Use of soybean oil and ammonium sulfate additions to optimize secondary metabolite production.
A valine-overproducing mutant (MA7040, Streptomyces hygroscopicus) was found to produce 1.5 to 2.0 g/L of the immunoregulant, L-683,590, at the 0.6 m3 fermentation scale in a simple batch process using soybean oil and ammonium sulfate-based GYG5 medium. Levels of both lower (L-683,795) and higher (HH1 and HH2) undesirable homolog levels were controlled adequately. This batch process was utilized to produce broth economically at the 19 m3 fermentation scale. Material of acceptable purity was obtained without the multiple pure crystallizations previously required for an earlier culture, MA6678, requiring valine supplementation for impurity control. Investigations at the 0.6 m3 fermentation scale were conducted, varying agitation, pH, initial soybean oil/ammonium sulfate charges, and initial aeration rate to further improve growth and productivity. Mid-cycle ammonia levels and lipase activity appeared to have an important role. Using mid-cycle soybean oil additions, a titer of 2.3 g/L of L-683,590 was obtained, while titers reached 2.7 g/L using mid-cycle soybean oil and ammonium sulfate additions. Both higher and lower homolog levels remained acceptable during this fed-batch process. Optimal timing of mid-cycle oil and ammonium sulfate additions was considered a critical factor to further titer improvements.
Junker
B
B
Merck Research Laboratories, Building R810-120, Merck Research Laboratories, PO Box 2000, Rahway, New Jersey 07065, USA. beth_junker@merck.com
Mann
Z
Z
Gailliot
P
P
Byrne
K
K
Wilson
J
J
D000645:Ammonium Sulfate / Q000378:metabolism* / Q000494:pharmacology; D001709:Biotechnology / Q000379:methods; D002244:Carbon / Q000378:metabolism; D003470:Culture Media; D005285:Fermentation; D006863:Hydrogen-Ion Concentration; D008049:Lipase / Q000032:analysis; D009584:Nitrogen / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism; D000644:Quaternary Ammonium Compounds / Q000378:metabolism; D013024:Soybean Oil / Q000378:metabolism* / Q000494:pharmacology; D013302:Streptomyces / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D014633:Valine / Q000378:metabolism
D003470:Culture Media; D000644:Quaternary Ammonium Compounds; D002244:Carbon; D013024:Soybean Oil; D008049:Lipase; D014633:Valine; D009584:Nitrogen; D010100:Oxygen; D000645:Ammonium Sulfate