摘要
重组大肠杆菌 E.coli XL-1 Blue（pKSSE5.3）携带Ralstonia eutropha H16的 PHA聚合酶基因（phaC）和Clostridium kluyveri的4-羟基丁酸:CoA转移酶基因（orfZ）,可以利用葡萄糖和4-羟基丁酸为碳源合成均聚的聚-4-羟基丁酸[P（4HB）]。优化培养基和培养条件后,进行了补料分批培养。结果表明,经68h左右培养,E.coli XL-1 Blue（pKSSE5.3）的发酵液中菌体干重达13g/L,P（4HB）的密度达5g/L,P（4HB）百分含量为36％。从收获的冻干细胞中提纯得到40g均聚的P（4HB）,为进一步分析检测P（4HB）生物、理化、加工特性及其应用价值成为可能。
A recombinant E. coli XL1-Blue (pKSSE5.3) habouring PHA sythase gene (phaC) of Ralstonia eu-tropha H16 and 4-hydroxybutyric acid: CoA transferase gene (orfZ) of Clostridium kluyveri was used to produce poly (4-hydroxybutyric acid), P (4HB), homopoiyester in M9 salts medium with glucose and 4-hydroxybutyric acid as carbon sources. Optimization of medium type and cultivation conditions and fed-batch culture were carried out. The results showed that the final cell dry weight, P (4HB) concentration and P (4HB) content were 13 g/L, 5g/L, and 36%, respectively in a 27L stirred and aerated fermentor after 68 hours of fed-batch culture. A large amount of P (4HB) was isolated by extraction of lyophilized cells with chloroform and make it possible to further examine the material properties and applications of P (4HB) homopoiyester from recombinant E. coli.
出处
《微生物学通报》
CAS
CSCD
北大核心
2002年第1期10-14,共5页
Microbiology China
基金
德国教育科研部(BMBF)部长基金资助项目
