响应面法优化产二氢大豆苷元菌株发酵培养基

doi:10.6048/j.issn.1001-4330.2018.01.016

摘要/Abstract

摘要： 【目的】 研究采用单因素试验设计和响应面分析法对产二氢大豆苷元菌株发酵培养基的主要营养成分进行优化。【方法】 以脑浸心肉汤培养基(BHI)为基础,通过单因素试验确定主要因素(碳源、氮源、生长因子和无机盐)及其适宜的浓度范围,利用Box-Behnken中心组合试验设计,采用响应面法进行回归分析,确定培养基中的最佳组成成分。【结果】 通过单因素试验确定了葡萄糖、蛋白胨、VB₁和KH₂PO₄能够提高二氢大豆苷元的产量,响应面分析法进一步确定在以BHI为基础的条件下,葡萄糖的补充量为10.22 g/L、蛋白胨为6.00 g/L、VB₁为0.49 g/L、KH₂PO₄为0.82 g/L。验证试验显示在培养基优化后的条件下,DHD的产生量为0.27 μg/mL,与预测值0.28 μg/mL接近。【结论】 利用单因素试验和响应面法,分析明确了产二氢大豆苷元菌株发酵培养基最佳成分为38 g/L的BHI、10.22 g/L的葡萄糖、6.00 g/L的蛋白胨、0.49 g/L的VB₁、0.82 g/L的KH₂PO₄。

关键词: 二氢大豆苷元; 响应面法; BHI培养基

Abstract: 【Objective】 he aim of this experiment is to optimize the main nutrient components of the fermentation medium of dihydrodaidzein producing strain by single factor test design and response surface methodology.【Method】 On the basic of BHI medium, the main factors (carbon source, nitrogen source, growth factor and inorganic salt) and the appropriate range of concentration were determined by single factor test. The best composition in the medium was analyzed by the Box-Behnken central composite experiment and response surface methodology.【Result】 The significant factors such as glucose, peptone,VB₁ and KH₂PO₄ were determined by the single factor experiment and response surface methodology analysis was determined that the supplemental doses to the strain medium were 10.22 g/L glucose, 6.00 g/L peptone, 0.49 g/L VB₁ and 0.82 g/L KH₂PO₄ on the basic of BHI medium. Validation experiments showed that the DHD concentration was 0.27 μg/mL, which was close to the predictive value 0.28 μg/mL.【Conclusion】 The results from the single factor experiment and response surface methodology showed that the optimal ingredients of medium for DHD production were 38 g/L BHI,10.22 g/L glucose,6.00 g/L peptone,0.49 g/L VB₁ and 0.82 g/L KH₂PO₄.

Key words: dihydrodaidzein; response surface methodology; BHI medium

中图分类号:

S188

谢景龙,李晓斌,赵芳,王晨晨,杨开伦. 响应面法优化产二氢大豆苷元菌株发酵培养基[J]. 新疆农业科学, 2018, 55(1): 143-155.

XIE Jing-long, LI Xiao-bin, ZHAO Fang, WANG Chen-chen, YANG Kai-lun. Optimization of Fermentation Medium Components for Dihydrodaidzein Producing Strain Using RSM[J]. Xinjiang Agricultural Sciences, 2018, 55(1): 143-155.

