微生物发酵青蒿叶和叶渣的研究

doi:10.13560/j.cnki.biotech.bull.1985.2020-0493

摘要/Abstract

摘要：

为扩大青蒿原料的应用途径,延伸青蒿产业链,对青蒿叶和叶渣进行发酵研究。拟开发可用于动物保健的青蒿来源的产品。采用微生物发酵青蒿及青蒿叶渣,检测枯草芽孢杆菌、酿酒酵母菌、植物乳杆菌等菌株发酵青蒿叶和叶渣后其粗蛋白、粗脂肪、粗纤维素以及青蒿素、青蒿乙素、双氢青蒿酸、青蒿酸含量变化。青蒿叶发酵产物及功效成分含量与对照组比较,酵母菌发酵后粗蛋白提高43.05%,植物乳杆菌发酵后粗脂肪提高106%,枯草芽孢杆菌发酵后粗纤维降低43.30%,黑曲霉菌发酵后青蒿素和青蒿乙素分别提高133.27%和88.06%,地衣芽孢杆菌发酵后青蒿酸提高21.49%,枯草芽孢杆菌发酵后双氢青蒿酸提高86.01%;青蒿叶渣发酵产物与对照组比较,植物乳杆菌发酵后粗脂肪提高87.73%,酿酒酵母菌发酵后粗蛋白提高85.30%,枯草芽孢杆菌发酵后粗纤维降低55.67%,枯草芽孢杆菌发酵后双氢青蒿酸提高71.91%,地衣芽孢杆菌发酵后青蒿乙素提高94.71%。微生物发酵青蒿叶和叶渣显著提高其功效成分含量,增加了探索青蒿叶和叶渣发酵产物作为动物保健品的可能性。

关键词: 青蒿原料, 菌株发酵, 含量变化

Abstract:

To expand the application of Artemisia annua raw materials and extend the A. annua industrial chain,the fermentation of A. annua leaves and leaf residues was studied,aiming to develop products derived from A. annua for animal health. In this study we conducted microbial fermentation of A. annua and A. annua leaf residue,and detected content changes of crude protein,crude fat,crude cellulose,artemisinin,artemisinin B,dihydroartemisinic acid,and artemisinic acid after fermentation of A. annua leaves and leaf residue by Bacillus subtilis,yeast,Lactobacillus plantarum and other strains. The contents of the functional components in the fermented product of A. annua leaves were compared with those in the control group,crude protein increased by 43.05% after yeast fermentation,crude fat increased by 106% after L. plantarum fermentation,crude fiber decreased by 43.30% after L. casei fermentation,artemisinin and artemisinin B increased by 133.27% and 88.06% respectively after Aspergillus niger fermentation,artemisinic acid increased by 21.49% after Bacillus licheniformis fermentation,and dihydroartemisinic acid increased by 86.01% after Bacillus subtilis fermentation. Compared A. annua leaf residue fermented products with the control group,crude fat increased by 87.73% after B. subtilis fermentation,crude protein increased by 85.30% after L. plantarum fermentation,crude fiber decreased by 55.67% after T. reesei fermentation,and dihydroartemisinic acid increased by 71.91% after B. subtilis fermentation,artemisinin increased by 94.71% after B. licheniformis fermentation. Microbial fermentation of A. annua leaves and leaf residues can significantly increase the contents of its effective ingredients,and increase the feasibility of exploring the fermentation products of A. annua leaves and leaf residues as animal health products.

Key words: Artemisia annua raw material, strain fermentation, content change

丁嘉伟, 苏新堯, 尹薪项, 马婷玉, 王晚晴, 向丽, 冯宝民, 王彩霞. 微生物发酵青蒿叶和叶渣的研究[J]. 生物技术通报, 2021, 37(2): 63-70.

DING Jia-wei, SU Xin-yao, YIN Xin-xiang, MA Ting-yu, WANG Wan-qing, XIANG Li, FENG Bao-min, WANG Cai-xia. Study on Microbial Fermentation of Artemisia annua Leaves and Leaf Residues[J]. Biotechnology Bulletin, 2021, 37(2): 63-70.

