Malaviya Alok, Gomes James
Department of Biochemical Engineering & Biotechnology, Indian Institute of Technology Delhi, Hauz Khas, New Delhi 110016, India.
Appl Biochem Biotechnol. 2009 Aug;158(2):374-86. doi: 10.1007/s12010-008-8416-8. Epub 2009 Feb 3.
Androstenedione (AD) is a steroid intermediate used in the pharmaceutical industry for the production of several important anabolic drugs. An important route for producing AD is by the microbial transformation of sterols. Microbes capable of efficiently transforming sterols to AD are few and newer strains need to be isolated. Conventional procedures for screening and isolation are time consuming. A new procedure was used for screening and isolation of fungal microorganisms capable of biotransformation of sitosterol to androstenedione. In this procedure, Basic Alignment Search Tool (BLAST) and Position-Specific Iterative BLAST were employed to obtain a parent set of candidate microorganisms. The parent set was reduced using heuristics and constraints to obtain a manageable number of microorganisms that may be tested experimentally. For this work, screening of the entire NCBI database yielded a parent set containing 64 microorganisms. Among these, only two microorganisms, Aspergillus oryzae and Aspergillus nidulans FGSC A4, qualified to the experimental stage. Sitosterol biotransformation experiments were carried out using A. oryzae and the production of AD in culture medium was confirmed.
雄烯二酮（AD）是一种甾体中间体，在制药工业中用于生产几种重要的合成代谢药物。生产AD的一条重要途径是通过甾醇的微生物转化。能够有效地将甾醇转化为AD的微生物很少，需要分离出新的菌株。传统的筛选和分离程序耗时较长。一种新的程序被用于筛选和分离能够将谷甾醇生物转化为雄烯二酮的真菌微生物。在这个程序中，使用基本局部比对搜索工具（BLAST）和位置特异性迭代BLAST来获得候选微生物的亲本集。使用启发式方法和约束条件减少亲本集，以获得数量可控的可进行实验测试的微生物。对于这项工作，对整个NCBI数据库进行筛选得到了一个包含64种微生物的亲本集。其中，只有两种微生物，米曲霉和构巢曲霉FGSC A4，进入了实验阶段。使用米曲霉进行了谷甾醇生物转化实验，并证实了培养基中AD的产生。
