦瑶，万璐，闫佳佳，等.甘草内生菌研究概况[J].中国农学通报，2022,38(26):20-26.
- [12]崔莹莹，李明燕，翟兴伟，等.微生物发酵法制备鲜辣沙蟹汁工艺研究[J].中国调味品，2022,47(3):140-145.
- [13]CALVILLO á,PELLICER T,CARNICER M,et al.Bioprocess strategies for vitamin B12 production by microbial fermentation and its market applications[J].Bioengineering(Basel),2022,9(8):365.
- [14]THUAN N H,TATIPAMULA V B,CANH N X,et al.Recent advances in microbial co-culture for production of value-added compounds[J].Biotechnology,2022,12(5):115.
- [15]LI X J Y,SUN R,LIU R P.Natural products in licorice for the therapy of liver diseases:progress and future opportunities[J].Pharmacological Research,2019,144:210-226.
- [16]施丰成，梁佳欣，刘远上，等.反应时间对以甘草为原料制备香料挥发性成分的影响[J].中国调味品，2022,47(10):79-85.
- [17]曼琼，杨志军，邓毅，等.甘草内生菌的鉴定与药理作用和活性成分研究进展[J].中国临床药理学杂志，2018,34(9):1125-1128.
- [18]陈静，许贞，张雪，等.不同产地甘草内生真菌多样性及分离条件研究[J].药学学报，2019,54(2):373-379.
- [19]杨志军，邓毅，曼琼，等.甘草内生菌代谢物的抗炎作用[J].中国现代应用药学，2018,35(5):633-637.
- [20]BRADER G,COMPANT S,MITTER B,et al.Metabolic potential of endophytic bacteria[J].Current Opinion in Biotechnology,2014,27:30-37.
- [21]高娟娟，贾丽艳，田宇敏，等.枯草芽孢杆菌CGMCC 6624对抗生素耐受性的研究[J].中国调味品，2019,44(3):31-35,41.
- [22]郑雪，徐慧超，李伟，等.东北膜荚黄芪内生真菌的分离及其活性代谢物分析[J].中国新药杂志，2017,26(12):1376-1381.
- [23]许哲祥，刘玉洁，刘松梅，等.基于木脂素生物合成的内生菌发酵五味子研究[J].中国调味品，2020,45(4):34-37.
- [24]张赫，尤梦瑶，万璐，等.产紫丁香苷内生真菌CJ7的鉴定及发酵条件研究[J].中国农学通报，2022,38(25):143-150.
- [25]孙会刚，陈志轩，卞路遥，等.牛蒡和苔干复配提取物的抑菌活性及其防腐保鲜作用[J].中国调味品，2022,47(4):198-202,206.
- [26]万璐，闫佳佳，尤梦瑶，等.产甘草次酸内生真菌GRE111的鉴定及发酵条件研究[J].中国调味品，2022,47(10):41-46.
- [27]STEELE P R,PIRES J C.Biodiversity assessment:state-of-the-art techniques in phylogenomics and species identification[J].American Journal of Botany,2011,98(3):415-425.
- [28]PAREEK C S,SMOCZYNSKI R,TRETYN A.Sequencing technologies and genome sequencing[J].Journal of Applied Genetics,2011,52(4):413-435.
- [29]WANG H N,YANG X H,WEI S H.Analysis of Aspergillus versicolor exudate composition[J].Journal of Basic Microbiology,2022,62(10):1241-1253.
- [30]罗寒，李晓栋，李晓明，等.红树林来源内生真菌杂色曲霉Aspergillus versicolor MA-229次级代谢产物研究[J].中国抗生素杂志，2017,42(4):334-340.
- [31]LI H,ZHANG R,CAO F,et al.Proversilins A-E,drimane-type sesquiterpenoids from the endophytic Aspergillus versicolor[J].Journal of Natural Products,2020,83(7):2200-2206.
- [32]DING Y,ZHU X,HAO L,et al.Bioactive indolyl diketopiperazines from the marine derived endophytic Aspergillus versicolor DY180635[J].Marine Drugs,2020,18(7):338.
- [33]HU J,LI Z,GAO J,et al.New diketopiperazines from a marine-derived fungus strain Aspergillus versicolor MF180151[J].Marine Drugs,2019,17(5):262.
- [34]YANG L J,PENG X Y,ZHANG Y H,et al.Antimicrobial and antioxidant polyketides from a deep-sea-derived fungus Aspergillus versicolor SH0105[J].Marine Drugs,2020,18(12):636.
- [35]YANG F X,DAI J M,LIU H Y,et al.Isochromenes from the Nicotiana tabacum-derived endophytic fungus Aspergillus versicolor and their anti-tobacco mosaic virus activities[J].Natural Product Research,2022,22:1-9.
- [36]许楚旋，王嘉琦，蒋冬花.1株产生洛伐他汀杂色曲霉的筛选、鉴定和发酵优化条件[J].生物技术通报，2018,34(11):210-215.