参考文献 28

[1]	汪秋安,周冰,单杨.天然黄酮类化合物的抗氧化活性和提取技术研究进展[J].化工生产与技术,2004,11(5):29-32.WANG Qiu-an, ZHOU Bing, SHANG Yang. (2004). Research progress on antioxidant activity and extraction technology of natural flavonoids [J]. Chemical Production and Technology,11(5):29-32.(in Chinese)
[2]	徐梦蕾,高宇,刘静波,等.基于抗氧化的大豆异黄酮对PC12细胞的神经保护作用[J].吉林大学学报(工学版),2017,47(1):332-336. XU Meng-lei, GAO Yu, LIU Jing-bo, et al.(2017). Neuroprotective effects of soy isoflavones on PC12 cells based on antioxidation [J]. Journal of Jilin University(Engineering and Technology Edition) , 47(1):332-336.(in Chinese)
[3]	Vitale, D. C., Piazza, C., Melilli, B., Drago, F., & Salomone, S. (2013). Isoflavones: estrogenic activity, biological effect and bioavailability. European Journal of Drug Metabolism & Pharmacokinetics, 38(1): 15-25.
[4]	Liu, W. K., Cheung, F. W., Liu, B. P., Li, C., Ye, W., & Che, C. T. (2008). Involvement of p21 and fasl in induction of cell cycle arrest and apoptosis by neochamaejasmin a in human prostate lncap cancer cells. ,71(5):842-846.
[5]	Campbell, C. T., Aich, U., Weier, C. A., Wang, J. J., Choi, S. S., & Wen, M. M., et al. (2008). Targeting pro-invasive oncogenes with short chain fatty acid-hexosamine analogs inhibits the mobility of metastatic mda-mb-231 breast cancer cells. Journal of Medicinal Chemistry, 51(24): 8,135-8,147.
[6]	Levis, S., Strickman-Stein, N., Doerge, D. R., & Krischer, J. (2010). Design and baseline characteristics of the soy phytoestrogens as replacement estrogen (spare) study - a clinical trial of the effects of soy isoflavones in menopausal women. Contemporary Clinical Trials, 31(4): 293-302.
[7]	Giordano, E., Dávalos, A., Crespo, M. C., Tomécarneiro, J., Gómezcoronado, D., & Visioli, F. (2015). Soy isoflavones in nutritionally relevant amounts have varied nutrigenomic effects on adipose tissue. Molecules, 20(2): 2,310-2,322.
[8]	Tamura, M., Hori, S., Nakagawa, H., Katada, K., Kamada, K., & Uchiyama, K., et al. (2015). Relationships among fecal daidzein metabolites, dietary habit and bmi in healthy volunteers: a preliminary study. Bioscience & Microflora, 34(3): 59-65.
[9]	Setchell, K. D. R., & Cole, S. J. (2006). Method of defining equol-producer status and its frequency among vegetarians. Journal of Nutrition, 136(8): 2,188-2,193.
[10]	Morton, M. S., Arisaka, O., Miyake, N., Morgan, L. D., & Evans, B. A. J. (2002). Phytoestrogen concentrations in serum from japanese men and women over forty years of age. Journal of Nutrition, 132(10): 3,168-3,171.
[11]	Nielsen, I. L., & Williamson, G. (2007). Review of the factors affecting bioavailability of soy isoflavones in humans. Nutrition & Cancer, 57(1): 1-10.
[12]	Barnes, S. (2010). The biochemistry, chemistry and physiology of the isoflavones in soybeans and their food products. Lymphatic Research & Biology, 8(1): 89-98.
[13]	Lampe, J. W. (2009). Is equol the key to the efficacy of soy foods. American Journal of Clinical Nutrition, 89(5): 1,664S-1,667S.
[14]	王宇,尹业师,朱立颖,等.人粪样中雌马酚含量调查及其与菌群结构相关性研究[J].中国微生态学杂志, 2013, 25(7):757-761.WANG Yu, YIN Ye-shi, ZHU Li-ying, et al. (2013).The level of equol its relationship with the bacterial community in human fecal samples[J]. Chinese Journal of Microecology, 25(7):757-761. (in Chinese)
[15]	张逊.猪肠道大豆苷原降解菌分离及其性质研究[D].南京: 南京农业大学硕士学位论文,2007.ZHANG Xun. (2007). Isolation and characterizing od daidzein-degradion bacteria from the intestinal tract of pig [D]. Master Dissertation. Nanjing Agricultural University, Nanjing.(in Chinese)