图/表 6

参考文献 24

[1]	丁丹丹. 黄花蒿中四种青蒿素类化合物含量的时空动态研究[D]. 北京:中国中医科学院, 2019. pmid: 29561814
	Ding DD. Study on the spatiotemporal dynamics of four artemisinin compounds in artemisia annua[J]. Beijing:China Academy of Chinese Medical Sciences, 2019. doi: 10.3390/medsci6020026 URL pmid: 29561814
[2]	蒲昭和. 青蒿的功效及应用[N]. 中国中医药报, 2016-06-13.
	Pu ZH. The efficacy and application of Artemisia annua[N]. China News of Traditional Chinese Medicine, 2016-06-13.
[3]	吴泽全. 青蒿—饲料与良药[J]. 四川农业科技, 1990(4):27.
	Wu ZQ. Artemisia annua-feed and good medicine[J]. Science and Technology of Sichuan Agriculture, 1990(4):27.
[4]	Chaturvedi D, Goswami A, Saikia PP, et al. Artemisinin and its derivatives:a novel class of anti-malarial and anti-cancer agents[J]. Chemical Society Reviews, 2010,39:435-454. URL pmid: 20111769
[5]	马琰, 马庆涛, 承南, 等. 青蒿素与抗菌增效剂联合抗鸡球虫效果评价[J]. 中国家禽, 2016,38(2):53-55.
	Ma Y, Ma QT, Cheng N, et al. Evaluation of anti-chicken coccidiosis effect of artemisinin and antibacterial synergist[J]. China Poultry, 2016,38(2):53-55.
[6]	唐雯, 严明. 里氏木霉分泌族的预测及分析[J]. 微生物学报, 2008,48(4):473-479.
	Tang W, Yan M. Prediction and analysis of Trichoderma reesei secretion family[J]. Acta Microbiologica Sinica, 2008,48(4):473-479.
[7]	袁靖宇, 蔡俊杰, 张千里, 等. 毕赤酵母发酵研究进展[J]. 医学信息, 2011,24(2):340-343.
	Yuan JY, Cai JJ, Zhang QL, et al. Research progress of Pichia pastoris fermentation[J]. Medical Information, 2011,24(2):340-343.
[8]	肖仔君, 钟瑞敏, 陈惠音, 等. 植物乳杆菌的生理功能与应用[J]. 中国食品添加剂, 2005,37(3):22-27.
	Xiao ZJ, Zhong MR, Chen HY, et al. Physiological function and application of Lactobacillus plantarum[J]. China Food Additives, 2005,37(3):22-27.
[9]	张菊, 李金敏, 张志焱, 等. 地衣芽孢杆菌的研究进展[J]. 中国饲料, 2012,17(3):9-11.
	Zhang J, Li JM, Zhang ZY, et al. Research progress of Bacillus licheniformis[J]. China Feed, 2012,17(3):9-11.
[10]	张超凤, 汪雨晨, 陈卫平, 等. 柚子皮黑曲霉的分离鉴定及产酶特性研究[J]. 微生物学杂志, 2017,37(3):22-27.
	Zhang CF, Wang YC, Chen WP, et al. Isolation and identification of Aspergillus niger and its enzyme production characteristics[J]. Journal of Microbiology, 2017,37(3):22-27.
[11]	王学东, 李彪, 戴晋军. 活性干酵母在养猪中的应用[J]. 营养与饲料, 2009,16(4):9-10.
	Wang XD, Li B, Dai JJ. Application of active dry yeast in pig breeding[J]. Nutrition and Feed, 2009,16(4):9-10.
[12]	杨锋, 章亭洲. 枯草芽孢杆菌生物学特性的研究[J]. 科技动态, 2011,15(3):34.
	Yang F, Zhang TZ. Study on the biological characteristics of Bacillus subtilis[J]. Science and Technology News, 2011,15(3):34.
[13]	曹瑞博, 汪建明. 干酪乳杆菌的功能性研究及其应用[J]. 中国食品添加剂, 2010,18(6):169-172.
	Cao RB, Wang JM. Functional study and application of Lactobacillus casei[J]. China Food Additives, 2010,18(6):169-172.
[14]	黄萍, 钟新民, 李必元, 等. 雪菜中粗纤维含量的测定[J]. 东北农业科学, 2011,36(1):45-47.
	Huang P, Zhong XM, Li BY, et al. Determination of crude fiber content in Chinese cabbage[J]. Journal of Northeast Agricultural Science, 2011,36(1):45-47.
[15]	汪红, 魏红, 许超. 饲料中粗脂肪含量测定方法的比较研究[J]. 中国畜牧兽医, 2004,31(7):17-18.
	Wang H, Wei H, Xu C. Comparative study on determination methods of crude fat content in feed[J]. Chinese Animal Husbandry and Veterinary Medicine, 2004,31(7):17-18.
[16]	刘燕, 张平, 倪燕, 等. 双缩脲法测定大豆乳清废水中蛋白质含量[J]. 大豆通报, 2006,85(6):24-26.
	Liu Y, Zhang P, Ni Y, et al. Determination of protein content in soybean whey wastewater by biuret method[J]. Soybean Bulletin, 2006,85(6):24-26.
[17]	余正文, 王伯初, 杨占南, 等. 青蒿中青蒿酸、青蒿素及去氧青蒿素同时提取及检测[J]. 药物分析杂志, 2011,31(6):1020-1023.
	Yu ZW, Wang BC, Yang ZN, et al. Simultaneous extraction and detection of artemisinin, artemisinin and deoxyartemisinin in Artemisia annua[J]. Chinese Journal of Pharmaceutical Analysis, 2011,31(6):1020-1023.
[18]	Tian N, Li J, Liu S, et al. Simultaneous isolation of artemisinin and its precursors from Artemisia annua L. by preparative RP-HPLC[J]. Biomed Chromatogr 2012,26:708-713.
[19]	赵帅, 段明房, 胡红伟, 等. 杜仲叶的功能及在养殖动物中的应用[J]. 饲料博览, 2017(3):27.
	Zhao S, Duan MF, Hu HW, et al. The function of Eucommia ulmoides leaves and its application in breeding animals[J]. Feed Review, 2017(3):27.
[20]	乔宏兴, 史洪涛, 刘畅, 等. 发酵中药在养猪业上的研究进展[J]. 中国猪业, 2018,13(3):48-51, 55.
	Qiao HX, Shi HT, Liu C, et al. Research progress of fermented traditional Chinese medicine in pig industry[J]. China Swine Industry, 2018,13(3):48-51, 55.
[21]	何玉华, 赵凤明. 固态发酵水飞蓟残渣对雉鸡生长性能的影响[J]. 中国兽医杂志, 2014,50(11):56-58.
	He YH, Zhao FM. Effect of solid fermented milk thistle residue on pheasant growth performance[J]. Chinese Journal of Veterinary Medicine, 2014,50(11):56-58.
[22]	杜瑞芝. 饲养奶牛常用的饲料分类及其营养价值[J]. 山东畜牧兽医, 2020,41(2):12-13.
	Du RZ. Feed classification and nutritional value of dairy cows[J]. Shangdong Journal of Animal Science and Veterinary Medicine, 2020,41(2):12-13.
[23]	邓侃, 李祖旭, 姜玫秀, 等. 益生微生物在健胃消食片药渣开发中的应用[J]. 江西农业大学学报, 2015,37(4):738-742.
	Deng K, Li ZX, Jiang MX, et al. Application of probiotic microorganisms in the development of medicine residues for Jianweixiaoshi tablets[J]. Acta Agriculturae Universitatis Jiangxiensis, 2015,37(4):738-742.
[24]	李漠, 董雪玉, 巩文洋, 等. 饲粮粗蛋白质和粗脂肪水平对生长期貉生长性能、血清生化指标及游离氨基酸含量的影响[J]. 动物营养学报, 2018,30(12):5075-5082.
	Li M, Dong XY, Gong WY, et al. Effects of dietary crude protein and crude fat levels on growth performance, serum biochemical indexes and free amino acid content of growing raccoon dogs[J]. Acta Zoonutrimenta Sinica, 2018,30(12):5075-5082.

