产辅酶Q10的类球红细菌原生质体制备和再生研究

 辅酶Q₁₀(CoQ₁₀)是人类细胞自身产生的内源性辅酶因子,是一类天然抗氧化剂,目前已广泛应用于医药、食品、化妆品等诸多领域.基因组改组技术的核心内容为原生质体融合,为了提高基因组改组的效率,提高辅酶Q₁₀的产量,本研究对产辅酶Q₁₀的类球红细菌原生质体制备及再生条件进行了优化,通过生长曲线、正交试验等确定了原生质体制备及再生的适宜条件：溶菌酶浓度1mg/mL、作用温度37℃、作用时间1h以及再生培养基蔗糖浓度10%.在此条件下,类球红细菌原生质体形成率可达到96.1%,再生率可达到28.8%.这为进一步对该菌的基因组改组研究奠定了基础.

Protoplast Preparation and Regeneration of Coenzyme Q10 Producing Strain Rhodobacter sphaeroides

Coenzyme Q₁₀ (CoQ₁₀ ) is an endogenous enzyme cofactor that may be found in every cell in the human body and an important natural antioxidant widely used in many fields, such as medicine, food and cosmetics. The protoplast fusion is an important process in the genome shuffling. In order to improve the efficiency of the genome shuffling and the CoQ₁₀ production, this paper studies the conditions of the protoplast preparation and the regeneration of CoQ₁₀ to produce the strain Rhodobacter sphaeroides. The parameters of the proper protoplast formation and the regeneration were determined through the growth curve and the orthogonal experiment: the lysozyme concentration is 1mg/ml, the enzymatic temperature is 37℃; the enzymatic time is 1h and the sucrose concentration is 10%, and under these conditions, the protoplast formation rate is 96.1% and the protoplast regeneration rate reaches 28.8%. Therefore, the optimized conditions of the protoplast preparation and the regeneration for the genome shuffling of Rhodobacter sphaeroides are obtained.