[16]	李笑梅,贾尧.素食者产雌马酚肠道菌生长条件优化[J].食品科学, 2014, 35(23):199-203.LI Xiao-mei, JIA Yao. (2014).Growth Characteristics of Equol-Producing Bacterial Strains from the intestinal of Vegetarians and Optimization of Culture Conditions for Enhanced Equol Production [J]. Food Science, 35(23):199-203. (in Chinese)
[17]	郝学财,余晓斌,刘志钰,等.响应面方法在优化微生物培养基中的应用[J].食品研究与开发,2006,27(1):38-41.HAO Xue-cai, YU Xiao-bin, LIU Zhi-yu, et al. (2006). The application of response surface method in optimizing microbial medium [J]. Food Research and Development, 27(1):38-41.(in Chinese)
[18]	Rowland, I. R., Wiseman, H., Sanders, T. A., Adlercreutz, H., & Bowey, E. A. (2000). Interindividual variation in metabolism of soy isoflavones and lignans: influence of habitual diet on equol production by the gut microflora. Nutrition & Cancer, 36(1): 27-32.
[19]	于卓腾.大豆黄酮对仔猪肠道微生物的影响和雌马酚产生菌的分离及其特性的研究[D].南京:南京农业大学博士学位论文,2007.YU Zhuo-teng. (2007). Effects of daidzein on the gastrointestinal microbial community of Piglets and isolation and study characterisation of equol-producing bacteria [D]. PhD Dissertation. Nanjing Agricultural University, Nanjing. (in Chinese)
[20]	郭远洋.生物催化大豆苷元产S-雌马酚菌株筛选及其转化特性研究[D].厦门:华侨大学硕士学位论文,2012.GUO Yuan-yang. (2012). Study on the screen of bacteria strain for converting daidzein to S-equol and the characteristics of its conversion [D]. Master Dissertation. Huaqiao University, Xiameng.(in Chinese)
[21]	杨立杰,张丽莉,李东坡,等.硝化抑制剂和秸秆对潮棕壤碳氮转化和微生物群落特征的短期影响[J].中国土壤与肥料,2017,(1):86-91.YANG Li-jie, ZHANG Li-li, LI Dong-po.et al. (2017).Short-term effects of nitrification inhibitor and straw on aquic brown soil carbon and nitrogen mineralization and microbial community structure [J]. Soil and Fertilizer Sciences, (1):86-91. (in Chinese)
[22]	邵杰,罗建光,曾晓雄.液体培养的桑黄胞外多糖发酵培养基成分的优化[J].食品科学,2012,33(3):121-125.SHAO Jie, LUO Jian-guang, ZENG Xiao-xiong. (2012).Optimization of Cultivation Conditions for Extracellular Polysaccharide Production by P. baumii Pilá [J]. Food Science, 33(3):121-125.(in Chinese)
[23]	王镜岩,朱圣康,徐长法.生物化学[M].第三版(上册),北京:高等教育出版社,2002:442.WANG Jing-yan, ZHU Sheng-kang, XU Chang-fa.(2002). Biochemistry[M].Beijing: Higher Education Press: 442.(in Chinese)
[24]	郝志敏,季昀,潘晓花,等.硫胺素(维生素B1)对体外培养瘤胃微生物发酵的影响[J].中国饲料添加剂,2012,(2):20-24.HAO Zhi-min, JI Yun, PAN Xiao-hua. (2012). Effects of Thiamin Supplement on the Rumen Microbial Fermentation in Vitro [J]. China Feed Additive, (2):20-24.(in Chinese)
[25]	郝志敏,王洪荣,潘晓花,等.高精料日粮下硫胺素对体外培养荷斯坦牛瘤胃微生物消化代谢的影响[J].中国奶牛,2011,(18):8-12.HAO Zhi-min,WANG Hong-rong, PAN Xiao-hua, et al.(2011).Effects of Thiamin on the Rumen Microbial Fermentation Under the Condition of High Concentrate Diet in Vitro [J]. China Dairy Cattle, (18):8-12.(in Chinese)
[26]	康远军.高耐性鲁氏酵母高密度发酵研究[D].武汉:湖北工业大学硕士学位论文,2015.KANG Yuan-jun.(2015). High-cell-density cultivation of a salt-tolerant yeast Zygosaccharomyces rouxii[D]. Master Dissertation. Hubei University of Technology, Wuhan.(in Chinese)
[27]	慕运动.响应面方法及其在食品工业中的应用[J].郑州工程学院学报,2001,(3):91-94.MU Yun-dong. (2001).Response surface methodology and its application in food industry [J]. Journal of Zhengzhou Institute of Technology,(3): 91-94.(in Chinese)
[28]	姜雪,刘红玉,于鹏.响应面法优化产雌马酚菌株发酵培养基[J].食品工业,2014,35(7):67-71.JIANG Xue, LIU Hong-yu, YU Peng. (2014).Optimization of Fermentation Medium Components for Equol Production Using Response Surface Methodology [J]. Food Industry, 35(7):67-71. (in Chinese)