摘要
【目的】研究菌株CY-2012菌丝体内三萜类物质的产生条件,以期得到最高量的三萜类物质。【方法】以28℃,150 r·min^(-1)作为初始培养条件,用基础培养基对采自长白山的栗黑层孔菌菌株CY-2012进行培养,然后采用2项单因子试验、1次4因素3水平的正交试验,对该菌株产三萜类物质的液体培养基成分进行优化;在此基础上,应用异丙醇和硫酸铵双水相萃取技术提取菌丝体内的三萜,采用2项单因子试验、1次3因素3水平的正交试验,对培养基成分优化后获得的菌丝体进行三萜类物质提取工艺研究。【结果】菌株CY-2012产三萜最优的培养基成分组合是:去皮马铃薯200 g·L^(-1)、葡萄糖25.0 g·L^(-1)、乳糖3.0 g·L^(-1)、酵母15.0 g·L^(-1)、MgSO_4·7H_2O 0.5 g·L^(-1)、KH_2PO_40.5 g·L^(-1)、NH_4NO_30.3 g·L^(-1)、维生素B10.5 g·L^(-1),在28℃、150 r·min-1条件下培养10天菌丝干质量达0.769 g·(100 m L)-1,其中三萜类物质产量高达14.529 mg·(100 m L)-1,分别是初始培养条件下的9.96和35.52倍。双水相萃取技术最佳的提取工艺为异丙醇体积分数40%,硫酸铵质量浓度0.18 g·mL^(-1),室温浸提2 h,超声时间35 min,得到的三萜提取率为3.65%。【结论】通过优化培养基成分可以较大幅度地提高三萜的产量,优化异丙醇和硫酸铵双水相萃取提取工艺可以提高三萜提取率。
【Objective 】 This study investigated the production of intracellular triterpenoids in the fungus strain Melanoporia castanea CY-2012 in order to obtain the highest amount of triterpenoids. 【Method 】The strain CY-2012,collected in Changbai Mountains, was first cultured under 28 ℃,150 r·min-1. Subsequently, the submerged fermentation medium components were optimized through 2 single-factor experiments and one orthogonal experiment of fourvariables with three-levels. Furthermore,isopropyl alcohol-ammonium sulfate aqueous two-phase systems were used to extract intracellular triterpenes of mycelia which were obtained under the optimal medium condition,and the extraction technique was optimized through 2 single-factor experiments and an orthogonal experiment of three-variables with threelevels. 【Result】The optimal medium components were composed of peeled potatoes 200 g·L-1,glucose 25. 0 g·L-1,lactose 3. 0 g·L-1,yeast extract 15. 0 g·L-1,Mg SO40. 5 g·L-1,KH2PO40. 5 g·L-1,NH4NO30. 3 g·L-1,and VB10.5 g·L-1. After the fhe fungus was incubated for ten days under 28 ℃,the mycelium dry weight reached 0. 769 g·（ 100 mL-1） which was 9. 96 times greater than that produced at the initial conditions,and the yield of intracellular triterpenoids reached 14. 529 mg·（ 100 mL-1） which was 35. 52 times greater than that produced at the initial conditions.The best extraction process with the two-phase extracting agent was under 40% isopropyl alcohol,and 0. 18 g·mL-1ammonium sulfate in water,the mycelia were steeped for 2 hours at room temperature,and the extract was treated with ultrasonic for 35 min,and the triterpenoids extraction rate was 3. 65%. 【Conclusion 】Optimized medium components could substantially boost the yield of total triterpenoids,and optimized isopropyl alcohol-ammonium sulfate aqueous twophase systems could enhance the triterpenoids extraction rate.
出处
《林业科学》
EI
CAS
CSCD
北大核心
2016年第9期147-153,共7页
Scientia Silvae Sinicae
基金
国家自然科学基金项目(30671700)
黑龙江省自然科学基金项目(C2016006)
关键词
栗黑层孔菌
三萜化合物
优化
双水相体系
提取工艺
Melanoporia castanea
triterpenoids
optimization
aqueous two-phase systems
extraction technique
