Improved growth conditions for hyphomicrobium sp. B-522 and two additional strains
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Growth yields and rates of 3 hyphomicrobia were improved by varying components of or adding compounds to medium 337. Methanol (0.5% v/v), and similarly methylamine·HCl (3.38g/l), were optimal among 22 C-sources tested; increasing the methylamine·HCl concentration to 5.07g/l gave higher Hyphomicrobium B-522 yields but also prolonged lag periods. Ten C-sources (organic acids, alcohols) stimulated growth slightly but significantly, even in subcultures. Sugar compounds were not utilized. Strains B-522 and ZV-580 were stimulated by l-lysine and gluconate, while NQ-521 gr was stimulated by aspartate.
N-Sources tested were inorganic (3), organic (3), or complex (3). (NH4)2SO4 (0.5g/l) was optimal for strains ZV-580 and NQ-521 gr, but Hyphomicrobium B-522 grew best with urea-N. With NH 4 + , strain B-522 grew as homogeneous suspension, all other N-sources caused clumping and pellicle formation. Inorganic requirements (PO 4 3- , Mg, Ca, Fe, Mn, Mo) of strains B-522 and ZV-580 were optimized. Addition of Ni, Co, or Zn had no effect; metals “44” or Cu, resulted in growth inhibition.
Vitamin B12 stimulated Hyphomicrobium B-522; 2.5μg/l B12 decreased the doubling time from 9.3–10.8h to 5.4–5.8h. All combined single improvements resulted in a protein increase of 557% (B-522), 141% (NQ-521 gr), or 109% (ZV-580), respectively.
Key wordsHyphomicrobium Growth requirements Growth improvement Vitamin B12 C-1 utilization
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