高产类胡萝卜素的蛹虫草液体培养基优化及其提取工艺研究

殷林; 林俊芳; 钱璟; 叶志伟; 罗诗华; 郭丽琼; 云帆

doi:10.13386/j.issn1002-0306.2017.18.016

高产类胡萝卜素的蛹虫草液体培养基优化及其提取工艺研究

Optimization of the liquid culture medium composition for high carotenoid production by Cordyceps militaris and the investigation on the improvement of carotenoid extraction

摘要

摘要: 以类胡萝卜素含量为目标,采用谷物类为培养基对蛹虫草CM-03、CM-04和CM-11三株菌株进行了液体发酵培养基的优化,结果表明:三株菌株的最优培养基配方分别为CM-03:1.4 g·L^-1小麦、2.8 g·L^-1玉米、0.3 g·L^-1大豆、0.1 g·L^-1MgSO₄·7H₂O、0.1 g·L^-1KH₂PO₄、0.05 mg·L^-1VB₁、pH自然,类胡萝卜素含量为（694.8±15.4）μg/g;CM-04:1.2 g·L^-1大米、0.6 g·L^-1玉米、0.3 g·L^-1大豆,0.1 g·L^-1MgSO₄·7H₂O、0.1 g·L^-1KH₂PO₄、0.05 mg·L^-1VB₁、pH自然,类胡萝卜素含量为（708.5±25.8）μg/g;CM-11:1.7 g·L^-1小麦、2.6 g·L^-1玉米、0.3 g·L^-1大豆,0.1 g·L^-1MgSO₄·7H₂O、0.1 g·L^-1KH₂PO₄、0.05 mg·L^-1VB₁、pH自然,类胡萝卜素含量为（700.5±19.1）μg/g。同时,使用乙醇提取法对类胡萝卜素提取工艺进行了优化,结果表明乙醇提取法的最佳工艺条件为:采用石英砂研磨破壁,乙醇浓度60%、料液比1∶40,水浴提取温度80℃,提取级数2级,提取率达到92.8%。

Abstract: The cereal-based medium for liquid fermentation was optimized according to the carotenoids production of three strains of Cordyceps militaris CM-03, CM-04 and CM-11.The media components of this work contained several kinds of grains such as wheat ( W) , corn ( C) , bean ( B) , rice ( R) , bran ( Br) and inorganic salts like MgSO₄·7H₂O ( Mg) , KH₂PO₄ ( KHP) and vitamin B₁ ( VB₁) .The results showed that the best culture medium formula for the carotenoids accumulation by Cordyceps militaris CM-03 consisted of W ( 1.4 g·L^-1) , C ( 2.8 g·L^-1) , B ( 0.3 g·L^-1) , Mg ( 0.1 g·L^-1) , KHP ( 0.1 g·L^-1) and VB₁ ( 0.05 mg·L^-1) , with natural pH, with the carotenoids yield of ( 694.8 ± 15.4) μg/g.For Cordyceps militaris CM-04, its highest carotenoids output was ( 708.5 ± 25.8) μg/g under the formula consisted of R ( 1.2 g·L^-1) , C ( 0.6 g·L^-1) , B ( 0.3 g·L^-1) , Mg ( 0.1 g·L^-1) , KHP ( 0.1 g·L^-1) , VB₁ ( 0.05 mg·L^-1) with natural p H.The highest carotenoids output of Cordyceps militaris CM-11 was ( 700.5 ± 19.1) μg/g with natural p H, the formula formed by W ( 1.7 g·L^-1) , C ( 2.6 g·L^-1) , B ( 0.3 g·L^-1) , Mg ( 0.1 g·L^-1) , KHP ( 0.1 g·L^-1) and VB1 ( 0.05 mg·L^-1) .The extraction process with ethanol was also optimized in this work, and the results showed that the optimal extraction condition was as follows: grinding with quartz sand, ethanol concentration 60%, material to liquid ratio 1∶ 40, water bath temperature 80 ℃, extraction level 2, and extraction rate of92.8%.