基于响应面法优化滑菇液体菌种发酵培养基配方  

Optimization of liquid fermentation medium for Pholiota microspora based on response surface methodology

作　　者：陈珣[1] 肇莹[1] 龚娜[1] 刘国丽[1] 马晓颖[1] 张敏[1] 肖军[1] CHEN Xun;ZHAO Ying;GONG Na;LIU Guo-li;MA Xiao-ying;ZHANG Min;XIAO Jun(Institute of Edible Fungi,Liaoning Academy of Agricultural Science,Shenyang,Liaoning 110161,China)

机构地区：[1]辽宁省农业科学院食用菌研究所,辽宁沈阳110161

出　　处：《南方农业学报》2024年第12期3625-3635,共11页Journal of Southern Agriculture

基　　金：国家食用菌产业技术体系专项(CARS-20);辽宁省“揭榜挂帅”科技计划项目(2021JH1/10400035);辽宁省应用基础研究计划项目(2022JH2/101300160);沈阳市种业创新专项(22-318-2-20)。

摘　　要：【目的】基于响应面法探究不同碳源、氮源、无机盐种类及浓度对滑菇液体菌种生产的影响,为寻找最适滑菇菌种生长的配方,缩短滑菇生产周期,降低生产成本及促进滑菇规模化生产提供理论依据。【方法】以滑菇菌株滑99为研究对象,基础培养基为空白对照(CK),选取5种碳源(玉米粉、蔗糖、麦芽糖、可溶性淀粉和甘露醇),5种氮源(黄豆粉、牛肉膏、酵母浸粉、蛋白胨和酒石酸钠)及5种无机盐(硫酸镁、磷酸二氢钾、氯化钠、硫酸锰和氯化钙)作为配方筛选因子,利用单因素试验筛选最适碳源、氮源和无机盐种类,设不同浓度梯度筛选最佳碳源、氮源、无机盐的浓度,采用响应面法优化滑菇液体菌种发酵培养基配方并进行验证。【结果】玉米粉作为碳源时,滑菇菌种的菌球数(70个)和菌丝生长速率(1.471 cm/d)均显著高于CK和其他碳源(P<0.05,下同);黄豆粉作为氮源时,菌丝生物量最高(6.200 g/L);硫酸镁作为无机盐时,滑菇菌丝生长速率最高(0.638 cm/d),菌丝生物量最高(3.102 g/L)。玉米粉、黄豆粉和硫酸镁分别为滑菇液体菌种培养的最佳碳源、氮源和无机盐;单因素试验结果显示,最适滑菇菌种生长的玉米粉、黄豆粉和硫酸镁浓度分别为30、20和3 g/L。以玉米粉(A)、黄豆粉(B)、硫酸镁(C)为因变量,以菌丝生物量作为响应值(Y),建立二次多项回归方程:Y=15.448+1.531A+1.739B-0.063C+0.312AB+0.223AC-0.914BC-0.176A^(2)-3.139B^(2)-1.046C^(2),根据F值,确定各因素对滑菇菌种菌丝生物量影响大小的排序为黄豆粉(B)>玉米粉(A)>硫酸镁(C);黄豆粉与硫酸镁(BC)之间的交互作用明显。配方验证试验结果显示,菌丝体生物量实测值为17.223 g/L,与理论值17.143 g/L基本相符。【结论】利用响应面法优化滑菇液体菌种发酵培养基配方为玉米粉30 g/L+黄豆粉16.67 g/L+硫酸镁2.93 g/L,且该优化配方有效可行。【Objective】Based on the response surface method,the effects of different carbon sources,nitrogen sources,inorganic salt types and concentrations on the production of liquid strain of Pholiota microspora were investigated,which provided theoretical basis for finding the most suitable formula for the growth of P.microspora,shortening the production cycle,reducing the production cost and promoting the large-scale production of P.microspora.【Method】The P.microspora strain Hua 99 was selected as the study object,and the basic medium was the blank control(CK).Five carbon sources(corn flour,sucrose,maltose,soluble starch and mannitol),five nitrogen sources(soybean flour,beef extract,yeast infusion powder,peptone and sodium tartrate)and five inorganic salts(MgSO_(4),KH_(2)PO_(4),NaCl,MnSO_(4)and CaCl_(2))were used as formula screening factors.Single factor test was used to screen the most suitable carbon sources,nitrogen sources and inorganic salts.The concentration gradient test was set up to screen the best carbon source,nitrogen source and inorganic salt concentrations,and the response surface method was used to optimize the fermentation formula of liquid strain of P.microspora and verify it.【Result】When corn flour was used as carbon source,the globule count(70)and the mycelium growth rate(1.471 cm/d)were significantly higher than CK and other carbon sources(P<0.05,the same below).When soybean flour was used as nitrogen source,the mycelium biomass was the highest(6.200 g/L).When MgSO_(4)was used as inorganic salt,the mycelium growth rate was the fastest(0.638 cm/d),and the mycelium biomass was the highest(3.102 g/L).Corn flour,soybean flour and MgSO_(4)were the best carbon source,nitrogen source and inorganic salt for liquid strain of P.microspora respectively.The results of single factor test showed that the optimal concentrations of corn flour,soybean flour and MgSO_(4)were 30,20 and 3 g/L.With corn flour(A),soybean powder(B)and MgSO_(4)(C)as dependent variables and mycelium biomass as response value(Y),

关 键 词：滑菇 液体发酵 响应面法 菌丝生物量 

分 类 号：S646.16[农业科学—蔬菜学]