Production of natural indirubin from indican using non-recombinant Escherichia coli

• Published in: Bioresource technology Vol. 102; no. 19; pp. 9193 - 9198
• Main Authors: Lee, Jin-Young, Shin, Youn-Sook, Shin, Hyun-Jae, Kim, Geun-Joong
• Format: Journal Article
• Language: English
• Published: Kidlington Elsevier Ltd 01.10.2011; Elsevier
• Subjects: Biological and medical sciences; Bioreactors; Biotechnology; Biotechnology - methods; Catalysts; Cell Culture Techniques; Diseases; Enzymes; Escherichia coli; Escherichia coli - metabolism; Fundamental and applied biological sciences. Psychology; Genetic engineering; Genetic technics; Hydrogen-Ion Concentration; Indican; Indican - metabolism; Indigo; Indirubin; Indoles - metabolism; Media; Methods. Procedures. Technologies; Non-recombinant; Optimization; Purity; Temperature; Indican; Indirubin; Indigo; Non-recombinant; Escherichia coli; Recombination; Bacteria; Genetically modified microorganism; Enterobacteriaceae
• ISSN: 0960-8524; 1873-2976
• DOI: 10.1016/j.biortech.2011.06.072

► We show that non-recombinant E. coli is able to produce indirubin from indican. ► The yield is reliably determined to be about 25-35%. ► Further increase in yield (1.8-2.1 folds) is achieved by re-adding whole cells. ► This process is more effective than the typical process using indole as a substrate. Indirubin is an important natural substance and has positive effects on various diseases. However, the current process of producing indirubin is inefficient, making it difficult to produce indirubin of high purity; thus, it is commercially unavailable. In this study, a method of indirubin using non-recombinant Escherichia coli as a whole cell enzyme with indican as a substrate was developed. After confirming that indirubin was produced from indican by non-recombinant E. coli under general conditions, attempts to compare the yield and purity of indirubin were conducted under various pH, temperature and culturing media conditions. Under the optimum conditions, the yield was reliably determined to be about 25–35%, and it was further increased (1.8–2.1 fold) by replenishing the catalyst with freshly prepared whole cells. Since the established method was simple and reproducible, high purity indirubin would expected to be produced efficiently through improvement of whole cell enzymes and development of scale-up processes.