粉被虫草静置发酵产N^6-(2-羟乙基)腺苷培养基组分及前体物的筛选

Screening of culture medium constituents and additives for producing N^6-(2-hydroxyethyl)-adenosine by Cordyceps pruinosa strain cp incubated statically

在静置培养条件下,对粉被虫草cp菌株产N6‐(2‐羟乙基)腺苷(简称HEA)的培养基及其组分进行优化。采用单因素实验方法,以HPLC法对cp菌丝体中HEA进行检测,以HEA的产量为指标,结果表明查氏培养基有利于cp菌株产HEA,其产量是沙氏和PD培养基的3.7倍和4.48倍。以查氏培养基为基础培养基,进行碳、氮源的筛选中,最有利于cp菌株产HEA的是蔗糖和硝酸钠;在无机盐的筛选中,K2HPO4是促进cp菌株累积HEA最主要的无机盐,其HEA产量较CK增长了5 822.37%(提高了58.2倍);在查氏培养基上添加不同前体物和氨基酸,发现其结构类似物腺苷、次黄嘌呤和腺嘌呤都有促进cp菌株累积HEA的能力,其中腺苷和次黄嘌呤能提高其产量60%以上;而组氨酸是最有利于cp菌株累积HEA的氨基酸,产量达到(45.56±2.8)mg/L,较CK提高HEA产量251.68%,其次是L‐谷氨酸增产184.19%。

Under static culture conditions, the single factor experiments were applied to optimize the N6-（2-hydroxyethyl）-adenosine （HEA） fermentation medium by Cordyceps pruinosa cp. The detection of HEA was done by HPLC method. The results showed that Czapek medium was the best one for the strains cp to produce HEA, its yield was 3.7 times and 4.48 times as high as that of Sabouraud and PD medium respectively. When the basal medium was Czapek medium, the most conducive carbon and nitrogen sources were sucrose and sodium nitrate respectively for the strains cp producing HEA. The promoting factor of inorganic salts for the cp strains accumulated HEA was K2HPO4 which could increase the yield of HEA by 5 822.37% （58.2 times） as compared with CK’s. Additions of adenosine, hypoxanthine and adenine considerably increased the HEA production, especially the addition of adenosine and hypoxanthine which increasing the HEA production by 60% as compared with the control. Histidine is the most beneficial amino acid for the cp strains to produce HEA and the yield reached （45.56±2.8）mg/L which increased 251.68% as compared with CK’s. Secondarily, L-glutamic acid increased the yield by 184.19%.