Leppik R A, Park R J, Smith M G
Appl Environ Microbiol. 1982 Oct;44(4):771-6. doi: 10.1128/aem.44.4.771-776.1982.
Seventy-eight stable cultures obtained by enrichment on media containing ox bile or a single bile acid were able to utilize one or more bile acids, as well as components of ox bile, as primary carbon sources for growth. All isolates were obligate aerobes, and most (70) were typical (48) or atypical (22) Pseudomonas strains, the remainder (8) being gram-positive actinomycetes. Of six Pseudomonas isolates selected for further study, five produced predominantly acidic catabolites after growth on glycocholic acid, but the sixth, Pseudomonas sp. ATCC 31752, accumulated as the principal product a neutral steroid catabolite. Optimum growth of Pseudomonas sp. ATCC 31752 on ox bile occurred at pH 7 to 8 and from 25 to 30 degrees C. No additional nutrients were required to sustain good growth, but growth was stimulated by the addition of ammonium sulfate and yeast extract. Good growth was obtained with a bile solids content of 40 g/liter in shaken flasks. A near-theoretical yield of neutral steroid catabolites, comprising a major (greater than 50%) and three minor products, was obtained from fermentor growth of ATCC 31752 in 6.7 g of ox bile solids per liter. The possible commercial exploitation of these findings to produce steroid drug intermediates for the pharmaceutical industry is discussed.
通过在含有牛胆汁或单一胆汁酸的培养基上富集培养获得的78种稳定培养物,能够利用一种或多种胆汁酸以及牛胆汁的成分作为生长的主要碳源。所有分离株均为专性需氧菌,其中大多数(70株)为典型(48株)或非典型(22株)假单胞菌菌株,其余(8株)为革兰氏阳性放线菌。在选择用于进一步研究的6株假单胞菌分离株中,有5株在甘氨胆酸上生长后主要产生酸性分解代谢产物,但第六株,即假单胞菌属ATCC 31752,积累了一种中性类固醇分解代谢产物作为主要产物。假单胞菌属ATCC 31752在牛胆汁上的最佳生长pH值为7至8,温度为25至30摄氏度。维持良好生长不需要额外的营养物质,但添加硫酸铵和酵母提取物可刺激生长。在摇瓶中,胆汁固体含量为40 g/升时可获得良好生长。从ATCC 31752在每升6.7 g牛胆汁固体的发酵罐培养中,获得了接近理论产量的中性类固醇分解代谢产物,包括一种主要产物(大于50%)和三种次要产物。本文讨论了将这些发现用于商业开发以生产制药行业甾体药物中间体的可能性。