【作者】 翟红梅；

【导师】 肖冬光；

【作者基本信息】 天津科技大学 ， 发酵工程， 2004， 硕士

【摘要】 本文研究了红酵母类胡萝卜素的提取方法和发酵培养条件，利用紫外诱变的方法选育类胡萝卜素高产菌株，并对红酵母类胡萝卜素的组成进行初步分析。 通过对不同破壁方法和提取溶剂的比较得到，最佳破壁方法是酸热法，最适提取溶剂为石油醚：丙酮（1：1）混合溶液。 通过单因素实验，以及正交、均匀设计、响应面优化等统计方法优化得到发酵培养最佳培养基组成为：蔗糖40g／L，酵母粉20g／L、K₂SO₄1.0％、MgSO₄·7H₂O 0.05％、CuSO₄·5H₂O 0.0045％、VB₁0.2mg／L、VB₂ 1.0mg／L、VC 0.7mg／L、番茄汁3.0mL／L，花生油0.8mL／100mL；最佳发酵条件：培养基初始值pH6.0，装液量30mL／500mL，发酵温度28℃，摇床转速150r／min，发酵周期84h。在优化条件下生物量15.63g／L、类胡萝卜素含量941.14 μg／g及产量4.71mg／L，比在初始条件下（7.46g／L，86.73μg／g，0.647mg／L）分别提高1.09倍、9.85倍和35.14倍。 对红酵母发酵类胡萝卜素5L罐培养条件进行了研究，最佳条件为：总糖浓度80g／L，酵母粉20g／L，流加培养，糖浓度维持在10～20g／L间，pH值为6.0，DO值维持在15～25％之间。利用MATLAB软件对发酵过程进行研究，建立了菌体生长、产物形成的动力学模型，拟和模型能较好反映类胡萝卜素分批发酵过程。 利用二苯胺为筛选剂，通过紫外诱变，经筛选、发酵性能测定及遗传稳定性鉴定，选出菌株Ruv701为高产菌，摇瓶培养产量高于出发菌株42.06％，5L罐流加培养提高62.56％。 利用薄层分析及HPLC的方法，对所得类胡萝卜素进行分析，发现红酵母类胡萝卜素至少由4种色素组成，其中一种为β-胡萝卜素。更多还原

【Abstract】 Extraction method and fermentation conditions of carotenoid were studied in the paper. Rhodotorula glutins was employed in the study. The high yield strains were obtained through UV mutagenesis. At last the compositions of carotenoid were analyzed.Different methods of breaking cell wall and extracting solvent were compared. The acid-heat method and the solvent of petroleum ether and acetone (1:1) were appropriate.Through single factor experiment, orthogonal test, uniform design and Box-Burman method, the optimal fermented condition were obtained as follows: sucrose40g/L,yeast extract 20g/L, K2SO4 1.0%, MgSO4 7H2O 0.05%, CuSO4 5H2O 0.0045%, VB1 0.2 mg/L,VB2 1.0mg/L, VC 0.7 mg/L, tomato juice 3.0 ml/L, peanut oil 0.8ml/100ml, pH 6.0, medium volume 30ml (in 500ml flask) , 28 C,150rpm and culture time 84h. The cell biomass, carotenoid content and yield could reach 15.63g/L, 941.14 g/g, 14.71 mg/L respectively under the selected optimum conditions.The best conditions of 5L fermentor were as follows: the total sucrose concentration 80g/L, yeast extract 20g/L, sucrose concentration was maintained 10~20g/L, pH6.0, DO15~25%. The fermentation kinetics was studied by MATLAB. Two kinetic models were constructed which could reflect the regularity of growth, production formation in the process of batch fermentation.The start strain was treated with ultraviolet. Carotenoid synthesis inhibitor dipheny-lamine was used in selection of the mutants. Ruv701 was selected through determination of characters of mutants. The mutant has good stabilization of inheritance. The production of carotenoid was increased 42.06% and 62.56% respectively than the start strain in flask and fermentor.TLC and HPLC method were used to separate and identify Rhodotorular glutinis pigments in the text. Four components were separated and one of them could be -carotenoid; others could not be identified accurately here.更多还原