여러가지 탄소원에 의한 Hansenula polymoypha의 Alcohol-oxidase합성

Synthesis lit Alcohol-oxidase in Hansenula polymoypha on Various Carbon Sources

Hansenula polymoypha의 배양조건과 배지의 탄소원에 따른 alcohol-oxidase의 생성능을 비교하기 위하여 H. polymorpha CBS 4732, H. polymorpha CBM 11, 그리고 H. polymorpha Cooney의 3균주에 대하여 실험하였다. 배양조건에 따른 효소생성은 glucose를 탄소원으로 첨가한 mineral salt medium에서 균체를 정상기까지 배양하여 methanol이 함유된 배지에서 재배양하는 2단계 배양법의 경우가 균체를 직접 methanol 배지에 배양한 경우보다 효율적이었으며 생성된 총효소량은 전자의 경우가 후자에 비해 1.9배정도 많았다. 다음 탄소원에 따른 효소생성능을 비교하였다. 먼저 직쇄 alcohol을 탄소원으로 사용한 경우 mothanol을 제외한 ethanol, propanol, butanoL, 그리고 pentanol에서는 효소가 생성되지 않았고 또한 이 직쇄 alcohol을 methanol과 혼합한 경우에도 극미량의 생성에 그쳤다. 여러 가지 탄소화합물 (glucose, xylose, lactose, glycerol, galactose, saccharose, sorbose, tactic acid acetic acid)을 탄소원으로 사용하면 methanol에 의해 생성된 효소량의 1/10 정도 생성되었고 이들을 methanol과 혼합사용하면 효소생성량은 급격히 증가하였고, 특히 lactose와 tactic acid의 경 우는 methanol 단독사용시 보다 오히려 다량 생성되었다. 시험 3균주 중에서는 어떤 경우에서든지 H. polymorpha CBS 4732가 alcohol-oxldase 생성능이 가장 좋은 것으로 나타났다.

The regulation of the synthesis of alcohol-oxidase (E.C.1.1.3.13.) was investigated in the methanol-utilizing yeasts during growth on different carbon sources. For this experiment, Hansenula polymoypha CBS 4132, Hansenula polymoypha CBM 11 and Hansenula polymoypha Cooney were cultured in mineral salt medium by changing its carbon sources. The production of alcohol-oxidase was varied by the carbon sources. For exmaple, alcohol-oxidase was undetectable: in all strains submitted to the test in the medium with glucose, but its production was rapidly increased when the carbon source was changed from glucose to methanol after 30 hrs of incubation. Moreover, this enzyme was not synthesized during growth on the primary aliphatic alcohols alone (ethanol, propanol, butanol or pentanol) or on the mixed substrates (0.5% methanol + 0.5% primary aliphatic alcohols). When cells were grown on the various carbon sources (glucose, xylose, lactose, glycerol, galactose, saccharose, sorbose, lactic acid or acetic acid), the alcohol-oxidase was about one-tenth of the activity found in cells grown on methanol alone. These carbon sources together with methanol yielded far better synthesis of alcohol-oxidase than in case of carbon sources alone. Especially, the alcohol-oxidase activity of the cells grown on lactose or lactic acid together with methanol was far better or similar than that of cells grown on methanol alone. The synthetic activity of alcohol-oxidase of Hansenula polymoypha CBS 4132 was the strongest among the three strains tested in every respect.