菌株	特性	来源
里氏木霉（Trichoderma reesei）	产生纤维素酶,分解植物粗纤维^[6]	实验室保存
毕赤酵母（Pichia pastoris）	产生单细胞蛋白^[7]	实验室保存
植物乳杆菌（Lactobacillus plantarum）	益生菌,生长力旺盛,杀死肠道有害菌^[8]	实验室保存
地衣芽孢杆菌（Bacillus licheniformis）	分泌抗菌物质,杀死其他致病菌^[9]	实验室保存
黑曲霉菌（Aspergillus niger）	淀粉酶、糖化酶以及纤维素酶,分解植物有机物^[10]	实验室保存
酿酒酵母菌（Saccharomyces cerevisiae）	积累大量蛋白质^[11]	实验室保存
枯草芽孢杆菌（Bacillus subtilis）	益生菌,生长力旺盛,杀死肠道其他有害菌^[12]	实验室保存
干酪乳杆菌（Lactobacillus casei）	益生菌,生长力旺盛,杀死肠道其他有害菌^[13]	实验室保存

菌株	特性	来源
里氏木霉（Trichoderma reesei）	产生纤维素酶,分解植物粗纤维^[6]	实验室保存
毕赤酵母（Pichia pastoris）	产生单细胞蛋白^[7]	实验室保存
植物乳杆菌（Lactobacillus plantarum）	益生菌,生长力旺盛,杀死肠道有害菌^[8]	实验室保存
地衣芽孢杆菌（Bacillus licheniformis）	分泌抗菌物质,杀死其他致病菌^[9]	实验室保存
黑曲霉菌（Aspergillus niger）	淀粉酶、糖化酶以及纤维素酶,分解植物有机物^[10]	实验室保存
酿酒酵母菌（Saccharomyces cerevisiae）	积累大量蛋白质^[11]	实验室保存
枯草芽孢杆菌（Bacillus subtilis）	益生菌,生长力旺盛,杀死肠道其他有害菌^[12]	实验室保存
干酪乳杆菌（Lactobacillus casei）	益生菌,生长力旺盛,杀死肠道其他有害菌^[13]	实验室保存