β-胡萝卜素属于多烯烃类,是水果和蔬菜中含量最高的类胡萝卜素,能够预防并减轻心脏病,白内障和癌症等疾病,市场需求量较大,利用生物合成法生产β-胡萝卜素具有较好的应用前景。胞质乙酰辅酶A是β-胡萝卜素合成的重要前体,如何提高胞质乙酰辅酶A的含量是高效合成β-胡萝卜素的关键。该研究利用米曲霉(Aspergillus oryzae)来源的ATP-柠檬酸裂解酶基因(ATP-citrate lyase,ACL)转化出发菌株ZS20,同时过表达酿酒酵母(Saccharomyces cerevisiae)自身CAB1基因,从而增加胞质乙酰辅酶A含量,β-胡萝卜素产量显著提高,菌株L5的摇瓶发酵产量达到333.3 mg/L,是出发菌株的2.1倍。在此基础上,菌株L5回补缺陷的腺嘌呤、赖氨酸、亮氨酸和色氨酸合成关键基因得到菌株L11,β-胡萝卜素摇瓶发酵产量为378.1 mg/L。经发酵罐发酵优化后,产量提高到1 152.7 mg/L。
β-carotene, a member of the polyolefin family, is the predominant carotenoid found in fruits and vegetables.It possesses the ability to prevent and alleviate various diseases, such as heart disease, cataracts, and cancer.As a result, it is highly sought after in the market due to its potential health benefits.The potential for β-carotene production through biosynthesis holds promising prospects.Significantly, cytoplasmic acetyl coenzyme A is a crucial precursor for β-carotene synthesis, highlighting the importance of increasing its content for efficient β-carotene production.In this study, the ATP-citrate lyase (ACL) from Aspergillus oryzae was introduced into the starter strain Saccharomyces cerevisiae ZS20, in which the CAB1 gene was also overexpressed, to increase the cytoplasmic acetyl-coenzyme A content.As a result, the β-carotene production significantly increased, reaching 333.3 mg/L in the flask fermentation of strain L5, which was a 2.1-fold increase compared to the starter strain.To further enhance the yield of β-carotene production, strain L5 was utilized to complement the deficient key genes responsible for adenine, lysine, leucine, and tryptophan synthesis, resulting in the development of strain L11.Initially, strain L11 demonstrated a β-carotene yield of 378.1 mg/L when cultivated in shaking flasks.Subsequently, through additional optimization of the fermentation process carried out in the fermenter, the yield was significantly enhanced to 1 152.7 mg/L.
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