Please wait a minute...
浙江大学学报（农业与生命科学版）
浙江大学学报(农业与生命科学版)  2013, Vol. 39 Issue (5): 481-488    DOI: 10.3785/j.issn.1008-9209.2013.02.081
生物科学与技术     
梨形马勃产胞外多糖发酵条件的响应面设计法优化
马小魁1*, 江华1, 李峻志2, 陈勇3， 何晓晶1
（1.陕西师范大学生命科学学院，药用资源与天然药物化学教育部重点实验室/西北濒危药材资源开发国家工程实验室，西安710100；2.陕西省微生物研究所，西安710043；3.陕西省产品质量监督检验所，西安710006）

Optimization of medium components for the production of exopolysaccharide by Lycoperdon pyriforme Schaeff.: Pers. using response surface methodology
MA Xiaokui1*, JIANG Hua1, LI Junzhi2, CHEN Yong3，HE Xiaojing1
(1. Key Laboratory of Medicinal Resources and Natural Pharmaceutical Chemistry, Ministry of Education / National Engineering Laboratory for Resource Developing of Endangered Chinese Crude Drugs in Northwest of China，College of Life Science, Shaanxi Normal University, Xi’an 710100, China; 2. Microbiology Institute of Shaanxi, Xi’an 710043, China; 3. Shaanxi Products Quality Supervision and Inspection Institute, Xi’an 710006, China)
 全文: PDF(2216 KB)   HTML ( ) 
摘要： 为优化梨形马勃产胞外多糖的培养基组成，运用响应面设计法对梨形马勃产胞外多糖的发酵条件进行优化研究。首先利用Minitab 16.0分析软件的Plackett-Burman设计模块对影响胞外多糖产量的培养基组分进行设计筛选，进一步结合单因素实验结果确定筛选出具有显著效应的3个因素：麦芽糖、维生素B1（VB1）和谷氨酸；然后通过该软件中的Box-Behnken实验设计模块对此3个因素进行优化，并对实验数据进行分析统计优化，结合基本培养基中其他成分含量而得到最高胞外多糖的最佳培养基组成：麦芽糖6.11%，谷氨酸0.70%，VB1  2.8×10-4%，酵母粉0.50%，KH2PO4 0.15%，MgSO4·7H2O 0.10%，初始pH值5.5。在此优化培养条件下进行验证实验，得到梨形马勃胞外多糖产量为（3.71±0.16） g/L，比采用基本培养基的多糖产量增加了4.6倍。  
Abstract: Lycoperdon pyriforme Schaeff.: Pers. is one of medicinal macro-fungi, which has become an attractive topic as a functional food and as a source for the development of drugs and nutraceuticals. Pharmaceutically active compounds including polysaccharide, steroids, polysaccharides-peptides, nucleosides and triterpenoids were reported to be found in its fruiting body, in which polysaccharides isolated from its fruiting body were  reported to possess various important biological functions such as antitumor activity, immunomodulation and antioxidant activity with low toxicity and good safety profile. However, it is known that the time required for obtaining either naturally harvested or farmed fruiting bodies of most medicinal fungal species is relatively long, while its product quality is also difficult to control in mushroom farming. In addition, even mushroom farming techniques for L. pyriforme. have not been developed adequately to meet the current and promising market for bio-based antioxidants. The submerged culture is thought as an alternative approach to produce a large amount of mycelia biomass and polysaccharides by medicinal fungi. Currently, although many studies about exopolysaccharides (EPS) bioproduction by macro-fungi have been reported, there is a little information available in literature about the optimization of medium components for enhanced EPS production by L. pyriforme. in submerged culture to date.
The objectives of this study were to screen the factors significantly  affecting EPS production by L.pyriforme. and subsequently to determine the optimal medium components for enhanced EPS yield through response surface methodology (RSM), which is a collection of statistical techniques for experimental design, model developing, evaluation factors and searching for optimum conditions, and which consists of a group of mathematical and statistical procedures. The concentration of EPS was measured by phenol-sulfuric acid method. For the optimization of medium components for enhanced EPS production, RSM was subsequently applied in this study. At  last, the validation tests were performed under the optimal conditions at shake flask scale.
Firstly, Plackett-Burman experimental design (PB) was applied to implore the relative effect of several medium components on EPS yield and vitamin B1 (VB1) was determined to affect EPS yield significantly. And combining the results of one-factor-one-time method (data not shown), three factors, namely, maltose, glutamic acid and VB1 were  further optimized  for enhanced EPS yield using Box-Behnken design (BBD). A total of 15 experimental runs (with three factors at three levels) were conducted to determine the optimum combination of these medium components by ANOVA analysis of experimental data. A second-order polynomial models with EPS yield as response variable was then obtained and confirmed to adequately represent the actual relationship between the independent variables and responses through validation tests. The optimal medium compositions were acquired as follows: maltose 6.11%, glutamic acid 0.70%, VB1 2.8×10-4%, yeast extract 0.50%, KH2PO4 0.15%, MgSO4·7H2O 0.10%, initial pH 5.5. The result of validation tests in triplicate was
(3.71±0.16) g/L under the above optimum culture conditions, and in which EPS yield was enhanced by 4.6 times compared with that produced in the basal medium in which (0.82±0.09) g/L was just obtained. In conclusion, RSM procedure is convenient, feasible and highly efficient for the optimization of culture conditions for enhanced EPS  production by fungi.  
出版日期: 2013-09-20
CLC:  S 182  
基金资助:

陕西省科技攻关资助项目(2009K1103)；陕西省科学院资助项目(201303)；西安市科技计划项目(694108);大学生创新训练计划项目(201210781116)
服务　　
把本文推荐给朋友
加入引用管理器
E-mail Alert
RSS
作者相关文章　　
马小魁1*
江华1
李峻志2
陈勇3
何晓晶1

引用本文:

马小魁1*, 江华1, 李峻志2, 陈勇3， 何晓晶1. 梨形马勃产胞外多糖发酵条件的响应面设计法优化[J]. 浙江大学学报(农业与生命科学版), 2013, 39(5): 481-488.

MA Xiaokui1*, JIANG Hua1, LI Junzhi2, CHEN Yong3，HE Xiaojing1.
Optimization of medium components for the production of exopolysaccharide by Lycoperdon pyriforme Schaeff.: Pers. using response surface methodology. Journal of Zhejiang University (Agriculture and Life Sciences), 2013, 39(5): 481-488.

链接本文:

http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2013.02.081    或    http://www.zjujournals.com/agr/CN/Y2013/V39/I5/481

No related articles found!